 So welcome all to this SIB Virtual Computational Biology Seminar Series. Today we have the pleasure to host Amos Beihok, professor of bioinformatics and director of Department of Human Protein Sciences of the University of Geneva. And Amos is also the head of the CalIFO group. CalIFO stands for Computer and Laboratory Investigation of Proteins of Human Origin of the Swiss Institutes of Bioinformatics. I will try to summarize Amos' life in three minutes, which would probably be difficult. So I hope I'm not going to miss any main achievement in his career. So until June 2009, Amos was the head of SwissProt Group, which develops the SwissProt Knowledge Base as well as pro-site database of protein families and domains, as well as enzyme, a database of information of the nomenclature of enzymes. It was also responsible for the development of EXPASY, the world's first website dedicated to protein molecular biology. Amos Beihok's work mainly lies in the field of protein sequences analysis and the development of databases and software tools for this purpose. His most important contribution is the input of human knowledge by careful manual annotation in protein-related data. His first project as a PhD student was the development of a PC gene, an MS-DOS-based software package for the analysis of protein and nucleotide sequences. While working on this software package, he started to develop an annotated protein sequence database which became SwissProt, first released in July 1986. From 1988 onward, it has been a collaborative project with the data library group of the EMBL, which has now become the EBI, and SwissProt has grown both in size and in scope and in the amount of work and people necessary to produce it. Since 2003, the SwissProt team is part of the UniProt Consortium, and together with the EBI and the PIR, the Protein Information Resource, SwissProt produces the world's most comprehensive protein database, UniProt KB SwissProt as well as UniProt KB Tramble, UniREF and UniPark. In 1998, I was with some colleagues in Geneva and Lausanne and set up the Swiss Institute of Bioinformatics, known as the SIB, whose mission is to establish in Switzerland the center of excellence in the field of bioinformatics with an emphasis on research, education services and the development of databases and tools. Since 2009 in the framework of Califogroup, Amos is involved in the development of NexProt, a resource which aims to provide life scientists with a broad spectrum of knowledge on the human proteins. Today, Amos will tell us about another resource that the SIB has been developing, which is called CELOSORUS, a knowledge resource on cell lines which attempts to describe all cell lines used in biomedical research. Amos, thanks again for accepting our invitation, and the floor is yours. Thank you very much, Diana, for this nice introduction. And as you say, in fact, I will speak about something which has nothing to do with my work and all my career on protein, a new passion of mine on cell lines. So there will be discussing bioinformatics resource on cell lines and also things which have nothing to do with bioinformatics. In fact, most of my talk has nothing to do with bioinformatics. Part of it is to warn people about cell line contamination. And the biggest part of the talk are human interest stories behind cell lines. So it will be more sociological than, in fact, bioinformatics or science. Okay, let's start with cell lines. I mean, they're used all over the world by labs, academics labs, industrial labs. So, I mean, basically, most labs doing research one day or another are using cell lines. And people acquire them either through cell line collection, most well-known ones are ATCC, DSMZ, and so on, or more often than, I mean, by buying them, people get them from the lab next door. You know, a postdoc goes and says to the next postdoc, to the next lab, can you give me, I mean, this cell line I needed for my research, and so on. So a lot of cell lines get, in fact, sent from lab to lab, from postdoc to postdoc, from main scientist to other scientists. Anyway, cell lines are everywhere. And so there's a lot of different type of cell lines. Of course, there are also cancer cell lines, which many people know with ELA and others, but there are also a lot of transformed cell line by different, I mean, methods. You can transform cell line with virus, with radiation, with mutagenesis and so on, and of course embryonic stem cell and pre-potent cells, and of course also hybridomas, which are used to, in fact, make monoclonal antibodies. Anyway, a lot of different type of cell lines. And in terms of resource, what did you have until the cell service came out? Well, you have cell line catalogs. All of the cell line collection generally offers a cell line through online catalogs. You have a number of ontology, which I will describe, and many specialized database, which either are based on cell line or report results of experiments and are linked, I mean, are linking those experiments to cell line, like in code, cosmic, and others. Anyway, up to now, there was no single resource where all this information is available. So let's start with a little bit with bioinformatics and let's start with the source of a lot of the data, which is the cell line collection. Now, there is about almost 50 different entities which distribute cell lines all over the world, I mean, in Europe, US, and well, most continent except Africa. And those groups are not at all, I mean, savvy in terms of bioinformatics. They know how to ship cell lines, they know how to conserve them, they know how to distribute them, but they don't know how to basically make use of the information they have collected and they don't know how to put it on the web. So, I mean, something like a year and a half ago, I drafted five minimal requirements, which are very basic. I mean, each cell collection should have at least one individual page for each cell line. That's a page URL should be based on the catalogue number so that you can, knowing the catalogue, go directly to that page. And it will be useful to have a mechanism to know what are the new cell lines and what are the ones which are discontinuated because this is a work in progress. Cell line collection, every month come up with new cell line and say discontinues some cell line for different type of reasons, one of it being cell line contamination. And also, it's availability of STR profile. Here, I won't explain what STR profile, I will do so in a few slides, but anyway it's an important item of information which allows to know which cell line is which, especially for human cell line. Now, the current situation is the following. This is just a spreadsheet. In green are compliance. You have five columns. If you can see, there is zero, I mean, green compliance over the five easy rules to basically to follow. And six even have zero compliance. Basically, they're completely red. I mean, they don't have individual page. They don't have a way of getting to their catalogue and so on. So it's changing very slowly, but it's changing a bit. And hopefully, I mean, in one or two years, there will be a little bit more green here. But you see, it's a big problem because those groups have basically no knowledge of what it means to have unique identifier, stable identifier. They change their catalogue. They would change the URL of their page. And the same cell line suddenly is found somewhere else and you don't even know if it's the same one or not. Anyway, let's go to things which are in fact more stable and which are as bioinformatician use. Those are ontology. When I started the Salazarus idea was not to create a new resource but to use existing resource. So what was there which existed? Main one is the cell line ontology, which is a wonderful resource, almost 40,000 terms, 30,000 cell line terms. Unfortunately, in fact, it has a high degree of redundancy in the cell line terms because it creates as many, I mean, entries for cell line as a distributor of the same cell line. So that's a little bit problematic. But it's of high quality in terms of the definition of cell type and cell categories. So as an ontology of cell line types, it's perfect. As a dictionary or a Tesorius of cell lines, it's neither complete nor, I mean, and it's basically redundant. Now, there was another cell line ontology done in India which, I mean, stopped. I mean, it's no longer maintained. It's no longer a connection cell line ontology. Unfortunately, not maintained anymore. And then you have Brenda, which does a very good job of building an ontology on every tissue, strain, and cells that they use in Brenda. And so I have 2,000 cell lines in the BTO, but it's only the ones that they use inside Brenda. So, yes, it's nice, but it's not really, I mean, complete. You have also EBI creating experimental factor ontology, which has 1,300 cell line. Again, it's a cell line which I use by some resource at EBI and also in AIN code. You have the beta cell genomic ontology, only a few cell lines, and even mesh, which has 24 cell lines in it. So that's for cell line ontology. Basically, there are resources. Each of them is okay, but it's nowhere complete and for what we needed to do far from being what we needed. Okay. Before we go to the cellosaurus, I will make a little bit of detour through a number of other problems with cell line. One of it is naming issues. I mean, short cell line names are a disaster. Here you have 10 different cell line names, all of it two or three characters. And in fact, those names are used by 37 different cell lines. So each of them here has at least three or four different cell line associated. And PC3 can be a prostate carcinoma cell line, which is very well known, but it's also a lung cancer cell line. It's also a mouse cell line and so on. And so the problem is even when people propose longer names, well, then authors try to abbreviate them. So you have a cell line called FG2C3A, a derivative of FG2. That's okay to call it like this, but then people will only call it C3A. And of course C3A is not unique. So that's a problem. So there are a few nomenclature rules, not really, I mean, a lot. There was one for insect cell line, which is quite well followed, and one for embryonic stem cell and iPSC, which is not yet well enforced. And of course, in addition to that, the big problem is that in the literature you have a huge number of misspelling of the cell line. It's quite a mess to find in paper what cell line have been used. Okay, let's get to the second problem with cell line, which is the biggest one. It's a cell line contamination issue. And these are just titles of papers or, I mean, blogs. I mean, on the last three or four years. I mean, basically, where people, I mean, are reporting about this problem. A lot of cell line, especially in tumor field because it's especially true of cancer cell line, get contaminated. And in fact, estimate of the range of cancer, extent of cancer cell line contamination range from 20 to 36%. This is quite enormous. It means that one cell line out of four in a paper is not what the person think it is. So that's, I mean, quite, I mean, frightening. And a lot of people are trying to help with this. One of those effort is ICLAC, the International Cell Line Authentification Committee, which was set up, I mean, a number of years ago by Amanda Cape-Davis, and which has members of different cell line collections since it's working on cell line. It tries to give good practice, I mean, information also on how to name cell line, but also what's important, it has, in fact, a list of all known contaminated cell line. Inside, I mean, if you click on it in resource, you will find it. Now, I mean, that's, I mean, quite an effort, and they try to spread the word around in conference, in different place, this problem of cell line contamination. But it's, I mean, the best way to get rid of this problem is to test cell line, to authenticate cell. And that's where those STR profile, I mean, comes up, which I mentioned just a few minutes ago. So what are STR profile? Short tandem repeats. Those are loci in the human genomes, which are both repeated, short, short bits of DNA, and quite polymorphic. And of course, you know about them, because those are the ones which are used in forensic identification and paternity testing. So every good American TV series, as at one point or another, somebody is going to do some DNA identification using those STR. Now, those STR can also be used to ensure the quality and integrity of human cell lines. And there is an NC standards that make use of 18 markers. So 18 of those short STR, while one is not really an STR, amelogenin, it's just because amelogenin gene exists in two copies, one on chromosome X and one on chromosome Y. So this marker is used to make sure if the gender of the cell line is correct. With some caveats, many male cell lines can lose their Y chromosome. So people have also thought, oh, this is not the right cell line. I'm supposed to have a male cell line and I got only X as amelogenin. It's not true. But the reverse is true. If you think you have a female cell line and you have a Y, you're in trouble. So this is for human. It's an initiative to establish both panels of STR markers for males and dog cell lines. So this is going on. And what should people do? Well, they should basically send a cell line to companies. There's a lot of companies, including cell collection, where basically you send your cell line, the report, I mean to give you back a report with a different allele that you can find for each of the different STR markers, like here you have for CSF1PO, this cell line as a hit on 10 and 11 copies of this repeat. And so basically then you get a report saying your cell line is identical as an STR profile identical to the cell line XYZ. And if it was really XYZ, you were using UAPI. Or, I mean, you get a bad news saying your cell line does not have this profile. It has a profile of another one. One of the problems with this, those companies do that job very well. But unfortunately, most of them try to keep the database of STR profiles that they use as a private thing because the more profiles they have, the more they can test, and they don't want to send it to other competitors. Which beats a little bit, I mean, the principle of having the STR profile available to all so that people can compare them. Okay, interest of cell others. So as Jenna said, it's a knowledge resource on cell line. And it has a scope which covers all kinds of immortalized cell line. And including natural immortal cell line like stem cell line, embryonic stem cell line. And also some fine act life cell line when those are distributed and used by widely. It's scope in terms of species, it's vertebrate and invertebrate in second ticks mainly, but not plant cell lines and not primary cells which are not cell lines. Those are cell culture and so on. So it basically includes everything which is known as cell line except for plant cell lines. Now what do you find in cell others? You find a lot of things. I mean, I put it on the slide in tree slide, but I'm not going to go through all of it. Of course, you have the cell name and synonyms. That was the beginning of cellosaurus. The cellosaurus started out as cell tesoris, therefore, cellosaurus. But it became, I mean, a knowledge base because it became much more than just cell line names and synonyms and misspellings. But of course, the category, is it a cancer cell line or transform and so on. The sex of the individual from which it was established which is very important. The species of origin, of course, I mean, it's not only human cell line, we will see a lot of different species for non-human cell line, whether it's a subspecies or breed of the animal, known contamination and miscertification. So all of the information from ICLAC is integrated into cellosaurus. So every known contaminated and miscertified cell line is there. Annotation of the disease of disease cell line, meaning cancer and genetic disease cell line, using the NCI tesoris, and a lot of other things. Genes are transfected, genes which are knocked out or edited. If a cell line has been selected by resistance to a compound, a lot of cell line are tested, are challenged with, for example, chemotherapy drugs and so on. And then, I mean, people then develop new cell line which are resistant to those drugs, to use them for research. So transformers used to immortalize a cell line, monoclonal antibody targets, and a lot of other things. I mean, so population doubling time and so on. And getting to the end of that list, an important thing, so STR markers. And basically this is collected from cell line collection by asking them, nagging them, also publication, all of the publication where cell lines, STR markers have been published and a lot of personal communication. Basically I go and ask everyone which says that they have their cell line authenticated to send this information, and a lot of people do. Now, it's a manually created database and links to a huge number of resources because there is also specialized resources that make use of cell line, and of course, all of the cell line collection, you see ATCC, DSMZ, Coriel, which is the biggest distributor of cell line in the world, about 20,000 different cell line, mainly for disease, genetic disease. I mean, it's a huge institution, Coriel, like ATCC, maybe less well known for people which are not working on genetic disease. And so where do we stand now with cellosaurus? 83,000 cell line from 564 species, but 90% are from three species, human, mouse, and rat. And 40,000 synonyms, almost, well, more than 60,000 reference to 13,400 publication, and a lot of cross-reference to all of those resources I was showing on the slide before. 10,000 web links, and now there are 5,000 human cell line with STR data. Now, where can you get it? You can browse on Xpazee, where I mean, you just, I mean, Google cellosaurus and you will learn on the page on Xpazee. And of course, as by information, you may want to use it in another resource, so you can FTP it, and so it's available in three different formats, a structure flat file, which look a lot like, guess what, a swiss plot format? I mean, you may wonder why. And a noble format file, and what is more interesting for people working in bioinformatics, an XML file. And I would, I mean, tell people which are serious in using it to use the XML version of it. Of course. Now, cellosaurus usage on Xpazee, I mean, it has been put on June 2015, so two years ago. And for the first two months, it was totally private. I mean, it was the first part of those hits or just local hits. And then it became available, and as you see, the use is growing quite well. You see maybe two dips. You can see that Christmas has an influence on usage of resource, I mean, in the world. And so those are those two big dips. And what's interesting is we can still so very important for scientists, I mean. And you can see those dip every week. And what's very pleasing for us is that 40% of the user are returning visitors, so that people which come to cellaros come back again, which is, and this is, those numbers are growing. And I looked at it for the last month, it's more like 50% now of people are returning users. One important thing still in terms of bioinformatics is resource identification initiatives. Now, this is an initiative which started, I think, two years ago, three years ago in U.S., which introduced the concept of resource, research resource identifier, error IDs. And so the idea is that you should have a persistent and unique identifier for every resource that you use in experiment. Antibody, cell line, organism, and tools, software tools also, I mean. And so basically they started this effort to try to push publishers to have authors put those error IDs, and of course they needed resource which can be cited and force the cell lines to use the cellosaurus as a cell line resource. And basically what you have to do in papers is put this error ID followed by a tag like AB for antibody registry, CVCL, which is a prefix of the cellosaurus accession number for cellosaurus. And basically this will allow, I mean, retrieval of the information, automatic retrieval and tracing back which is a resource which was used. And there is a portal. Oops, there is a resource certification issue. You will have the URL of the portal which just came down, is missing from this slide. So here is an example of, I mean, a part of a method of a paper which was published a few weeks ago in Elife and overlined in red. You see in the section of cell lines that they have indicated both in text. So raw 264 cell line comes from there and put it, you know, which is their error ID. And also those people have done a good job because they have authenticated their cell line by stair profiling and verified that they were also negative for mycoplasma. That's a good thing. Okay, three slides to finish a bioinformatic part which is just screenshot to show you a little bit of the cellosaurus look like on Xpasi when you browse to it. You have the name of cell line, you have information in red when it's problem-adminated. You have publication. You have links to the species, to the disease, to the parent cell line. You can have web links. You can have information on what has been done on this cell line. For example, deep proteome analysis that it's part of two more cell panels that it's used by INCODE and so on. So you have a lot of information like this. And, of course, the STR profile. And here you see the markers and you see the values for the different allele. And what you can see also is that, I mean, some cell line can be verified by different people which find sometimes different results. And this is normal because cell line is a living organism and they can lose copies of chromosomes or they can even acquire copies of chromosomes. And, for example, this cell line on chromosome 8 on the marker D8S1179, as you can see, has three different allele because normally it should be two or one. I mean, if it's a mosaic of that position, it has three because probably it has a triplication of that part of chromosome 8. And here you see that on this allele is a result but which more or less corroborate and that it just as some of them have missed the allele of 11 copies on one chromosome and the other one has missed the one of 13 copies and so on. Anyway, STR profile and, of course, cross-reference to a lot of resource. So that's the end. Oh, yes, sorry. No, it's not the end. I still have one slide is what can be added cellosterous in terms of tool to help people to compare STR profile. So, I mean, bone-formatic tools where people would interest the STR profile and compare it to what there is. And also, I mean, I'm working closely with the people from Wikidata in terms of integration of part of the cellosterous data into Wikidata so that it be part of the semantic web. But this, if people are interested we can discuss it as a question time. In terms of content, some people are asking for to integrate a non-contamination derived xenograft which are not really cell-lined but close to cell-lined. And, of course, a lot of other things can be added like the tissue of origin which is not yet there, info regarding the age of patient or animal, the virus which are integrated in cell lines, the translocation for cancer cell line and bio-safety level industries asking that this information be put in. Okay. Now, as I said, that's end of bone-formatics and contamination which is what is there behind those cell lines? Of course, all of you know Hila. And most of you, I guess, know about Hila stories. It was established in 1951. It was the first immortal human cell line established at John Hopkins by Zalab and by Sir Fort of George K. and his wife. And it's the most well-known and widely used cancer cell line. Now, it was obtained from a medical tumor of a patient known as Henrietta K. which was a 31-year-old patient that died in October of 1951. Here is a picture, a nice picture of her before she was sick. And, in fact, this story is well-known because, in fact, it has been described in an incredible book which is this book, The Immortal Life on Henrietta Lacks which I would consult to anyone to read. It's really like a detective story telling you all about the history of this cell line or it got well-known and it's also the history of this Lacks family. Now, the big problem with this cell line is there is a lack of consent from the family of Henrietta Lacks and by herself that her tissue would be used to make a cell line. And some people have said this is all, it was just used in 1950 when you went to an hospital doctors could do whatever they wanted on your samples and nobody would have even thought of asking a patient if they were giving permission for the sample. So, in fact, she didn't know that her cell line was made out of her tissue. Well, she died anyway quite quickly after the tissue was used but her family never heard of it until in the late 80s. And it became a big issue because in 2013 the genome of Hila was sequenced by different groups and the family was not really happy about the disclosure and in fact, I mean, in some place if you want to have access not to the Hila genome, you need to have permission. Some others have put part of the Hila genome. It's almost like, you know, closing the door of the barn after the animal has got out. I mean, part of the genome of Hila everywhere. It has been sequenced so many times that there's nothing the family can do, unfortunately, but they're not very happy about it. So, it got to an agreement with NIH about such genomic data could be used if people ask for permission. Since then, they are suing now John Hopkins. I mean, because in fact what happens, it was also a very nice movie two months ago on Hila, which came out in US and it renewed the interest on Hila and thought, well, maybe we should sue John Hopkins. So, it's a story which, I mean, is still evolving. We don't know what's going to happen in terms of the Hila, Lax family and Hila. Now, this is a story Hila which is well known, but there's a lot of other stories which are less well known. Here's another story. Of course, Hila is a very aggressive cell line and it's the one which is in fact contaminating a lot of other cell line. And for the very beginning of the 1960, I mean, from the end of 50 to the 70s, it was contaminating so many cell lines that basically a lot of the research which was done on cell line which was supposed to be from normal human tissue was done, in fact, on cancer cell line. And one of those cancers, I mean, normal cell line which is not normal but Hila is wish. Now wish was established by someone which is very well known, Leonardo Affleck. And Leonardo Affleck is an American scientist which discovered in 1960 that in fact normal cell line are not supposed to be immortal and that after a number of duplications they die. This became known as the Affleck limit and of course it took 30 to 40 years to understand that this was due to the shortening of the telomerase. But much before it was known that normal cell line would die. But at the time he didn't know about it so he was trained to establish a cell line. So what did he do? He tried to use amniotic sac in which his daughter was born and this is what he published in conformity with the recommendation made by committees set up to establish rule for naming cell lines. A new cell line has been named wish for Wister Institute, Susan Affleck. So basically the privacy of or the consent of his daughter was not really asked and she was. But I mean this doesn't really matter because it's not anyway the genome of Susan Affleck which is in the wish cell line it's Hila. But you see that at the time I mean people were proud of not only of things that have established a cell line and where it came from they would give the full name and were of the individual where the cell line came from. Now there is another story about Affleck which is much in fact more complex and this is a story of a cell line called WRE38. So it's a finite length embryonic lung fibrous cell line. So it's a cell line which after a number of population doubling will die out like any normal fibroblast cell line. Now why did he create a cell line axis? It was at a time where people were trying to establish the first vaccine against polio, against rubella and other types in cell lines. And for polio they were using monkey cell lines. Unfortunately a lot of people found out that those monkey cell lines were infected with SV40 and it became quite scary because what could happen maybe people could get tumors from them and so on. So Leonhard Affleck and others tried to establish cell lines from human and which should be clean and he said if they took cell line from fetus those fetus would not be sick with any disease, would not have any virus and those could be used to grow vaccine. So he developed a number of cell line and each of them had a number. He started with WI1 and you could think this is number 38 in Syria in fact he went to 27 and came to 38 and nobody knows why. So it was a 28th cell line established from a female fetus. No, this cell line is very famous. I mean first reason is that it's a cell line which was used to produce the first rubella virus of vaccine sorry and it's still being used for the rubella vaccine. So in fact no, the rubella vaccine is part of a tetra or penta vaccine with different other, I mean the vaccine but it's a rubella part is still made using this cell line. And this is told in a story, a book from a Meredith Wadman which is a journalist in science and that book is also quite nice. It just came out six months ago and it's a detective story about vaccine or this fight on vaccine took 30 years to I mean grow and basically everything which happened with this race in establishing the rubella vaccine but other vaccine as well. So that's the first thing why this cell line is interesting. Second reason is that of course I mean he needed a fetus to I mean create this cell line. At the time in US of course abortion was not legal but you could get some fetus for medical I mean where there were medical abortion but still it was not very easy to get. So Leonard Affleck had a collaboration with Swedish I mean doctors and basically fetus which was used to produce a cell line was from a woman which had a number of children. She was as it says here married to an alcoholic and she didn't want another baby so she got an abortion which was legal in Sweden but difficult to get so she got it when she was four month pregnant and she didn't know that her fetus was used to derive a cell line. Of course nobody asked her at the time but she knew after some 10 or 15 years later what happened is when companies started selling the rubella vaccine they wanted to make sure that there was no I mean genetic problems with the cell line and of course you couldn't sequence at the time so they asked back to the Swedish doctors saying can we have the record of this woman to know if I mean there is no mental problem or no genetic problem in her family and so on and they went back to her and she was quite angry about it and in fact when one man contacted her in 2013 she basically told her that this was not something which should have been done, that this was all done with her permission and she is quite well angry and rightly angry and in fact we don't know who she is and that's I mean quite normal because one man which contacted her promised that her name would not be known and kept secret but you have somewhere in Sweden a very angry woman and she's rightly angry because all of this has been done with her permission. So that's not all for WI-38 so it's used as I said for vaccine with also another I mean finite cell line from her features in US from the Medical Research Council MRC-5 and it's a big issue for every right wing right lifers in US in other countries which says that I mean vaccine should not be built out of what they call baby parts so baby are not spare parts so this says that because those cell lines come from a fetus there is part of a fetus which was aborted inside the vaccine so children should not be vaccinated with such I mean a vaccine made out of fetal cell line so it went to the Vatican also and which made a report I mean the Pontifika Academia I mean which was led at the time by Cardinal Ratzinger which became Pope after and he basically said yes it's okay to vaccinate with those vaccine but he urged people to make life difficult for the pharmaceutical industry so since then our fight all over every time some companies come up with I mean so yearly I mean board meetings you have people with placards saying you know stop making a vaccine out of dead baby and so on so companies are moving out of making vaccine with human cell lines are using very old for monkeys are using chicken cell lines and so on and not I mean so pro-life first I've started being verlin not against the U.S. but against China because in fact the Chinese I've created a cell line called valvax and they want to use for vaccine and of course you have things like scientists in China create new or nine apported babies I mean okay so you see or WI-38 still as I mean is a cell line which is as a story now there is a four part of this story so Affleck developed WI-38 at the Vistar Institute where he was working with the NIH grant but he got into a fight with the director of Vistar and he moved to California okay stand forward but he took with him a fridge with I mean liquid nitrogen and he drove all over the U.S. with all of the little vials of WI-38 and then nobody really complained because what happened is after he would whenever somebody wanted this cell line it would ship at the cut of the cell line and he would just ask for a nominal fee for shipping so no more procedures so nobody cared but in 1974 he thought he should create a company he created a company called Cell Associates and then he went to companies producing vaccine and asked them for licensing fee now NIH suddenly started to be interested and so started an investigation which basically destroyed the career of Affleck he had to resign from where he was he was not in Stanford anymore but I don't remember where he was but anyway he had to resign and it took seven years of legitimization and it ended with the end of court I mean a decision and basically I mean for a long time his career was destroyed and it's only now that he's doing a lot of interviews he's 88 and he's seen by a lot of people as a precursor of the relationship between biotech industry and scientists at universities because those rules were making patterns from products which at a university started in 1980s at Stanford it's the first patterns for genetic production of insulin and so on but in 1974 it was seen more as something that a scientist should never do is create a company and make money out of it it was seen as he was evil now he's seen more as a precursor of things which have happened since okay another story MSCF7 most well known breast cancer saline 1971 stands for Michigan Cancer Foundation 7th try so they try with 6 other patients in work and they manage with 7-1 and the 7-1 we know is a donor because she's she was sorry a sister in a convent in Michigan so Ellen Marion was known as sister Catherine Francis and she had cancer in 1962 they tried to cure her cancer unfortunately it didn't work out and when she was dying her doctor asked if he could use her saline and she was happy to do so and the congregation was not only happy to allow this but also to publicize the fact that those salines come from her so this is a case where in fact the donor accepted and in the website of the congregation you will find the story of this nun and the doctor said that maybe one of the reasons that he managed to grow those salines and not the others before is because she prayed every day I'll let you decide if it's true or not now another cancer saline was developed in the late 70s by a group in California by Dr. David Golden and what happened was that they filled for a patent so that was a patent on a saline which was quite rare at the time to patent a saline but the problem is John Moore the guy from which the saline was established didn't know about it so he didn't know that there was a saline and he didn't know that it was patented so when he learned about it he sued the University of California and basically it was a big discussion but basically he could not get money from the University of California because he had consented to allow Gold to perform research on sample of his spleen after surgery and so part of this research could be a saline and once you had a saline you could patent it but what was very important is that Gold was in fact hit a little bit on the fingers by a judge which said that he had breached his fiduciary duty he should have informed John Moore of what was done with his samples so John Moore could not have money from this saline but he should have been informed at every step of what was done now what's interesting is who is this guy John Moore so he died in 2001 and you can see that this guy has a really interesting life because he was a surveying engineer on an Alaska old pipeline an alcolyse conceler a photographer a farm, I don't know if anybody is running a warm farm but anyway he sold seafood he was a sensor and beer distributor and then he was in saline marketing for an internet company so really it's somebody which had a lot of different skills and I think it's quite a very American life story of self-made men which went from one job to another one and his daughter I mean said that he was very adamant that the fact that patients should know what's happening to their body or to their body part now another story which is old but which is also interesting is the Kenyan Burkitt Saline so in 1947 an Hungarian scientist left Hungary because that was a time where it became a communist country he fled from Hungary went to Sweden and with his wife they opened a tumor biology center by institute and they worked on the connection between EBV and Burkitt lymphoma EBV had been discovered I mean a number of years before Burkitt lymphoma also but suddenly it became known that EBV could be a cause of Burkitt lymphoma so they used biopsies that were sent from hospitals in Kenya to establish salines from Burkitt lymphoma and a lot of you which I work on saline will know about DODI it's used everywhere as a type of Burkitt lymphoma saline it's an example of a saline which is used everywhere for studying Burkitt for Burkitt lymphoma but the thing is what they did is they named the saline with the first name of the patient that's already discontentable I mean it would be not this but what's worse is that the papers mentioned the last names so here I have I mean taken away the last names in fact just to tell you a little story when I started saliseries I put you know this saline comes from the cells of a patient named DODI such and such and a scientist I mean Amanda Cape Davis which run ICLAC told me you shouldn't put this those people have not given permission for it and I said yes I can find it on you know by just googling and finding those paper and she said yes but don't spread more of this information that what it should be as those people never gave their permission we should not put their names in any public database or resource and so on that's why you see here some black I mean box and it shows I mean that at the time people didn't care of saying the full name cities they lived and put the pictures and sometimes you have even pictures of the family and so on so that was deemed quite normal at the time what time is it ok another a few other short stories much shorter on I mean a number of other I mean saline so you see this saline with the paper that's the title of the paper I mean from groups in Germany you see that the first author doesn't have an institute on it it just says Apparote Rotenbach and if you go at the end of the paper it says that basically the first author is the person from whom the saline was established so basically that's only known I think paper I could find which describe a saline and I mean added the name of the person that's an author and even the first author on the paper so that's I mean quite I mean an interesting story now here are two small stories which are quite sad and then I will go to less sad stories it's about pediatric saline so unfortunately cancer in small children this is a saline from a neuroblastoma which was established in 2006 so neuroblastoma NB in 2006 from a small girl which died when she was nine and her father asked to name the saline and he was quite angry with I mean the disease I mean which killed his daughter and so he basically names the saline and such saline you could guess what FU stands for and basically I mean he wrote in a blog I mean it's probably not something that will be published in any scientific journal the reason for this saline is called FU NB 2006 now another sad story but which also shows more what people can do once they get out of this horrible trauma of losing a child it's this small girl Claire Vetsel McKenna which died seven year old from another very rare cancer diffuse intrinsic pointing glioma and what parents did, they created a foundation to help I mean studying this disease and basically I mean to try to I mean advance on this research on this cancer which is quite rare and so really in fact very active they raised over one million dollars over the last five years and basically I contacted him and I said I mean is it okay that these salines come from your daughter and so I point to your site and to your foundation and he says yes and you can tell the story of the foundation because the more people know about the foundation and if they are going to you know give money for research for this cancer the better it is so this is an example where in fact it's a private issue is less important for a family then making sure that this disease can be fought I mean in the most efficient way now yes there is still another story which sad story sorry I forgot about that one sorry still one sad story before going to more this is also a rare disease on it's nose cancer synonezol and different side carcinoma and I mean one of the person which had was diagnosed with this cancer was at the time a young medical student first year medical student in Tulane University in New Orleans and basically when he knew about it and knew that this cancer is fatal in 98 to 99% of the case he had to decide do he continue his study or do something completely for the next few years before he dies and in fact he decided that he wanted to continue his study but also try to do something to fight against this cancer so he convinced a very well known a metallurgist tiller curial to work in his lab to establish a cell line here is a nice drawing of tiller and while doing his studies so he managed to pass every year his exam doing all his radiation treatments he worked during the night in a lab it was really frustrating because he did manage first and he did manage to get a cell culture in 2004 he needed money to get this running so curial which is an ultra mountain racer what he did is basically he created something called the Bones for Life and he set up a world record by taking a basketball and running and dribbling a basketball for 108 miles for 24 so that's quite an achievement so he's in a Guinness book of records this guy for having done this to get money for this cell line unfortunately Andy Martin died in 2004 but the story continues is that in 2005 Hurricane Katrina destroyed most of the infrastructure research infrastructure of Tulane University including the labs where this cell line was but curial managed to save his cell line and he published a newspaper article where he basically managed to get it now you could say where is this cell line? now that's where the story is not finished I tried to contact Curielli, he doesn't want to answer me I'm going to nag him until I get to know what happened to this cell line because it was never published so last thing you can find is in 2010 seven years ago it was said that he was working on it but I mean I don't know what's happening with this cell line does it exist, was it lost or not but it's an interesting story okay so not tragic stories here is two examples and I will go fast on it one is a melanoma cell line established by a group of Ruth Alaban and she's a very well known Yale University melanoma researcher, she developed a number of melanoma cell line one of it from a patient whose name is known because he wrote a book called Cancer Gifts and he basically explained his fight against melanoma and basically he says for example I was forever proud to see UF in scientific presentation of melanoma researcher it stood for my cell line, U for Yale University and FHEF for FONAN and when Yale legal official told me UF was a too descriptive identifier and said it had to be changed to something less decipherable give me a break basically he's proud of it to change the cell name he basically makes it known that it is his cell line so it's an interesting guy because he studied journalism and then he worked for U.S. Senate and in 1970 he was one of the guy which managed to push President Carter to science the creation of the U.S. Department of Education I thought the Department of Education was something a ministry which existed for years and years in U.S. but no separate ministers there was no secretary of state for education so Carter signed the law to get and he was one of the person fighting for it then he worked in the context of article control trade he was director of those trade association and in 2007 for the next three years he had this horrible fight against melanoma radiation surgery interleaking treatment interferon and he was one of the first people to be saved by experimental adaptive T-cell therapy so in fact he was lucky to be one of the first and now together with his husband he basically stopped working in the field of article control trade and so there is journalists and they have a bed and breakfast in in Vermont and the white horse is in and you all welcome to go there so basically I contacted him and asked him about his story and he was very happy if we make some advertisement for his bed and breakfast in Vermont last story about cancer saline the John Hopkins Cordoma Line Cordoma is also a very rare cancer at the skull base or spine and GS C7 was one of the first lines of the disease established by a group of Alfredo Kinan's and is a famous brain surgeon and is an example of the stupidity of well there is a lot of stupidity in Trump policy but of immigration policy because you can see that in 87 he basically was one of the people which jumped the border got caught, got sent back to Mexico tried again and basically it's a type of success story an American success story he basically didn't know any word of English had no money but he managed to rise up in the culturally and being a doctor and a very well known neurosurgeon so his group established a saline and it's from a woman called Susan Garbet she is I mean Neto Baltimore, she's a retard kindergarten teacher in Florida she started writing her first book on Alzheimer's because her father had Alzheimer's so she wrote a book and then after a fight against Cordoma she wrote another book which is called Confronting Cordoma Cancer and she has also on Facebook she explains she went to the lab and looked at her saline and so on so basically because of that I could put two and two together I know that it was her saline and she shares a story and that it be known that GS7 is from her Cordoma cells last two slides nothing to do with cancers this is induced propratine stem cell and it's from Coriel's I mean libraries of salines they have also saline with GM and a number and it comes from someone which some of you may be heard of a guy called Craig Venter and he's so publicity shy that he announced it on the Craig Venter Institute that there was a saline an apparently healthy IPSC saline which was available from his fibroblast so obviously he doesn't want it to be I mean secrets that this saline is from his fibroblast and another guy which is not shy either was born from addition and which has two saline developed from his tissue this is George Church who is a very well known US geneticist was active in human genome E. coli genome is not trying to recreate a woolly mammoth and to synthesize a human genome I mean basically you hear about him every I would say month with another thing and he created something called the personal genome project that some of you know where people basically put their information on their genome and even like him their health record so of course you can find his genome and salines from his genome and here is another cryptic saline the last one and this is not a human one it just to say that it's not only human salines which can be established from individuals which we know who they are Clint is Chimp which was sequence and which was on the first page of Nature a number of years ago unfortunately it died just after this genome was published but at that time to create salines so Clint is dead long-lived Clint and so our fourth saline which has been established from his tissue so I don't think anybody asked for his constant but anyway I mean maybe he did we will never know about it so that is end of those human interest story at that I want to thank for all of this also Elizabeth Gasthiger because she implemented Salazaris on Xpazee Anagato in both in Geneva working Elizabeth working for the web team of Cecibe and Anagato for the California group and was developed a tool which allows the oboe and XML version to be generated from the flat file but also to thank 500 more than 500 scientists which answer nagging emails to get information on their salines Amanda Cape Davis which I said which is Secretary General of ICLAC she had a lot of suggestions for Salazaris and she led me to a number of those stories and those people which I mean agreed that their story be told here Susan Garbet, Robert Phonan and David Vetzel's father of Christine Vetzel so with this this is the end and I thank you for your attention thank you