Explant culture skin.avi

Phasecontrast timelapse of a skin explant (piece of tissue). The explant sits in the lower right hand corner, the cells are growing out radially from the phase-bright edge of the tissue onto the free substrate. There are more and more cells by movement and division at the same time.




Mammalian cells can be grown over prolonged periods in tissue culture [http://en.wikipedia.org/wiki/Cell_culture] by repeated subculture, but how does one get the first set of cells? One possibilities is to digest the extracellular matrix away and sediment the cells liberated (i.e. by centrifugation). Another possibility is "explant culture" [http://en.wikipedia.org/wiki/Explant_culture] illustrated here: one takes a piece of tissue (here skin) chops it into very small pieces (smaller 1mm in any direction) and places the pieces into a dish or flask with a small amount of appropriate tissue culture medium (here L14, 10%FCS, antibiotics, glutamine) medium (0.5-1ml/25cm^2 flask). Cells have membrane bound matrix metalloproteases and also release such enzymes which digest away at the extracellular matrix, allowing the cells to crawl out of the piece of tissue onto the tissue culture substrate. These cells can later easily be digested away from the surface using trypsin, and can be maintained in cell culture. After a short period of repeated culture these cells will have lost crucial markers of differentiation though. Technicalities: Time lapse taken on a Zeiss Axiovert 25 using a 10x objective, a CCD camera, an Alrad imaging board and software at 1 frame every 2 mins, then this was reduced by a factor of 10 for viewing; the bar in the first image is 25micron.

Category:

Science & Technology

Tags:

• timelapse
• tissue culture
• skin
• explant
• biology

[jamesbush1994]

Great video, I've been inspired.