Yes. We have been making cultures routinely from 5-mm punch biopsies - is that comparably small to what you wanted? You can use the standard methods of trypsin or dispase splitting (we're using dispase now) plus dissociation of epidermis with trypsin-EDTA, and then grow the cells in a melanocyte medium on mouse keratinocyte feeders. (Mitomycin-treated). Even in the presence of SCF/EDN1/TPA/cholera toxin, the feeders still make a big difference for very small starting cell numbers. Once the melanocytes grow up to normal monolayer density, you don't need the feeders any more.
Methods for feeder cells and other related methods are here:
http://www.sgul.ac.uk/depts/anatomy/pages/dcbm&m.htm
Our laboratory establishes primary human cultures from foreskin samples. It is routine to use trypsin or dispase overnight following disinfection/PBS wash of the tissue and removal of the subcutaneous fat. Cut the tissue into small pieces and store in a small dish at 4C O/N. The next day, place the dish at 37C for 5 mins. In the meantime, prepare a flask with media containing growth factors for enrichment of melanocytes. Remove dish from 37C and strip off the epidermal layer from remaining tissue. Place epidermal pieces into 15ml conical containing the same melanocyte growth media used in the flask. Vortex the tube twice in 1 min intervals. Remove all media/tissue and place into flask containing media.
We don't use feeder cells. Typically we start out new cultures in T75 flasks, but you can also go smaller, since melanocytes tend to grow better when they are close to each other.
http://jcs.biologists.org/content/106/4/1323.long
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