A method for the isolation and culture of human colonic crypts in collagen gels.
RH Whitehead, A Brown, PS Bhathal
In Vitro Cellular & Developmental Biology | Published : 1987
Little is known concerning the biological factors that control the proliferation of the stem cells of the colonic mucosa. In part this is due to a lack of systems suitable for studying the proliferation of this mucosa in vitro. We describe a simple technique for the isolation of single viable intact crypts which are free of stroma and which can then be cultured for periods of at least 16 d using a collagen gel culture method. This method of crypt isolation was efficient with the mean yield of viable intact crypts being 1.4 +/- 1.2 X 10(4) (mean +/- 1 SD) crypts/cm2 of mucosa. In culture, mucosal cells only survived for extended periods when the crypts were cultured in collagen gels over a fe..View full abstract