Resently，I try to culture primary intestinal epithelial cell in vitro. And I get some islets of cobblestone-like cells which looks like epithelial cell. However, this cells are hardly digested by 0.25%trpsin-EDTA and could not keep their morphology and survive after passage. I try various times but all fail. The method I used: I cover the tissue plate with collage type 1 and adapt cell medium with supplementation of 20ng/ml EGF, 10μg/ml insulin in DEME/F12.
By the way, I read an article which claimed that they successfully passaged the chicken primary intestinal epithelial cell for more than 50. That is amazing, but I do not see more article about this specially cell line DIV. That is amazing but weird, and I can not contact with those authors for learning more details..so sad
Dear Dahai Yu,
Primary cell culture is difficult always than cell line. First of all, yo can try only mechanical dissociation or combined with low concentration enzymatic method.
Ishak Ozel Tekin yes，now I really know the difficult in primary cell cultures. I suspended this experiment until I get more knowledge in intestinal cell differention and some skills in cell line construction. Maybe primary intestinal epithelial cells experient polarity in vitro so that I cannot continue culture this cell, I don't know about that, just guest.
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