Experimental study of isolation, culture and differentiation of islet stem cells in neonatal rat / 中国普通外科杂志

ABSTRACT

Objective To explore the method of isolation, identification and in vitro differentiation of islet stem cells in neonatal rat. Methods The whole pancreata of neonatal rats were digested with collagenase, then,under the (condition) of pH 7.4～7.6,the digested tissue fragments were cultured with serum and serum-free RPMI 1640 (( bFGF), EGF, N2).The whole formation process of new islet cell-like cell masses was examined.The insulin (release) test was used to detect islet function. The expression of nestin was tested by immunocytochemistry. Results The nestin positive cells can be found within 36 h of culture of the digested pancreatic fragments.After addition of bFGF,ECF,N2,nestin positive cells proliferated fast and formed new islets-like cell masses after 18～24 d, and (insulin)ssion could be confirmed. Conclusions The nestin positive cells of pancreatic cells possess the character of islet stem cells, and can form islet like cell masses through culture in vitro.