Comparison of the effects on differentiation of mouse embryonic stem cells into insulin-secreting cells among three cell culture protocols
- VernacularTitle:三种不同细胞培养方式对体外小鼠胚胎干细胞分化为胰岛素分泌细胞诱导效果的比较
- Author:
Muchao WU
;
Hua CHENG
;
Lihong CHEN
;
Mingtong XU
;
Feng LI
;
Shengneng XUE
- Publication Type:Journal Article
- Keywords:
Embryonic stem cells;
Cell culture;
Insulin-secreting cells
- From:
Chinese Journal of Pathophysiology
2000;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To compare the effects of three different cell culture protocols: embryonic body(EB) formation,EB formation-monolayer and monolayer on differentiation of mouse embryonic stem(ES) cells into insulin-secreting cells.METHODS: E14.1 mouse ES cells were treated with GLP-1,betacellulin,activin A,bFGF and nicotinamide by using EB formation,EB formation-monolayer and monolayer culture protocol respectively for 30 days,then insulin expression was examined by RT-PCR,DTZ-staining and immunohistochemistry.The percentage of insulin-secreting cells was evaluated by flow cytometry.RESULTS: DTZ-staining positive cells and insulin immunohistochemical staining positive cells were observed in the differentiated cells for all the three groups.mRNAs of insulin and some other islet-related genes were detected,insulin expression was the strongest in EB formation-monolayer,and the weakest was in monolayer.The percentage of insulin-positive cells of the differentiated cells in the EB formation-monolayer group was higher than that in the EB formation group(P
