Comparison of the effects on differentiation of mouse embryonic stem cells into insulin-secreting cells among three cell culture protocols / 中国病理生理杂志

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