Serum and non-serum method-induced differentiation of mouse embryonic stem cells into definitive endoderm / 第二军医大学学报

ABSTRACT

Objective: To compare the efficiencies of two methods (serum and non-serum) in inducing mouse embryonic stem cell differentiation into definitive endoderm cells. Methods: The serum and non-serum methods were used to induce differentiation of mouse embryonic stem cells into definitive endoderm cells. Fluorescence activated cell sorter (FACS) was used to analyze the inducing time and efficiency of definitive endoderm using their surface protein marks (Cxcr4, c-Kit and E-cadherin). Meanwhile, RT-PCR was used to analyze the gene profile of definitive endoderm induced by the two methods. Realtime PCR was used to analyze the gene expression in definitive endoderm during the induction course in the non-serum group. The Cxcr4 and c-Kit double positive definitive endoderm cells were sorted by flow cytometry and gene profile was characterized by RT-PCR. Results: Definitive endoderm cells were induced from mouse embryonic stem cells by both serum and non-serum methods. However, the efficiency of non-serum group (74. 19%) was higher than that in the serum group, and the induction outcome reached a climax at the 4th day of induction. Conclusion: We have established a highly efficient method to induce differentiation of mouse embryonic stem cells into definitive endoderm, which lays a foundation for further differentiation into liver and pancreatic cells.