This article aims to investigate the roles of differential metabolites pantothenic acid(PA)and fatty acid oxidation(FAO)in the differentiation of chicken embryonic stem cells(ESCs)into primor-dial germ cells(PGCs),and to provide a new theoretical basis for analyzing the mechanism of PGCs for-mation from a metabolic perspective.Using BMP4 induction system,five groups were established based on the optimal concentrations of pantothenic acid(PA)and its inhibitor(PZ)and fatty acid oxidation activa-tor(BMS)/inhibitor(Per)in previous studies of the research group:Control,PA+BMS,PA+Per,PZ+BMS,and PZ+Per.Cell morphology was observed,PGCs-specific marker genes were detected by qRT-PCR,and indirect immunofluorescence and flow cytometry were employed to explore the effects of panto-thenate metabolism and its downstream fatty acid oxidation during ESCs to PGCs differentiation.Cell mor-phology observations at day 6 of induction showed a significant increase in embryoid bodies in the PZ+BMS group(P＜0.01)and a significant decrease in the PA+Per group(P＜0.05).qRT-PCR results indi-cated a significant increase in CVH and C-KIT expression in the PZ+BMS-induced group(P＜0.01),while their expression was significantly reduced in the PA+Per group(P＜0.01).Indirect immunofluores-cence and flow cytometric sorting results demonstrated a significantly higher proportion of DDX4-positive cells in the PZ+BMS group compared to the control(P＜0.05),whereas the PA+Per group showed a significantly lower proportion(P＜0.01).These findings suggest that inhibiting pantothenic acid metabo-lism and activating fatty acid metabolism can promote PGCs formation.This study provides new insights into the optimization of in vitro induction culture system of PGCs using metabolites.
维普
万方数据