Pyruvate and recombinant human basic fibroblast growth factor rescue the pluripotency and metabolism of bovine embryonic stem cells on low-density feeder layers

Feeder cells still play an important role in the in vitro culture of bovine embryonic stem cells(bESCs). A suitable microenvironment or niche is essential for the self-renewal and pluripotency of stem cells cultured in vitro. Feeder cells participate in the construction of stem cell niches by secreting growth factors and extracellular matrix proteins. In this study, we used metabolomics and transcriptomics to investigate the effects of feeder layer deficiency on bESCs. bESCs co-cultured with low-density feeder cells experienced a decrease in pluripotent gene expression, cell differentiation, and central carbon metabolic activity. When cell-permeable pyruvate (Pyr) and recombinant human basic fibroblast growth factor (rhbFGF) were added to the culture system, the pluripotency of bESCs on low-density feeder layers was rescued, and acetyl-coenzyme A(AcCoA) synthesis and fatty acid de novo synthesis increased. rhbFGF enhances the effects of Pyr and activates the overall metabolic level of bESCs grown on low-density feeder layers. Overall design: Pluripotent stem cells mRNA profiles in MC-F7,1/7MC-F7,1/7MC+Pyr-F7,1/7MC+Pyr+FGF-F7. Note,1M equal to MC, 1M-F7 equal to MC-F7, 1/7M-F7 equal to 1/7MC-F7.