Na?ve-like conversion of bovine induced pluripotent stem cells from Sertoli cells

Feeder cells are essential to derive pluripotent stem cells (PSCs). Mouse embryonic fibroblasts (MEF) are widely used as feeder to generate and culture embryonic stem cells (ESCs) and induced PSCs (iPSCs) in many species. However it may not be suitable for livestock ESCs/iPSCs due to interspecies difference. Previously we derived bovine iPSCs from bovine Sertoli cells using MEF feeder. Here we compared the effects of MEF feeder and bovine embryonic fibroblasts (BEF) feeder on the maintenance of bovine iPSC pluripotency and morphology as well their contributions to the naive-like conversion, based on a naive medium (NM). The results showed successful conversion of the primed bovine iPSCs to naive-like state within 3-4 days both on MEF feeder and BEF feeder in NM (termed as MNM and BNM respectively). These naive-like iPSCs showed normal karyotype. There were more iPSC colonies under BNM condition than MNM condition. Epigenetically, histone modification H3K4 was upregulated, while H3K27 was downregulated in the naive-like iPSCs. We further analyzed the naive markers and differentiation po-tential both in vitro and in vivo of these cells, which were all reserved throughout the maintenance. Together, bovine naive-like iPSCs can be generated both on MEF and BEF feeder in NM condition. The BNM condition is able to sustain the pluripotency and differentiation potential of the naive-like bovine iPSCs, and improve the conversion efficiency.(c) 2022 Elsevier Inc. All rights reserved.

Automated summary

This study compared the effects of mouse embryonic fibroblasts (MEF) and bovine embryonic fibroblasts (BEF) as feeder cells on the culture of bovine induced pluripotent stem cells (iPSCs). The results showed that both MEF and BEF feeder cells were able to successfully convert the bovine iPSCs to a naive-like state. The use of BEF feeder cells in the naive medium (BNM) condition resulted in better maintenance of pluripotency and differentiation potential, and improved conversion efficiency.