Abstract
Large numbers of lipids exist in the porcine oocytes and early embryos and have the positive effects on their development, suggesting that the lipids may play an important role in pluripotency establishment and maintenance in pigs. However, the effects of lipids and their metabolites, such as fatty acids on reprogramming and the pluripotency gene expression of porcine-induced pluripotent stem cells (iPSCs), are unclear. Here, we generated the porcine iPSCs that resemble the mouse embryonic stem cells (ESCs) under lipid and fatty-acid-enriched cultural conditions (supplement of AlbuMAX). These porcine iPSCs show positive for the ESCs pluripotency markers and have the differentiation abilities to all three germ layers, and importantly, have the capability of aggregation into the inner cell mass (ICM) of porcine blastocysts. We further confirmed that lipid and fatty acid enriched condition can promote the cell proliferation and improve reprogramming efficiency by elevating cAMP levels. Interestingly, this lipids supplement promotes mesenchymal–epithelial transition (MET) through the cAMP/PKA/CREB signal pathway and upregulates the E-cadherin expression during porcine somatic cell reprogramming. The lipids supplement also makes a contribution to lipid droplets accumulation in the porcine iPSCs that resemble porcine preimplantation embryos. These findings may facilitate understanding of the lipid metabolism in porcine iPSCs and lay the foundation of bona fide porcine embryonic stem cell derivation.
Highlights
Induced pluripotent stem cells could be derived from somatic cell by the expression of transcription factors including OCT4, SOX2, KLF4 and c-MYC [1,2]
Our results of analysis of the transcriptome of the porcine early embryos at different development stages suggest that lipid and fatty acid metabolism may be a unique characteristic of porcine pre-implantation embryo development, which may play important roles in the cell fate regulation [4]
AlbuMAX is a kind of bovine serum albumin consisting of lipids and fatty acids, which are similar to lipid and fatty acid ingredients in porcine oocytes and early embryos
Summary
Induced pluripotent stem cells (iPSCs) could be derived from somatic cell by the expression of transcription factors including OCT4, SOX2, KLF4 and c-MYC [1,2]. Large amounts of lipids exist in porcine oocytes and early embryos, which play important roles in oocyte maturation, embryo development and stem cell proliferation [9]. Our results of analysis of the transcriptome of the porcine early embryos at different development stages suggest that lipid and fatty acid metabolism may be a unique characteristic of porcine pre-implantation embryo development, which may play important roles in the cell fate regulation [4]. Previous reports show that linoleic-acid-induced mouse embryonic stem cells (ESCs) proliferation through Ca2+/PKC, PI3K/Akt, and MAPK signaling pathways [10]. AlbuMAX improved the viability and hatching ability of porcine blastocysts produced in vitro [14], and regulated human embryonic stem cells (ESCs) self-renewal [15]. Sphere cells that have pluripotency could be induced from mouse fibroblasts in the culture conditions including AlbuMAX [16]
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