title: Phosphatidic Acid Improves Reprogramming to Pluripotency by Reducing Apoptosis.
abstract: Generation of induced pluripotent stem cells (iPSCs) requires a considerable amount of lipids, such as phosphatidic acid (PA), to meet the needs of subsequent rapid cell division and proliferation. However, it is unclear whether PA, a biosynthetic precursor of lipids, affects reprogramming. By using lentiviral expression of the Yamanaka factors in mouse embryonic fibroblasts for reprogramming, we identified that PA is beneficial for the generation of iPS colonies. Inhibiting the generation of cellular PA dramatically decreased the number of iPSCs. Consistently, 400 μM PA improved iPSC generation by more than 4- to 5-fold. iPSCs generated in the presence of PA (PA-iPS) expressed pluripotent markers such as Oct4 and Nanog, differentiated into cells of the three germ layers in vitro, and contributed to chimeric mice when injected into blastocysts. The improved efficiency was primarily due to reduction of apoptosis as sufficient PA increased the accumulation of cardiolipin in the inner membrane of the mitochondria, which reduced the release of cytochrome c and, in turn, suppressed apoptosis by inhibiting caspase-7. The relatively higher amount of Bcl-2 in PA treatment also inhibited apoptosis. In addition, an accompanied sequential change from epithelial-to-mesenchymal transition (EMT) at the initial phase of reprogramming to mesenchymal-to-epithelial transition (MET) was also detected. Our microarray data, which also supported our results, indicated the presence of significant membrane enrichment genes, thus suggesting that PA may function through membrane-anchored proteins. We thus identified a novel type of culture supplement that improves the efficiency of reprogramming and could be valuable for the generation of high-quality iPS cells.
authors.affiliation: 1 The State Key Laboratory of Reproductive Biology, Institute of Zoology , Chinese Academy of Sciences, Beijing, China .|2 University of Chinese Academy of Sciences , Beijing, China .
authors.forename: Yuan
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: 1 The State Key Laboratory of Reproductive Biology, Institute of Zoology , Chinese Academy of Sciences, Beijing, China .|3 School of Life Sciences, Anhui University , Hefei, China .
authors.forename: Mingxia
authors.initials: M
authors.lastname: Du
authors.affiliation: 1 The State Key Laboratory of Reproductive Biology, Institute of Zoology , Chinese Academy of Sciences, Beijing, China .|2 University of Chinese Academy of Sciences , Beijing, China .
authors.forename: Menghua
authors.initials: M
authors.lastname: Wu
authors.affiliation: 1 The State Key Laboratory of Reproductive Biology, Institute of Zoology , Chinese Academy of Sciences, Beijing, China .|2 University of Chinese Academy of Sciences , Beijing, China .
authors.forename: Yanbing
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: 1 The State Key Laboratory of Reproductive Biology, Institute of Zoology , Chinese Academy of Sciences, Beijing, China .|2 University of Chinese Academy of Sciences , Beijing, China .
authors.forename: Xing
authors.initials: X
authors.lastname: Zhao
authors.affiliation: 1 The State Key Laboratory of Reproductive Biology, Institute of Zoology , Chinese Academy of Sciences, Beijing, China .|2 University of Chinese Academy of Sciences , Beijing, China .
authors.forename: Xu
authors.initials: X
authors.lastname: Cao
authors.affiliation: 1 The State Key Laboratory of Reproductive Biology, Institute of Zoology , Chinese Academy of Sciences, Beijing, China .|2 University of Chinese Academy of Sciences , Beijing, China .
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.affiliation: 1 The State Key Laboratory of Reproductive Biology, Institute of Zoology , Chinese Academy of Sciences, Beijing, China .|2 University of Chinese Academy of Sciences , Beijing, China .
authors.forename: Peipei
authors.initials: P
authors.lastname: Long
authors.affiliation: 1 The State Key Laboratory of Reproductive Biology, Institute of Zoology , Chinese Academy of Sciences, Beijing, China .|2 University of Chinese Academy of Sciences , Beijing, China .
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: 1 The State Key Laboratory of Reproductive Biology, Institute of Zoology , Chinese Academy of Sciences, Beijing, China .|2 University of Chinese Academy of Sciences , Beijing, China .
authors.forename: Baoyang
authors.initials: B
authors.lastname: Hu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000187:drug effects*; D000067470:Cellular Reprogramming Techniques; D015536:Down-Regulation / Q000187:drug effects; D004622:Embryo, Mammalian; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D010712:Phosphatidic Acids / Q000494:pharmacology*
chemical_list: D010712:Phosphatidic Acids