Skeletal and Cardiac Myogenesis Accompany Adipogenesis in P19 Embryonal Stem Cells
Skeletal and Cardiac Myogenesis Accompany Adipogenesis in P19 Embryonal Stem Cells
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DOI：
10.1089/scd.2008.0288
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发表时间：
2009-09-01
影响因子：
4
通讯作者：
Paquin, Joanne
中科院分区：
文献类型：
--
作者：
Bouchard, Frederic;Paquin, Joanne
P19胚胎癌细胞与正常胚胎干细胞相似。它们对维甲酸（RA）或催产素（OT）产生反应，产生心肌和骨骼肌细胞。RA处理后暴露于三碘甲状腺原氨酸（T3）和胰岛素诱导ES细胞分化为脂肪细胞和骨骼肌细胞。另一方面，据报道OT（10（-7）M）抑制3 T3前脂肪细胞成熟。目前的工作是进行，以确定是否P19细胞具有成脂潜力，可能会受到OT。用RA（10（-6）M）/T3+胰岛素（脂肪形成方案）或10（-7）M OT（心肌形成方案）处理细胞，并通过聚合酶链反应、免疫技术和细胞化学进行分析。油红-O染色和过氧化物酶体增殖物激活受体-γ（PPAR γ）和aP 2的表达表明在提交给脂肪形成方案的培养物中脂肪细胞的产生。还产生了收缩细胞。肌节辅肌动蛋白阳性和心肌肌钙蛋白抑制剂（cTnI）阴性的细胞表明生成骨骼肌细胞，cTnI阳性细胞表明生成心肌细胞。cTnI和骨骼标记物MyoD的水平在两种方案中几乎相似，而油红O染色与心肌原性方案无关。在成脂方案中加入10（-7）M OT不影响油红O染色和PPAR γ表达。有趣的是，Oct 3/4多能性标志物在脂肪形成方案中消失，但在心肌形成方案中仍然表达。因此，P19细胞具有不受10（-7）M OT影响的成脂潜能。RA/T3+胰岛素组合产生更大范围的中胚层细胞衍生物，是比OT更有效的形态发生治疗。P19细胞有助于研究发育过程中细胞命运决定的机制。
P19 embryonic carcinoma cells resemble normal embryonic stem (ES) cells. They generate cardiac and skeletal myocytes in response to retinoic acid (RA) or oxytocin (OT). RA treatment followed by exposure to triiodothyronine (T3) and insulin induces ES cells differentiation into adipocytes and skeletomyocytes. On the other hand, OT (10(-7) M) was reported to inhibit 3T3 preadipocyte maturation. The present work was undertaken to determine whether P19 cells have an adipogenic potential that could be affected by OT. Cells were treated with RA (10(-6) M)/T3+insulin (adipogenic protocol) or 10(-7) M OT (cardiomyogenic protocol), and analyzed by polymerase chain reaction, immunotechniques, and cytochemistry. Oil-Red-O staining and expression of peroxisome proliferator-activated receptor-gamma (PPAR gamma) and aP2 indicated the generation of adipocytes in cultures submitted to the adipogenic protocol. Contracting cells were also generated. Cells positive for sarcomeric actinin and negative for cardiac troponin inhibitor (cTpnI) indicated generation of skeletomyocytes, and cTpnI positive cells revealed generation of cardiomyocytes. Levels of cTpnI and of the skeletal marker MyoD were almost similar in both protocols, whereas no Oil-Red-O staining was associated with the cardiomyogenic protocol. Addition of 10(-7) M OT to the adipogenic protocol did not affect Oil-Red-O staining and PPAR gamma expression. Interestingly, Oct3/4 pluripotency marker disappeared in the adipogenic protocol but remained expressed in the cardiomyogenic one. P19 cells thus have an adipogenic potential non affected by 10(-7) M OT. RA/T3+insulin combination generates a larger spectrum of mesodermal cell derivatives and is a more potent morphogenic treatment than OT. P19 cells could help investigating mechanisms of cell fate decision during development.