Timed inhibition of p38MAPK directs accelerated differentiation of human embryonic stem cells into cardiomyocytes.
Timed inhibition of p38MAPK directs accelerated differentiation of human embryonic stem cells into cardiomyocytes.
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DOI：
10.3109/14653249.2010.491821
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发表时间：
2010-10
期刊：
影响因子：
4.5
通讯作者：
Yeghiazarians Y
中科院分区：
文献类型：
--
作者：
Gaur M;Ritner C;Sievers R;Pedersen A;Prasad M;Bernstein HS;Yeghiazarians Y
人胚胎干细胞(HESC)来源的心肌细胞(HCM)的自发分化率低，限制了其治疗心力衰竭。由于已有研究表明p38丝裂原活化蛋白激酶(P38MAPK)可指导小鼠胚胎干细胞的神经发生，我们研究了p38MAPK抑制剂SB203580是否可能影响HCM的分化。在特定时间点用SB203580诱导分化人胚胎干细胞，并用流式细胞术、免疫细胞化学、实时定量聚合酶链式反应(RT-PCR)、畸胎瘤形成和透射电子显微镜观察心肌细胞的形成。我们观察到，在分化的第21天，抑制剂的加入使自发跳动的人胚样小体(Heb)增加了2.1倍，25%的处理细胞表达了心脏特异的α-肌球蛋白重链。这种作用依赖于引入抑制剂的分化阶段。免疫组织化学染色和畸胎瘤形成分析表明，该抑制物不影响人胚胎干细胞的多能性，但经处理的人胚胎干细胞诱导的人胚胎干细胞表现出肌节蛋白的表达增加，包括心肌肌钙蛋白T、肌球蛋白轻链和α-肌球蛋白重链。这与接受治疗的肥厚型心肌病早期显著增加的肌原纤维束数和新生Z小体的出现是一致的。经处理的Heb注射到小鼠心肌后，通过阵列比较基因组杂交和体内存活也证明了正常的核型。这些研究表明，抑制p38MAPK可以加速hESC向HCM定向分化，并且这种作用是发育阶段特有的。这种抑制剂的使用应该会提高我们产生hESC来源的HCM用于细胞治疗的能力。
Heart failure therapy with human embryonic stem cell (hESC)-derived cardiomyocytes (hCM) has been limited by the low rate of spontaneous hCM differentiation. As others have shown that p38 mitogen-activated protein kinase (p38MAPK) directs neurogenesis from mouse embryonic stem cells, we investigated whether the p38MAPK inhibitor, SB203580, might influence hCM differentiation. We treated differentiating hESC with SB203580 at specific time-points, and used flow cytometry, immunocytochemistry, quantitative real-time (RT)–polymerase chain reaction (PCR), teratoma formation and transmission electron microscopy to evaluate cardiomyocyte formation. We observed that the addition of inhibitor resulted in 2.1-fold enrichment of spontaneously beating human embryoid bodies (hEB) at 21 days of differentiation, and that 25% of treated cells expressed cardiac-specific α-myosin heavy chain. This effect was dependent on the stage of differentiation at which the inhibitor was introduced. Immunostaining and teratoma formation assays demonstrated that the inhibitor did not affect hESC pluripotency; however, treated hESC gave rise to hCM exhibiting increased expression of sarcomeric proteins, including cardiac troponin T, myosin light chain and α-myosin heavy chain. This was consistent with significantly increased numbers of myofibrillar bundles and the appearance of nascent Z-bodies at earlier time-points in treated hCM. Treated hEB also demonstrated a normal karyotype by array comparative genomic hybridization and viability in vivo following injection into mouse myocardium. These studies demonstrate that p38MAPK inhibition accelerates directed hCM differentiation from hESC, and that this effect is developmental stage-specific. The use of this inhibitor should improve our ability to generate hESC-derived hCM for cell-based therapy.
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影响因子：
3.9
作者：
Bin, Zeng;Sheng, Lin-Guo;Hui, Shen
通讯作者：
Hui, Shen
DOI：
10.1152/ajpheart.00020.2003
发表时间：
2003-12-01
影响因子：
4.8
作者：
Snir, M;Kehat, I;Gepstein, L
通讯作者：
Gepstein, L
影响因子：
20.1
作者：
Kehat, I;Gepstein, A;Gepstein, L
通讯作者：
Gepstein, L
影响因子：
6
作者：
Laflamme, MA;Gold, J;Murry, CE
通讯作者：
Murry, CE
影响因子：
24
作者：
Caspi, Oren;Huber, Irit;Gepstein, Lior
通讯作者：
Gepstein, Lior