Hair-follicle-associated pluripotent (HAP) stem cells differentiate into mature beating cardiomyocyte sheets on flexible substrates in vitro
Hair-follicle-associated pluripotent (HAP) stem cells differentiate into mature beating cardiomyocyte sheets on flexible substrates in vitro
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DOI：
10.1007/s00795-022-00322-z
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发表时间：
2022-05-10
影响因子：
1.8
通讯作者：
Amoh, Yasuyuki
中科院分区：
文献类型：
--
作者：
Takaoka, Nanako;Yamane, Michiko;Amoh, Yasuyuki
心肌细胞已经从各种干细胞如人胚胎干细胞（ESC）和诱导多能干细胞（iPSC）分化而来，但是难以产生成熟的心肌细胞。我们发现，大鼠毛囊相关多能（HAP）干细胞具有多能性，并产生成熟的搏动心肌细胞片从大鼠HAP干细胞分化。将大鼠触须毛囊上部培养在10%FBS DMEM中，用外胚层、中胚层、内胚层系统的抗体染色，显示多种细胞类型的分化。此外，在三种不同的条件下培养HAP干细胞，以确定最合适的培养条件，使跳动的心肌细胞片。肌节结构染色显示搏动的心肌细胞片层成熟。单独的异丙肾上腺素和异丙肾上腺素，激活素A，骨形态发生蛋白4（BMP 4）和碱性成纤维细胞生长因子（bFGF）的组合有效地诱导跳动的长纤维心肌细胞，形成跳动片，只有在所有四种药物的存在下。柔性基质对于HAP干细胞的成熟搏动心肌细胞片的分化是必不可少的。由HAP干细胞分化的心肌细胞的特征表明它们具有心脏再生的临床潜力。
Cardiomyocytes have been differentiated from various stem cells such as human embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC), but it is difficult to produce mature cardiomyocytes. We showed rat hair-follicle-associated pluripotent (HAP) stem cells have pluripotency and produced mature beating cardiomyocyte sheets differentiated from rat HAP stem cells. The upper parts of rat vibrissa hair follicles were cultured in 10% FBS DMEM and stained with antibodies of the ectoderm, mesoderm, endoderm system to show the differentiation of multiple cell types. Moreover, HAP stem cells were cultured under three different conditions to decide the most suitable culture conditions for making beating cardiomyocyte sheets. The beating cardiomyocyte sheets were shown to be mature by staining sarcomere structures. Isoproterenol alone and the combination of isoproterenol, activin A, bone morphogenetic protein 4 (BMP4) and basic fibroblast growth factor (bFGF) effectively induced beating long-fiber cardiomyocytes, which formed beating sheets, only in the presence of all four agents. Flexible substrates were essential for the differentiation of sheets of mature beating cardiomyocytes for HAP stem cells. The features of the cardiomyocytes differentiated from HAP stem cells demonstrate they have clinical potential for heart regeneration.