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iPS 细胞的成肌分化在方案的同时比较中显示出不同的效率。

Myogenic Differentiation of iPS Cells Shows Different Efficiency in Simultaneous Comparison of Protocols.

机构信息

Department of Transplantation, Medical College, Jagiellonian University, 31-008 Krakow, Poland.

Department of Transplantation, Institute of Pediatrics, Faculty of Medicine, Jagiellonian University Medical College, 30-663 Krakow, Poland.

出版信息

Cells. 2021 Jul 2;10(7):1671. doi: 10.3390/cells10071671.

DOI:10.3390/cells10071671
PMID:34359837
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8307201/
Abstract

Induced pluripotent stem (iPS) cells constitute a perfect tool to study human embryo development processes such as myogenesis, thanks to their ability to differentiate into three germ layers. Currently, many protocols to obtain myogenic cells have been described in the literature. They differ in many aspects, such as media components, including signaling modulators, feeder layer constituents, and duration of culture. In our study, we compared three different myogenic differentiation protocols to verify, side by side, their efficiency. Protocol I was based on embryonic bodies differentiation induction, ITS addition, and selection with adhesion to collagen I type. Protocol II was based on strong myogenic induction at the embryonic bodies step with BIO, forskolin, and bFGF, whereas cells in Protocol III were cultured in monolayers in three special media, leading to WNT activation and TGF-β and BMP signaling inhibition. Myogenic induction was confirmed by the hierarchical expression of myogenic regulatory factors MYF5, MYOD, MYF6 and MYOG, as well as the expression of myotubes markers MYH3 and MYH2, in each protocol. Our results revealed that Protocol III is the most efficient in obtaining myogenic cells. Furthermore, our results indicated that CD56 is not a specific marker for the evaluation of myogenic differentiation.

摘要

诱导多能干细胞（iPS）由于能够分化为三个胚层，因此构成了研究人类胚胎发育过程（如肌发生）的完美工具。目前，许多文献中已经描述了获得肌细胞的方法。它们在许多方面有所不同，例如培养基成分，包括信号调节剂、饲养层成分和培养时间。在我们的研究中，我们比较了三种不同的肌发生分化方案，以验证它们的效率。方案 I 基于胚胎体分化诱导、ITS 添加以及对 I 型胶原的黏附选择。方案 II 基于胚胎体阶段的强肌诱导，使用 BIO、forskolin 和 bFGF，而方案 III 中的细胞在三种特殊培养基中单层培养，导致 WNT 激活和 TGF-β 和 BMP 信号抑制。在每个方案中，通过肌生成调节因子 MYF5、MYOD、MYF6 和 MYOG 的分层表达以及肌管标志物 MYH3 和 MYH2 的表达来确认肌生成诱导。我们的结果表明，方案 III 是获得肌细胞最有效的方法。此外，我们的结果表明，CD56 不是评估肌生成分化的特异性标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/0f2d60f126a6/cells-10-01671-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/fd2f8987b4fd/cells-10-01671-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/3a986311955e/cells-10-01671-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/7e9763ae6522/cells-10-01671-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/a546a9bc04b9/cells-10-01671-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/0ff4b7b6df0b/cells-10-01671-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/0f2d60f126a6/cells-10-01671-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/fd2f8987b4fd/cells-10-01671-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/3a986311955e/cells-10-01671-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/7e9763ae6522/cells-10-01671-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/a546a9bc04b9/cells-10-01671-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/0ff4b7b6df0b/cells-10-01671-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7543/8307201/0f2d60f126a6/cells-10-01671-g006.jpg

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