4.6 Article

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

期刊

CELLS
卷 10, 期 7, 页码 -

出版社

MDPI
DOI: 10.3390/cells10071671

关键词

myogenesis; skeletal muscle; differentiation; induced pluripotent stem cells (iPS); myogenic factors; CD56

资金

  1. National Science Centre in Poland [2013/09/B/NZ5/00769, 2018/29/B/NZ5/00915]

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Induced pluripotent stem cells are a useful tool for studying human embryo development processes such as myogenesis due to their ability to differentiate into three germ layers. Different protocols for obtaining myogenic cells have been described, with Protocol III found to be the most efficient. CD56 was also found to not be a specific marker for evaluating myogenic differentiation.
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-beta 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.

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