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

Overview

Journal Cells

Publisher MDPI

Specialties Biochemistry
Biophysics
Cell Biology
Molecular Biology

Date 2021 Aug 7

PMID 34359837

Citations 3

Authors

Aleksandra Ulman

Marta Kot

Klaudia Skrzypek

Barbara Szewczyk

Marcin Majka

Affiliations

Soon will be listed here.

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.

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References

Tortorella L, Milasincic D, Pilch P . Critical proliferation-independent window for basic fibroblast growth factor repression of myogenesis via the p42/p44 MAPK signaling pathway. J Biol Chem. 2001; 276(17):13709-17. DOI: 10.1074/jbc.M100091200. View

Sanchez-Gurmaches J, Guertin D . Adipocytes arise from multiple lineages that are heterogeneously and dynamically distributed. Nat Commun. 2014; 5:4099. PMC: 4066194. DOI: 10.1038/ncomms5099. View

Kim J, Magli A, Chan S, Oliveira V, Wu J, Darabi R . Expansion and Purification Are Critical for the Therapeutic Application of Pluripotent Stem Cell-Derived Myogenic Progenitors. Stem Cell Reports. 2017; 9(1):12-22. PMC: 5511038. DOI: 10.1016/j.stemcr.2017.04.022. View

Xu C, Tabebordbar M, Iovino S, Ciarlo C, Liu J, Castiglioni A . A zebrafish embryo culture system defines factors that promote vertebrate myogenesis across species. Cell. 2013; 155(4):909-921. PMC: 3902670. DOI: 10.1016/j.cell.2013.10.023. View

Takada S, Stark K, Shea M, Vassileva G, McMahon J, McMahon A . Wnt-3a regulates somite and tailbud formation in the mouse embryo. Genes Dev. 1994; 8(2):174-89. DOI: 10.1101/gad.8.2.174. View

Relaix F, Rocancourt D, Mansouri A, Buckingham M . A Pax3/Pax7-dependent population of skeletal muscle progenitor cells. Nature. 2005; 435(7044):948-53. DOI: 10.1038/nature03594. View

Chlebanowska P, Tejchman A, Sulkowski M, Skrzypek K, Majka M . Use of 3D Organoids as a Model to Study Idiopathic Form of Parkinson's Disease. Int J Mol Sci. 2020; 21(3). PMC: 7037292. DOI: 10.3390/ijms21030694. View

Borchin B, Chen J, Barberi T . Derivation and FACS-mediated purification of PAX3+/PAX7+ skeletal muscle precursors from human pluripotent stem cells. Stem Cell Reports. 2013; 1(6):620-31. PMC: 3871395. DOI: 10.1016/j.stemcr.2013.10.007. View

Chal J, Oginuma M, Al Tanoury Z, Gobert B, Sumara O, Hick A . Differentiation of pluripotent stem cells to muscle fiber to model Duchenne muscular dystrophy. Nat Biotechnol. 2015; 33(9):962-9. DOI: 10.1038/nbt.3297. View

10.

Darabi R, Perlingeiro R . Lineage-specific reprogramming as a strategy for cell therapy. Cell Cycle. 2008; 7(12):1732-7. DOI: 10.4161/cc.7.12.6159. View

11.

Ott M, Bober E, Lyons G, Arnold H, Buckingham M . Early expression of the myogenic regulatory gene, myf-5, in precursor cells of skeletal muscle in the mouse embryo. Development. 1991; 111(4):1097-107. DOI: 10.1242/dev.111.4.1097. View

12.

Mendez M, Kojima S, Goldman R . Vimentin induces changes in cell shape, motility, and adhesion during the epithelial to mesenchymal transition. FASEB J. 2010; 24(6):1838-51. PMC: 2874471. DOI: 10.1096/fj.09-151639. View

13.

Schiaffino S, Rossi A, Smerdu V, Leinwand L, Reggiani C . Developmental myosins: expression patterns and functional significance. Skelet Muscle. 2015; 5:22. PMC: 4502549. DOI: 10.1186/s13395-015-0046-6. View

14.

Jiwlawat N, Lynch E, Jeffrey J, Van Dyke J, Suzuki M . Current Progress and Challenges for Skeletal Muscle Differentiation from Human Pluripotent Stem Cells Using Transgene-Free Approaches. Stem Cells Int. 2018; 2018:6241681. PMC: 5924987. DOI: 10.1155/2018/6241681. View

15.

Chal J, Pourquie O . Making muscle: skeletal myogenesis and . Development. 2017; 144(12):2104-2122. DOI: 10.1242/dev.151035. View

16.

Skrzypek K, Kusienicka A, Trzyna E, Szewczyk B, Ulman A, Konieczny P . SNAIL is a key regulator of alveolar rhabdomyosarcoma tumor growth and differentiation through repression of MYF5 and MYOD function. Cell Death Dis. 2018; 9(6):643. PMC: 5974324. DOI: 10.1038/s41419-018-0693-8. View

17.

Musial-Wysocka A, Kot M, Sulkowski M, Badyra B, Majka M . Molecular and Functional Verification of Wharton's Jelly Mesenchymal Stem Cells (WJ-MSCs) Pluripotency. Int J Mol Sci. 2019; 20(8). PMC: 6515095. DOI: 10.3390/ijms20081807. View

18.

Shelton M, Kocharyan A, Liu J, Skerjanc I, Stanford W . Robust generation and expansion of skeletal muscle progenitors and myocytes from human pluripotent stem cells. Methods. 2015; 101:73-84. DOI: 10.1016/j.ymeth.2015.09.019. View

19.

Ulman A, Skrzypek K, Konieczny P, Mussolino C, Cathomen T, Majka M . Genome Editing of the Gene in Rhabdomyosarcoma: A Novel Model for Studies of Its Role. Cells. 2020; 9(5). PMC: 7290443. DOI: 10.3390/cells9051095. View

20.

Yamaguchi T, Harpal K, Henkemeyer M, Rossant J . fgfr-1 is required for embryonic growth and mesodermal patterning during mouse gastrulation. Genes Dev. 1994; 8(24):3032-44. DOI: 10.1101/gad.8.24.3032. View