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Cellular reprogramming2018; 20(5); 275-281; doi: 10.1089/cell.2018.0023

Generation of Functional Myocytes from Equine Induced Pluripotent Stem Cells.

Abstract: Induced pluripotent stem cells (iPSCs) have revolutionized human biomedicine through their use in disease modeling and therapy. In comparison, little progress has been made toward the application of iPSCs in veterinary species. In that regard, skeletal myocytes from iPSCs would have great potential for understanding muscle function and disease in the equine athlete. In this study, we generated skeletal myotubes by transducing equine iPSC-derived mesenchymal derivatives with an inducible lentiviral vector coding for the human sequence of the myogenic factor, MyoD. Myosin heavy chain-positive myotubes generated from two different iPSC lines were compared to myotubes from adult equine skeletal muscle progenitor cells (MPCs). iPSC myotubes had a smaller mean area than MPC myotubes (≤2-fold). In addition, quantitative polymerase chain reaction analyses showed that iPSC myotubes expressed MYH2 and MYH3 isoforms (at similar or lower levels than MPC myotubes), but they did not express the mature muscle isoform, MYH1. Compared to MPC myotubes, iPSC myotubes expressed reduced levels of the myogenic factors, MYOD1 and MYF6, but did not express MYF5. Finally, iPSC myotubes responded to KCl-induced membrane depolarization by releasing calcium and did so in a manner similar to MPC myotubes. In conclusion, this is the first study to report the generation of functional myocytes from equine iPSCs.
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Publication Date: 2018-09-12 PubMed ID: 30207795PubMed Central: PMC6166488DOI: 10.1089/cell.2018.0023Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study focuses on the creation of functional muscle cells (myocytes) from horse-derived stem cells (also known as Equine Induced Pluripotent Stem Cells) and is regarded as a significant stride towards understanding muscle function and potential diseases in horses.

Introduction

  • The current research revolves around Induced Pluripotent Stem Cells (iPSCs) that have brought about a revolution in human biomedicine due to their application in disease modeling and therapy.
  • However, their application in veterinary species hasn’t seen much progress, hence this research attempts to use these iPSCs to generate skeletal muscle cells, which could be a significant leap in understanding muscle function and diseases in horses.

Method

  • The researchers induced skeletal muscle tube formation by transducing horse-derived iPSC mesenchymal derivatives using a lentiviral vector that codes for the human sequence of the muscle-forming factor, MyoD.
  • They then compared these iPSC-created myotubes with myotubes developed from adult horse skeletal muscle progenitor cells (MPCs).

Findings

  • The myotubes generated from iPSCs had a smaller mean area compared to those created from MPCs (≤2-fold).
  • Quantitative polymerase chain reaction analyses revealed that the iPSC myotubes expressed MYH2 and MYH3 isoforms at levels comparable or lower than the MPC myotubes, but they didn’t express the mature muscle isoform, MYH1.
  • iPSC myotubes had lower levels of the muscle-forming factors, MYOD1 and MYF6, but did not express MYF5, in comparison to MPC myotubes.
  • Interestingly, the iPSC myotubes responded to KCl-induced membrane depolarization by releasing calcium in a similar manner to the MPC myotubes.

Conclusion

  • This research claims to be the first to report the creation of functional myocytes from horse-derived stem cells (iPSCs).
  • Such study further exhibits the potential uses of iPSCs not just in human biology and medicine but also in the veterinary field.

Cite This Article

APA
Amilon KR, Cortes-Araya Y, Moore B, Lee S, Lillico S, Breton A, Esteves CL, Donadeu FX. (2018). Generation of Functional Myocytes from Equine Induced Pluripotent Stem Cells. Cell Reprogram, 20(5), 275-281. https://doi.org/10.1089/cell.2018.0023

Publication

Cellular reprogramming
ISSN: 2152-4998
NlmUniqueID: 101528176
Country: United States
Language: English
Volume: 20
Issue: 5
Pages: 275-281

Researcher Affiliations

Amilon, Karin R
  • 1 The Roslin Institute and R(D)SVS, University of Edinburgh , Edinburgh, United Kingdom .
Cortes-Araya, Yennifer
  • 1 The Roslin Institute and R(D)SVS, University of Edinburgh , Edinburgh, United Kingdom .
Moore, Benjamin
  • 1 The Roslin Institute and R(D)SVS, University of Edinburgh , Edinburgh, United Kingdom .
Lee, Seungmee
  • 1 The Roslin Institute and R(D)SVS, University of Edinburgh , Edinburgh, United Kingdom .
Lillico, Simon
  • 1 The Roslin Institute and R(D)SVS, University of Edinburgh , Edinburgh, United Kingdom .
Breton, Amandine
  • 1 The Roslin Institute and R(D)SVS, University of Edinburgh , Edinburgh, United Kingdom .
Esteves, Cristina L
  • 1 The Roslin Institute and R(D)SVS, University of Edinburgh , Edinburgh, United Kingdom .
Donadeu, F Xavier
  • 1 The Roslin Institute and R(D)SVS, University of Edinburgh , Edinburgh, United Kingdom .
  • 2 The Euan Macdonald Centre for Motor Neurone Disease Research, University of Edinburgh , Edinburgh, United Kingdom .

MeSH Terms

  • Animals
  • Cell Differentiation
  • Cells, Cultured
  • Horses
  • Induced Pluripotent Stem Cells / cytology
  • Muscle Fibers, Skeletal / cytology
  • Muscle, Skeletal / cytology

Grant Funding

  • BBS/E/D/20221658 / Biotechnology and Biological Sciences Research Council
  • BBS/E/D/10002071 / Biotechnology and Biological Sciences Research Council

Conflict of Interest Statement

The authors declare they have no financial conflicts of interest.

References

This article includes 23 references
  1. Albini S, Coutinho P, Malecova B, Giordani L, Savchenko A, Forcales SV, Puri PL. Epigenetic reprogramming of human embryonic stem cells into skeletal muscle cells and generation of contractile myospheres.. Cell Rep 2013 Mar 28;3(3):661-70.
    pmc: PMC3625045pubmed: 23478022doi: 10.1016/j.celrep.2013.02.012google scholar: lookup
  2. Baquero-Perez B, Kuchipudi SV, Nelli RK, Chang KC. A simplified but robust method for the isolation of avian and mammalian muscle satellite cells.. BMC Cell Biol 2012 Jun 21;13:16.
    pmc: PMC3432597pubmed: 22720831doi: 10.1186/1471-2121-13-16google scholar: lookup
  3. Bentzinger CF, Wang YX, Rudnicki MA. Building muscle: molecular regulation of myogenesis.. Cold Spring Harb Perspect Biol 2012 Feb 1;4(2).
    pmc: PMC3281568pubmed: 22300977doi: 10.1101/cshperspect.a008342google scholar: lookup
  4. Breton A, Sharma R, Diaz AC, Parham AG, Graham A, Neil C, Whitelaw CB, Milne E, Donadeu FX. Derivation and characterization of induced pluripotent stem cells from equine fibroblasts.. Stem Cells Dev 2013 Feb 15;22(4):611-21.
    pmc: PMC3564467pubmed: 22897112doi: 10.1089/scd.2012.0052google scholar: lookup
  5. Chal J, Al Tanoury Z, Hestin M, Gobert B, Aivio S, Hick A, Cherrier T, Nesmith AP, Parker KK, Pourquié O. Generation of human muscle fibers and satellite-like cells from human pluripotent stem cells in vitro.. Nat Protoc 2016 Oct;11(10):1833-50.
    pubmed: 27583644doi: 10.1038/nprot.2016.110google scholar: lookup
  6. Chal J, Pourquié O. Making muscle: skeletal myogenesis in vivo and in vitro.. Development 2017 Jun 15;144(12):2104-2122.
    pubmed: 28634270doi: 10.1242/dev.151035google scholar: lookup
  7. Chen WC, Baily JE, Corselli M, Díaz ME, Sun B, Xiang G, Gray GA, Huard J, Péault B. Human myocardial pericytes: multipotent mesodermal precursors exhibiting cardiac specificity.. Stem Cells 2015 Feb;33(2):557-73.
    pmc: PMC4762368pubmed: 25336400doi: 10.1002/stem.1868google scholar: lookup
  8. Choi IY, Lim H, Estrellas K, Mula J, Cohen TV, Zhang Y, Donnelly CJ, Richard JP, Kim YJ, Kim H, Kazuki Y, Oshimura M, Li HL, Hotta A, Rothstein J, Maragakis N, Wagner KR, Lee G. Concordant but Varied Phenotypes among Duchenne Muscular Dystrophy Patient-Specific Myoblasts Derived using a Human iPSC-Based Model.. Cell Rep 2016 Jun 7;15(10):2301-2312.
    pubmed: 27239027doi: 10.1016/j.celrep.2016.05.016google scholar: lookup
  9. Darabi R, Arpke RW, Irion S, Dimos JT, Grskovic M, Kyba M, Perlingeiro RC. Human ES- and iPS-derived myogenic progenitors restore DYSTROPHIN and improve contractility upon transplantation in dystrophic mice.. Cell Stem Cell 2012 May 4;10(5):610-9.
    pmc: PMC3348507pubmed: 22560081doi: 10.1016/j.stem.2012.02.015google scholar: lookup
  10. Donadeu FX. Equine induced pluripotent stem cells or how to turn skin cells into neurons: horse tissues a la carte?. Equine Vet J 2014 Sep;46(5):534-7.
    pubmed: 25099189doi: 10.1111/evj.12300google scholar: lookup
  11. Fernandez-Fuente M, Ames EG, Wagner ML, Zhou H, Strom M, Zammit PS, Mickelson JR, Muntoni F, Brown SC, Piercy RJ. Assessment of the transformation of equine skin-derived fibroblasts to multinucleated skeletal myotubes following lentiviral-induced expression of equine myogenic differentiation 1.. Am J Vet Res 2008 Dec;69(12):1637-45.
    pubmed: 19046012doi: 10.2460/ajvr.69.12.1637google scholar: lookup
  12. Goudenege S, Lebel C, Huot NB, Dufour C, Fujii I, Gekas J, Rousseau J, Tremblay JP. Myoblasts derived from normal hESCs and dystrophic hiPSCs efficiently fuse with existing muscle fibers following transplantation.. Mol Ther 2012 Nov;20(11):2153-67.
    pmc: PMC3498803pubmed: 22990676doi: 10.1038/mt.2012.188google scholar: lookup
  13. Hockemeyer D, Jaenisch R. Induced Pluripotent Stem Cells Meet Genome Editing.. Cell Stem Cell 2016 May 5;18(5):573-86.
    pmc: PMC4871596pubmed: 27152442doi: 10.1016/j.stem.2016.04.013google scholar: lookup
  14. Iovino S, Burkart AM, Warren L, Patti ME, Kahn CR. Myotubes derived from human-induced pluripotent stem cells mirror in vivo insulin resistance.. Proc Natl Acad Sci U S A 2016 Feb 16;113(7):1889-94.
    pmc: PMC4763759pubmed: 26831110doi: 10.1073/pnas.1525665113google scholar: lookup
  15. Kabadi AM, Thakore PI, Vockley CM, Ousterout DG, Gibson TM, Guilak F, Reddy TE, Gersbach CA. Enhanced MyoD-induced transdifferentiation to a myogenic lineage by fusion to a potent transactivation domain.. ACS Synth Biol 2015 Jun 19;4(6):689-99.
    pmc: PMC4475448pubmed: 25494287doi: 10.1021/sb500322ugoogle scholar: lookup
  16. Lee EM, Kim AY, Lee EJ, Park JK, Park SI, Cho SG, Kim HK, Kim SY, Jeong KS. Generation of Equine-Induced Pluripotent Stem Cells and Analysis of Their Therapeutic Potential for Muscle Injuries.. Cell Transplant 2016 Nov;25(11):2003-2016.
    pubmed: 27226077doi: 10.3727/096368916X691691google scholar: lookup
  17. Mickelson JR, Valberg SJ. The genetics of skeletal muscle disorders in horses.. Annu Rev Anim Biosci 2015;3:197-217.
    pubmed: 25387114doi: 10.1146/annurev-animal-022114-110653google scholar: lookup
  18. Naylor RJ, Piercy RJ. Development of a clonal equine myoblast cell line capable of terminal differentiation into mature myotubes in vitro.. Am J Vet Res 2015 Jul;76(7):608-14.
    pubmed: 26111090doi: 10.2460/ajvr.76.7.608google scholar: lookup
  19. Quattrocelli M, Giacomazzi G, Broeckx SY, Ceelen L, Bolca S, Spaas JH, Sampaolesi M. Equine-Induced Pluripotent Stem Cells Retain Lineage Commitment Toward Myogenic and Chondrogenic Fates.. Stem Cell Reports 2016 Jan 12;6(1):55-63.
    pmc: PMC4719186pubmed: 26771353doi: 10.1016/j.stemcr.2015.12.005google scholar: lookup
  20. Schiaffino S, Rossi AC, Smerdu V, Leinwand LA, Reggiani C. Developmental myosins: expression patterns and functional significance.. Skelet Muscle 2015;5:22.
    pmc: PMC4502549pubmed: 26180627doi: 10.1186/s13395-015-0046-6google scholar: lookup
  21. Sharma R, Livesey MR, Wyllie DJ, Proudfoot C, Whitelaw CB, Hay DC, Donadeu FX. Generation of functional neurons from feeder-free, keratinocyte-derived equine induced pluripotent stem cells.. Stem Cells Dev 2014 Jul 1;23(13):1524-34.
    pubmed: 24548115doi: 10.1089/scd.2013.0565google scholar: lookup
  22. Skoglund G, Lainé J, Darabi R, Fournier E, Perlingeiro R, Tabti N. Physiological and ultrastructural features of human induced pluripotent and embryonic stem cell-derived skeletal myocytes in vitro.. Proc Natl Acad Sci U S A 2014 Jun 3;111(22):8275-80.
    pmc: PMC4050534pubmed: 24843168doi: 10.1073/pnas.1322258111google scholar: lookup
  23. Xu C, Tabebordbar M, Iovino S, Ciarlo C, Liu J, Castiglioni A, Price E, Liu M, Barton ER, Kahn CR, Wagers AJ, Zon LI. A zebrafish embryo culture system defines factors that promote vertebrate myogenesis across species.. Cell 2013 Nov 7;155(4):909-921.
    pmc: PMC3902670pubmed: 24209627doi: 10.1016/j.cell.2013.10.023google scholar: lookup

Citations

This article has been cited 10 times.
  1. Barrachina L, Arshaghi TE, O'Brien A, Ivanovska A, Barry F. Induced pluripotent stem cells in companion animals: how can we move the field forward?. Front Vet Sci 2023;10:1176772.
    doi: 10.3389/fvets.2023.1176772pubmed: 37180067google scholar: lookup
  2. Rooney MF, Neto NGB, Monaghan MG, Hill EW, Porter RK. Conditionally immortalised equine skeletal muscle cell lines for in vitro analysis. Biochem Biophys Rep 2023 Mar;33:101391.
    doi: 10.1016/j.bbrep.2022.101391pubmed: 36504704google scholar: lookup
  3. Knežić T, Janjušević L, Djisalov M, Yodmuang S, Gadjanski I. Using Vertebrate Stem and Progenitor Cells for Cellular Agriculture, State-of-the-Art, Challenges, and Future Perspectives. Biomolecules 2022 May 13;12(5).
    doi: 10.3390/biom12050699pubmed: 35625626google scholar: lookup
  4. Scarfone RA, Pena SM, Russell KA, Betts DH, Koch TG. The use of induced pluripotent stem cells in domestic animals: a narrative review. BMC Vet Res 2020 Dec 8;16(1):477.
    doi: 10.1186/s12917-020-02696-7pubmed: 33292200google scholar: lookup
  5. Gheibi S, Singh T, da Cunha JPMCM, Fex M, Mulder H. Insulin/Glucose-Responsive Cells Derived from Induced Pluripotent Stem Cells: Disease Modeling and Treatment of Diabetes. Cells 2020 Nov 12;9(11).
    doi: 10.3390/cells9112465pubmed: 33198288google scholar: lookup
  6. Su Y, Zhu J, Salman S, Tang Y. Induced pluripotent stem cells from farm animals. J Anim Sci 2020 Nov 1;98(11).
    doi: 10.1093/jas/skaa343pubmed: 33098420google scholar: lookup
  7. Jin Y, Shen Y, Su X, Weintraub NL, Tang Y. Effective restoration of dystrophin expression in iPSC (Mdx)-derived muscle progenitor cells using the CRISPR/Cas9 system and homology-directed repair technology. Comput Struct Biotechnol J 2020;18:765-773.
    doi: 10.1016/j.csbj.2020.03.012pubmed: 32280431google scholar: lookup
  8. Pessôa LVF, Bressan FF, Freude KK. Induced pluripotent stem cells throughout the animal kingdom: Availability and applications. World J Stem Cells 2019 Aug 26;11(8):491-505.
    doi: 10.4252/wjsc.v11.i8.491pubmed: 31523369google scholar: lookup
  9. Chung MJ, Park S, Son JY, Lee JY, Yun HH, Lee EJ, Lee EM, Cho GJ, Lee S, Park HS, Jeong KS. Differentiation of equine induced pluripotent stem cells into mesenchymal lineage for therapeutic use. Cell Cycle 2019 Nov;18(21):2954-2971.
    doi: 10.1080/15384101.2019.1664224pubmed: 31505996google scholar: lookup
  10. Wang Z, Gong W, Yao Z, Jin K, Niu Y, Li B, Zuo Q. Mechanisms of Embryonic Stem Cell Pluripotency Maintenance and Their Application in Livestock and Poultry Breeding. Animals (Basel) 2024 Jun 9;14(12).
    doi: 10.3390/ani14121742pubmed: 38929361google scholar: lookup
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