Skip to main content

Isolation and Primary Culture Methods of Adult and Larval Myogenic Cells from Xenopus laevis

  • Protocol
  • First Online:
Xenopus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1865))

Abstract

During amphibian metamorphosis, larval-to-adult conversion of the myogenic system occurs and there are two distinct types of muscle stem cells; larval myogenic cells have a death-fate by apoptosis in the presence of thyroid hormone T3, and adult myogenic cells have a life-fate under the same conditions. Here, we describe isolation and culture methods for adult and larval myogenic cells from the frog, Xenopus laevis. Both types of cultured myogenic cells undergo cell division and cell differentiation, i.e., formation of multinucleated myotubes in serum-containing medium. Insulin-like growth factor-1 promotes cell division and differentiation in both cells. Basic properties and some applications of primary culture are also described.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Shibota Y, Kaneko Y, Kuroda M et al (2000) Larval-to-adult conversion of a myogenic system in the frog, Xenopus laevis, by larval-type myoblast-specific control of cell division, cell differentiation, and programmed cell death by triiodo-L-thyronine. Differentiation 66:227–238

    Article  CAS  Google Scholar 

  2. Nishikawa A, Hayashi H (1994) Isoform transition of contractile proteins related to muscle remodeling with an axial gradient during metamorphosis in Xenopus laevis. Dev Biol 165:86–94

    Article  CAS  PubMed  Google Scholar 

  3. Nishikawa A, Hayashi H (1995) Spatial, temporal and hormonal regulation of programmed muscle cell death during metamorphosis of the frog Xenopus laevis. Differentiation 59:207–214

    Article  CAS  Google Scholar 

  4. Nishikawa A, Murata E, Akita M et al (1998) Roles of macrophages in programmed cell death and remodeling of tail and body muscle of Xenopus laevis during metamorphosis. Histochem Cell Biol 109:11–17

    Article  CAS  Google Scholar 

  5. Soriano-Arroquia A, Clegg PD, Molloy AP et al (2017) Preparation and culture of myogenic precursor cells/primary myoblasts from skeletal muscle of adult and aged humans. J Vis Exp (120):55047. https://doi.org/10.3791/55047

  6. Danoviz ME Yablonka-Reuveni Z (2012) Skeletal muscle satellite cells: background and methods for isolation and analysis in a primary culture system. Methods Mol Biol 798:21–52

    Article  Google Scholar 

  7. Schmid C, Steiner T, Froesch ER (1983) Preferential enhancement of myoblast differentiation by insulin-like growth factors (IGF I and IGF II) in primary cultures of chicken embryonic cells. FEBS Lett 161:117–121

    Article  CAS  Google Scholar 

  8. Froehlich JM, Seiliez I, Gabillard J-C, et al (2014) Preparation of primary myogenic precursor cell/myoblast cultures from basal vertebrate lineages. J Vis Exp (86):e51354. doi:https://doi.org/10.3791/51354

  9. Gabillard JC, Sabin N, Paboeuf G (2010) In vitro characterization of proliferation and differentiation of trout satellite cells. Cell Tissue Res 342:471–477

    Article  Google Scholar 

  10. Mulvaney DR, Cyrino JEP (1995) Establishment of channel catfish satellite cell cultures. Bas Appl Myo 5:65–70

    Google Scholar 

  11. Shimizu-Nishikawa K, Shibota Y, Takei A et al (2002) Regulation of specific developmental fates of larval- and adult-type muscles during metamorphosis of the frog Xenopus. Dev Biol 251:91–104

    Article  CAS  Google Scholar 

  12. Miyata S, Yada T, Ishikawa N et al (2017) Insulin-like growth factor 1 regulation of proliferation and differentiation of Xenopus laevis myogenic cells in vitro. In Vitro Cell Dev Biol-Anim 53:231–247

    Article  CAS  Google Scholar 

  13. Samuels HH, Stanley F, Casanova J (1979) Depletion of L-3, 5, 3′-triiodothyronine and L-thyroxine in euthyroid calf serum for use in cell culture studies of the action of thyroid hormone. Endocrinol 105:80–85

    Article  CAS  Google Scholar 

  14. Nieuwkoop PD, Faber J (1967) Normal table of Xenopus laevis (Daudin). North-Holland Publisher, Amsterdam

    Google Scholar 

Download references

Acknowledgment

This work was supported by a Shimane University Strategic Grant for Innovative Research (Functional Healthy Rice Project).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Akio Nishikawa .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Taheruzzaman, K., Nishikawa, A. (2018). Isolation and Primary Culture Methods of Adult and Larval Myogenic Cells from Xenopus laevis. In: Vleminckx, K. (eds) Xenopus. Methods in Molecular Biology, vol 1865. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8784-9_21

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8784-9_21

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8783-2

  • Online ISBN: 978-1-4939-8784-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics