Advertisement

Cytotechnology

, Volume 70, Issue 6, pp 1671–1683 | Cite as

A modified preplate technique for efficient isolation and proliferation of mice muscle-derived stem cells

  • Zhuqiu Xu
  • Lu Yu
  • Haibin Lu
  • Weifeng Feng
  • Lulu Chen
  • Jing Zhou
  • Xiaonan YangEmail author
  • Zuoliang QiEmail author
Original Article
  • 195 Downloads

Abstract

We modified an existing protocol to develop a more efficient method to acquire and culture muscle-derived stem cells (MDSCs) and compared the characteristics of cells obtained from the two methods. This method is based on currently used multistep enzymatic digestion and preplate technique. During the replating process, we replaced the traditional medium with isolation medium to promote fibroblast-like cell adherence at initial replating step, which shortened the purifying duration by up to 4 days. Moreover, we modified the culture container to provide a stable microenvironment that promotes MDSC adherence. We compared the cell morphology, growth curve and the expression of specific markers (Sca-1, CD34, PAX7 and Desmin) between the two cell groups separately obtained from the two methods. Afterwards, we compared the neural differentiation capacity of MDSCs with other muscle-derived cell lineages. The protocol developed here is a fast and effective method to harvest and purify MDSCs from mice limb skeletal muscle.

Keywords

Regenerative medicine Cell and tissue culture Myogenic stem cells In vitro culture 

Notes

Author’s contribution

ZX and LY performed the biopsy and cells culture, HL and WF took charge of the immunofluorescence and flow cytometry, LC and JZ accomplished Statistical analysis. ZX drafted the manuscript. Co-corresponding authors XY and ZQ designed and guided the study. All authors have read and approved the final manuscript.

Funding

This work was supported by the National Natural Science Foundation of China (81571921 and 81671908), the Union Youth Science and Research Fund (3332015155), the Science Fund of Plastic Surgery Hospital, CAMS, PUMC (Q2015013) and the Innovation Fund of Graduate Students, CAMS, PUMC (2017-1002-1-10).

Compliance with ethical standards

Conflict of interest

The authors declare they have no competing interests.

References

  1. Agley CC, Rowlerson AM, Velloso CP, Lazarus NL, Harridge SD (2015) Isolation and quantitative immunocytochemical characterization of primary myogenic cells and fibroblasts from human skeletal muscle. J Vis Exp 95:52049Google Scholar
  2. Arsic N, Mamaeva D, Lamb NJ, Fernandez A (2008) Muscle-derived stem cells isolated as non-adherent population give rise to cardiac, skeletal muscle and neural lineages. Exp Cell Res 314:1266–1280CrossRefGoogle Scholar
  3. Asakura A, Komaki M, Rudnicki M (2001) Muscle satellite cells are multipotential stem cells that exhibit myogenic, osteogenic, and adipogenic differentiation. Differentiation 68:245–253CrossRefGoogle Scholar
  4. Beane OS, Fonseca VC, Cooper LL, Koren G, Darling EM (2014) Impact of aging on the regenerative properties of bone marrow-, muscle-, and adipose-derived mesenchymal stem/stromal cells. PLoS ONE 9:e115963CrossRefGoogle Scholar
  5. Che X, Guo J, Wang B, Bai Y (2011) Rapid isolation of muscle-derived stem cells by discontinuous Percoll density gradient centrifugation. In Vitro Cell Dev Biol Anim 47:454–458CrossRefGoogle Scholar
  6. Chen S, Lewallen M, Xie T (2013) Adhesion in the stem cell niche: biological roles and regulation. Development 140:255–265CrossRefGoogle Scholar
  7. Chen B, Ding J, Zhang W, Zhou G, Cao Y, Liu W, Wang B (2016) Tissue engineering of tendons: a comparison of muscle-derived cells, tenocytes, and dermal fibroblasts as cell sources. Plast Reconstr Surg 137:536e–544eCrossRefGoogle Scholar
  8. Collins CA, Olsen I, Zammit PS, Heslop L, Petrie A, Partridge TA, Morgan JE (2005) Stem cell function, self-renewal, and behavioral heterogeneity of cells from the adult muscle satellite cell niche. Cell 122:289–301CrossRefGoogle Scholar
  9. Cottle BJ, Lewis FC, Shone V, Ellison-Hughes GM (2017) Skeletal muscle-derived interstitial progenitor cells (PICs) display stem cell properties, being clonogenic, self-renewing, and multi-potent in vitro and in vivo. Stem Cell Res Ther 8:158CrossRefGoogle Scholar
  10. Gharaibeh B, Lu A, Tebbets J, Zheng B, Feduska J, Crisan M, Peault B, Cummins J, Huard J (2008) Isolation of a slowly adhering cell fraction containing stem cells from murine skeletal muscle by the preplate technique. Nat Protoc 3:1501–1509CrossRefGoogle Scholar
  11. Gussoni E, Soneoka Y, Strickland CD, Buzney EA, Khan MK, Flint AF, Kunkel LM, Mulligan RC (1999) Dystrophin expression in the mdx mouse restored by stem cell transplantation. Nature 401:390–394PubMedGoogle Scholar
  12. Han D, Chen S, Fang S, Liu S, Jin M, Guo Z, Yuan Y, Wang Y, Liu C, Mei X (2017) The neuroprotective effects of muscle-derived stem cells via brain-derived neurotrophic factor in spinal cord injury model. Biomed Res Int 2017:1972608PubMedPubMedCentralGoogle Scholar
  13. Jackson KA, Mi T, Goodell MA (1999) Hematopoietic potential of stem cells isolated from murine skeletal muscle. Proc Natl Acad Sci USA 96:14482–14486CrossRefGoogle Scholar
  14. Jackson WM, Alexander PG, Bulken-Hoover JD, Vogler JA, Ji Y, McKay P, Nesti LJ, Tuan RS (2013) Mesenchymal progenitor cells derived from traumatized muscle enhance neurite growth. J Tissue Eng Regen Med 7:443–451CrossRefGoogle Scholar
  15. Jankowski RJ, Deasy BM, Huard J (2002) Muscle-derived stem cells. Gene Ther 9:642–647CrossRefGoogle Scholar
  16. Kallestad KM, McLoon LK (2010) Defining the heterogeneity of skeletal muscle-derived side and main population cells isolated immediately ex vivo. J Cell Physiol 222:676–684PubMedPubMedCentralGoogle Scholar
  17. Kalvelyte A, Krestnikova N, Stulpinas A, Bukelskiene V, Bironaite D, Baltriukiene D, Imbrasaite A (2013) Long-term muscle-derived cell culture: multipotency and susceptibility to cell death stimuli. Cell Biol Int 37:292–304CrossRefGoogle Scholar
  18. Kelc R, Trapecar M, Vogrin M, Cencic A (2013) Skeletal muscle-derived cell cultures as potent models in regenerative medicine research. Muscle Nerve 47:477–482CrossRefGoogle Scholar
  19. Khosravi-Farsani S, Amidi F, Habibi Roudkenar M, Sobhani A (2015) Isolation and enrichment of mouse female germ line stem cells. Cell J 16:406–415PubMedPubMedCentralGoogle Scholar
  20. Lau AM, Tseng YH, Schulz TJ (2014) Adipogenic fate commitment of muscle-derived progenitor cells: isolation, culture, and differentiation. Methods Mol Biol 1213:229–243CrossRefGoogle Scholar
  21. Lavasani M, Lu A, Thompson SD, Robbins PD, Huard J, Niedernhofer LJ (2013) Isolation of muscle-derived stem/progenitor cells based on adhesion characteristics to collagen-coated surfaces. Methods Mol Biol 976:53–65CrossRefGoogle Scholar
  22. Lavasani M, Thompson SD, Pollett JB, Usas A, Lu A, Stolz DB, Clark KA, Sun B, Peault B, Huard J (2014) Human muscle-derived stem/progenitor cells promote functional murine peripheral nerve regeneration. J Clin Invest 124:1745–1756CrossRefGoogle Scholar
  23. Lee JY, Qu-Petersen Z, Cao B, Kimura S, Jankowski R, Cummins J, Usas A, Gates C, Robbins P, Wernig A, Huard J (2000) Clonal isolation of muscle-derived cells capable of enhancing muscle regeneration and bone healing. J Cell Biol 150:1085–1100CrossRefGoogle Scholar
  24. Lee JY, Piao S, Kim IG, Byun SS, Hwang JH, Hong SH, Kim SW, Hwang TK, Lee JY (2012) Effect of human muscle-derived stem cells on cryoinjured mouse bladder contractility. Urology 80:224.e7–224.e11Google Scholar
  25. Li Y, Pan H, Huard J (2010) Isolating stem cells from soft musculoskeletal tissues. J Vis Exp.  https://doi.org/10.3791/2011
  26. Li H, Usas A, Poddar M, Chen CW, Thompson S, Ahani B, Cummins J, Lavasani M, Huard J (2013) Platelet-rich plasma promotes the proliferation of human muscle derived progenitor cells and maintains their stemness. PLoS ONE 8:e64923CrossRefGoogle Scholar
  27. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25(4):402–408CrossRefGoogle Scholar
  28. Lorant J, Saury C, Schleder C, Robriquet F, Lieubeau B, Negroni E, Leroux I, Chabrand L, Viau S, Babarit C, Ledevin M, Dubreil L, Hamel A, Magot A, Thorin C, Guevel L, Delorme B, Pereon Y, Butler-Browne G, Mouly V, Rouger K (2018) Skeletal muscle regenerative potential of human mustem cells following transplantation into injured mice muscle. Mol Ther 26:618–633CrossRefGoogle Scholar
  29. Lu A, Proto JD, Guo L, Tang Y, Lavasani M, Tilstra JS, Niedernhofer LJ, Wang B, Guttridge DC, Robbins PD, Huard J (2012) NF-κB negatively impacts the myogenic potential of muscle-derived stem cells. Mol Ther 20:661–668CrossRefGoogle Scholar
  30. Lucas PA, Calcutt AF, Southerland SS, Alan Wilson J, Harvey RL, Warejcka D, Young HE (1995) A population of cells resident within embryonic and newborn rat skeletal muscle is capable of differentiating into multiple mesodermal phenotypes. Wound Repair Regen 3:449–460CrossRefGoogle Scholar
  31. Machida S, Spangenburg EE, Booth FW (2004) Primary rat muscle progenitor cells have decreased proliferation and myotube formation during passages. Cell Prolif 37:267–277CrossRefGoogle Scholar
  32. Meng J, Chun S, Asfahani R, Lochmuller H, Muntoni F, Morgan J (2014) Human skeletal muscle-derived CD133(+) cells form functional satellite cells after intramuscular transplantation in immunodeficient host mice. Mol Ther 22:1008–1017CrossRefGoogle Scholar
  33. Montarras D, Morgan J, Collins C, Relaix F, Zaffran S, Cumano A, Partridge T, Buckingham M (2005) Direct isolation of satellite cells for skeletal muscle regeneration. Science 309:2064–2067CrossRefGoogle Scholar
  34. Morgan JE, Partridge TA (2003) Muscle satellite cells. Int J Biochem Cell Biol 35:1151–1156CrossRefGoogle Scholar
  35. Nakajima N, Tamaki T, Hirata M, Soeda S, Nitta M, Hoshi A, Terachi T (2017) Purified human skeletal muscle-derived stem cells enhance the repair and regeneration in the damaged urethra. Transplantation 101(10):2312–2320CrossRefGoogle Scholar
  36. Ozasa Y, Gingery A, Thoreson AR, An KN, Zhao C, Amadio PC (2014) A comparative study of the effects of growth and differentiation factor 5 on muscle-derived stem cells and bone marrow stromal cells in an in vitro tendon healing model. J Hand Surg Am 39:1706–1713CrossRefGoogle Scholar
  37. Park MH, Park JE, Kim MS, Lee KY, Park HJ, Yun JI, Choi JH, Lee E, Lee ST (2014) Development of a high-yield technique to isolate spermatogonial stem cells from porcine testes. J Assist Reprod Genet 31:983–991CrossRefGoogle Scholar
  38. Qu-Petersen Z, Deasy B, Jankowski R, Ikezawa M, Cummins J, Pruchnic R, Mytinger J, Cao B, Gates C, Wernig A, Huard J (2002) Identification of a novel population of muscle stem cells in mice: potential for muscle regeneration. J Cell Biol 157:851–864CrossRefGoogle Scholar
  39. Saito K, Tamaki T, Hirata M, Hashimoto H, Nakazato K, Nakajima N, Kazuno A, Sakai A, Iida M, Okami K (2015) Reconstruction of multiple facial nerve branches using skeletal muscle-derived multipotent stem cell sheet-pellet transplantation. PLoS ONE 10:e0138371CrossRefGoogle Scholar
  40. Seta H, Maki D, Kazuno A, Yamato I, Nakajima N, Soeda S, Uchiyama Y, Tamaki T (2018) Voluntary exercise positively affects the recovery of long-nerve gap injury following tube-bridging with human skeletal muscle-derived stem cell transplantation. J Clin Med 7:67CrossRefGoogle Scholar
  41. Spradling A, Drummond-Barbosa D, Kai T (2001) Stem cells find their niche. Nature 414:98–104CrossRefGoogle Scholar
  42. Tamaki T, Akatsuka A, Ando K, Nakamura Y, Matsuzawa H, Hotta T, Roy RR, Edgerton VR (2002) Identification of myogenic-endothelial progenitor cells in the interstitial spaces of skeletal muscle. J Cell Biol 157:571–577CrossRefGoogle Scholar
  43. Tamaki T, Uchiyama Y, Okada Y, Ishikawa T, Sato M, Akatsuka A, Asahara T (2005) Functional recovery of damaged skeletal muscle through synchronized vasculogenesis, myogenesis, and neurogenesis by muscle-derived stem cells. Circulation 112:2857–2866CrossRefGoogle Scholar
  44. Tamaki T, Okada Y, Uchiyama Y, Tono K, Masuda M, Nitta M, Hoshi A, Akatsuka A (2008) Skeletal muscle-derived CD34 +/45- and CD34-/45- stem cells are situated hierarchically upstream of Pax7 + cells. Stem Cells Dev 17:653–667CrossRefGoogle Scholar
  45. Tamaki T, Tono K, Uchiyama Y, Okada Y, Masuda M, Soeda S, Nitta M, Akatsuka A (2011) Origin and hierarchy of basal lamina-forming and -non-forming myogenic cells in mouse skeletal muscle in relation to adhesive capacity and Pax7 expression in vitro. Cell Tissue Res 344:147–168CrossRefGoogle Scholar
  46. Tamaki T, Uchiyama Y, Hirata M, Hashimoto H, Nakajima N, Saito K, Terachi T, Mochida J (2015) Therapeutic isolation and expansion of human skeletal muscle-derived stem cells for the use of muscle–nerve–blood vessel reconstitution. Front Physiol 6:165CrossRefGoogle Scholar
  47. Tamaki T, Hirata M, Nakajima N, Saito K, Hashimoto H, Soeda S, Uchiyama Y, Watanabe M (2016) A long-gap peripheral nerve injury therapy using human skeletal muscle-derived stem cells (Sk-SCs): an achievement of significant morphological, numerical and functional recovery. PLoS ONE 11:e0166639CrossRefGoogle Scholar
  48. Vojnits K, Pan H, Dai X, Sun H, Tong Q, Darabi R, Huard J, Li Y (2017) Functional neuronal differentiation of injury-induced muscle-derived stem cell-like cells with therapeutic implications. Sci Rep 7:1177CrossRefGoogle Scholar
  49. Vukusic K, Jonsson M, Brantsing C, Dellgren G, Jeppsson A, Lindahl A, Asp J (2013) High density sphere culture of adult cardiac cells increases the levels of cardiac and progenitor markers and shows signs of vasculogenesis. Biomed Res Int 2013:696837CrossRefGoogle Scholar
  50. Wang HD, Guo Q, Quan A, Lopez J, Alonso-Escalante JC, Lough DM, Lee WPA, Brandacher G, Kumar AR (2017) Vascular endothelial growth factor induction of muscle-derived stem cells enhances vascular phenotype while preserving myogenic potential. Ann Plast Surg 79:404–409CrossRefGoogle Scholar
  51. Wu X, Wang S, Chen B, An X (2010) Muscle-derived stem cells: isolation, characterization, differentiation, and application in cell and gene therapy. Cell Tissue Res 340:549–567CrossRefGoogle Scholar
  52. Xue R, Li JY, Yeh Y, Yang L, Chien S (2013) Effects of matrix elasticity and cell density on human mesenchymal stem cells differentiation. J Orthop Res 31:1360–1365CrossRefGoogle Scholar
  53. Zambon JP, de Sa Barretto LS, Nakamura AN, Duailibi S, Leite K, Magalhaes RS, Orlando G, Ross CL, Peloso A, Almeida FG (2014) Histological changes induced by polyglycolic-acid (PGA) scaffolds seeded with autologous adipose or muscle-derived stem cells when implanted on rabbit bladder. Organogenesis 10:278–288CrossRefGoogle Scholar
  54. Zheng B, Cao B, Crisan M, Sun B, Li G, Logar A, Yap S, Pollett JB, Drowley L, Cassino T, Gharaibeh B, Deasy BM, Huard J, Peault B (2007) Prospective identification of myogenic endothelial cells in human skeletal muscle. Nat Biotechnol 25:1025–1034CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Chinese Academy of Medical SciencePeking Union Medical College, Plastic Surgery HospitalBeijingChina

Personalised recommendations