Skip to main content
SpringerLink
Log in

Isolation and Characterization of Different Mesenchymal Stem Cell Populations from Rat Femur

  • Protocol
  • First Online:
Pre-Clinical Models

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1916))

Abstract

Purified mesenchymal stem cells (MSCs) may be used for a multitude of applications, from the study of biological processes such as cell division and coordinated gene expression to tissue engineering and regenerative medicine. However, although highly similar, MSCs isolated and purified from different tissues may be biologically different in the ability of the cells to respond to environmental cues that instigate and propagate changes in cell fate such as differentiation, proliferation, apoptosis, and senescence. Selecting which MSC subtype to study may therefore profoundly influence the outcome of the investigation. Here we outline the isolation, purification, and differentiation of three different MSC subtypes derived from various depots within rat bone. These include MSCs from bone marrow, compact bone, and the proximal femur. Osteoblastic and adipogenic differentiation exemplify differences between these cells.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 169.99
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. Conget PA, Minguell JJ (1999) Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells. J Cell Physiol 181:67–73. https://doi.org/10.1002/(SICI)1097-4652(199910)181:1<67:AID-JCP7>3.0.CO;2-C

    Article  CAS  PubMed  Google Scholar 

  2. Veronesi F, Torricelli P, Borsari V, Tschon M, Rimondini L, Fini M (2011) Mesenchymal stem cells in the aging and osteoporotic population. Crit Rev Eukaryot Gene Expr 21:363–377

    Article  CAS  PubMed  Google Scholar 

  3. Caplan AI (1991) Mesenchymal stem cells. J Orthop Res Off Publ Orthop Res Soc 9:641–650. https://doi.org/10.1002/jor.1100090504

    Article  CAS  Google Scholar 

  4. Kopen GC, Prockop DJ, Phinney DG (1999) Marrow stromal cells migrate throughout forebrain and cerebellum, and they differentiate into astrocytes after injection into neonatal mouse brains. Proc Natl Acad Sci U S A 96:10711–10716

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD et al (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147

    Article  CAS  PubMed  Google Scholar 

  6. Reger RL, Tucker AH, Wolfe MR (2008) Differentiation and characterization of human MSCs. Methods Mol Biol 449:93–107. https://doi.org/10.1007/978-1-60327-169-1_7

    Article  CAS  PubMed  Google Scholar 

  7. Shi S, Zhang M, Guo R, Miao Y, Zhang X, Li B (2015) Molecular imaging to monitor repair of myocardial infarction using genetically engineered bone marrow-derived mesenchymal stem cells. Curr Gene Ther 15:460–471

    Article  CAS  PubMed  Google Scholar 

  8. Lo Furno D, Mannino G, Giuffrida R (2018) Functional role of mesenchymal stem cells in the treatment of chronic neurodegenerative diseases. J Cell Physiol 233:3982–3999. https://doi.org/10.1002/jcp.26192

    Article  CAS  PubMed  Google Scholar 

  9. Mattar P, Bieback K (2015) Comparing the immunomodulatory properties of bone marrow, adipose tissue, and birth-associated tissue mesenchymal stromal cells. Front Immunol 6:560. https://doi.org/10.3389/fimmu.2015.00560

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Yorukoglu AC, Kiter AE, Akkaya S, Satiroglu-Tufan NL, Tufan AC (2017) A concise review on the use of mesenchymal stem cells in cell sheet-based tissue engineering with special emphasis on bone tissue regeneration. Stem Cells Int 2017:2374161. https://doi.org/10.1155/2017/2374161

    Article  PubMed  PubMed Central  Google Scholar 

  11. Veronesi F, Borsari V, Sartori M, Orciani M, Mattioli-Belmonte M, Fini M (2018) The use of cell conditioned medium for musculoskeletal tissue regeneration. J Cell Physiol 233:4423–4442. https://doi.org/10.1002/jcp.26291

    Article  CAS  PubMed  Google Scholar 

  12. Giebel B, Kordelas L, Börger V (2017) Clinical potential of mesenchymal stem/stromal cell-derived extracellular vesicles. Stem Cell Investig 4:84. https://doi.org/10.21037/sci.2017.09.06

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Zhang B, Yeo RWY, Tan KH, Lim SK (2016) Focus on extracellular vesicles: therapeutic potential of stem cell-derived extracellular vesicles. Int J Mol Sci 17:174. https://doi.org/10.3390/ijms17020174

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Marfy-Smith SJ, Clarkin CE (2017) Are mesenchymal stem cells so bloody great after all? Stem Cells Transl Med 6:3–6. https://doi.org/10.5966/sctm.2016-0026

    Article  PubMed  Google Scholar 

  15. Hong HS, Kim YH, Son Y (2012) Perspectives on mesenchymal stem cells: tissue repair, immune modulation, and tumor homing. Arch Pharm Res 35:201–211. https://doi.org/10.1007/s12272-012-0201-0

    Article  CAS  PubMed  Google Scholar 

  16. Ridge SM, Sullivan FJ, Glynn SA (2017) Mesenchymal stem cells: key players in cancer progression. Mol Cancer 16:31. https://doi.org/10.1186/s12943-017-0597-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Ye H, Cheng J, Tang Y, Liu Z, Xu C, Liu Y et al (2012) Human bone marrow-derived mesenchymal stem cells produced TGFbeta contributes to progression and metastasis of prostate cancer. Cancer Investig 30:513–518. https://doi.org/10.3109/07357907.2012.692171

    Article  CAS  Google Scholar 

  18. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D et al (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8:315–317. https://doi.org/10.1080/14653240600855905

    Article  CAS  PubMed  Google Scholar 

  19. Jaiswal N, Haynesworth SE, Caplan AI, Bruder SP (1997) Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J Cell Biochem 64:295–312

    Article  CAS  PubMed  Google Scholar 

  20. Zhu H, Guo Z-K, Jiang X-X, Li H, Wang XY, Yao HY et al (2010) A protocol for isolation and culture of mesenchymal stem cells from mouse compact bone. Nat Protoc 5:550–560. https://doi.org/10.1038/nprot.2009.238

    Article  CAS  PubMed  Google Scholar 

  21. Maecker HT, Trotter J (2006) Flow cytometry controls, instrument setup, and the determination of positivity. Cytometry A 69:1037–1042. https://doi.org/10.1002/cyto.a.20333

    Article  PubMed  Google Scholar 

  22. Roederer M (2001) Spectral compensation for flow cytometry: visualization artifacts, limitations, and caveats. Cytometry 45:194–205

    Article  CAS  PubMed  Google Scholar 

  23. Gregory CA, Gunn WG, Peister A, Prockop DJ (2004) An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. Anal Biochem 329:77–84. https://doi.org/10.1016/j.ab.2004.02.002

    Article  CAS  PubMed  Google Scholar 

  24. Ogawa R, Mizuno H, Watanabe A, Migita M, Hyakusoku H, Shimada T (2004) Adipogenic differentiation by adipose-derived stem cells harvested from GFP transgenic mice-including relationship of sex differences. Biochem Biophys Res Commun 319:511–517. https://doi.org/10.1016/j.bbrc.2004.05.021

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Endocrinology, Department of Medicine, Faculty of Medicine and Health Sciences, Stellenbosch University, Cape Town, South Africa

    Frans Alexander Jacobs, Mari van de Vyver & William Frank Ferris

Authors
  1. Frans Alexander Jacobs
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mari van de Vyver
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. William Frank Ferris
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to William Frank Ferris .

Editor information

Editors and Affiliations

  1. Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Campinas, São Paulo, Brazil

    Paul C. Guest

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Jacobs, F.A., van de Vyver, M., Ferris, W.F. (2019). Isolation and Characterization of Different Mesenchymal Stem Cell Populations from Rat Femur. In: Guest, P. (eds) Pre-Clinical Models. Methods in Molecular Biology, vol 1916. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8994-2_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8994-2_13

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8993-5

  • Online ISBN: 978-1-4939-8994-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation