Abstract
The minimal criteria for mesenchymal stem/stromal cell (MSC) identification set by the International Society for Cellular Therapy include plastic adherence, presence and absence of a set of surface antigens and in vitro multilineage differentiation. This differentiation is assessed through stimulation of MSCs with defined combination and concentration of growth factors towards specific lineages and histological confirmation of the presence of differentiated cells. Here we provide protocols for multilineage differentiation, namely, osteogenesis, adipogenesis, chondrogenesis and myogenesis. We also provide their respective histological analyses.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Kalinina NI, Sysoeva VY, Rubina KA et al (2011) Mesenchymal stem cells in tissue growth and repair. Acta Nat 3:30–37
Qi K, Li N, Zhang Z et al (2018) Tissue regeneration: the crosstalk between mesenchymal stem cells and immune response. Cell Immunol 326:86–93. https://doi.org/10.1016/j.cellimm.2017.11.010
Friedenstein AJ, Gorskaja JF, Kulagina NN (1976) Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 4:267–274
Beane OS, Fonseca VC, Cooper LL et al (2014) Impact of aging on the regenerative properties of bone-marrow-, muscle-, and adipose-derived mesenchymal stem/ stromal cells. PLoS One 9:1–22. https://doi.org/10.1371/journal.pone.0115963
Wagner W, Wein F, Seckinger A et al (2005) Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood. Exp Hematol 33:1402–1416
Dominici M, Le BK, Mueller I et al (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8:315–317
Čamernik K, Barlič A, Drobnič M et al (2018) Mesenchymal stem cells in the musculoskeletal system: from animal models to human tissue regeneration. Stem Cell Rev 14(3):346–369. https://doi.org/10.1007/s12015-018-9800-6
Burlacu A, Rosca AM, Maniu H et al (2008) Promoting effect of 5-azacytidine on the myogenic differentiation of bone marrow stromal cells. Eur J Cell Biol 87:173–184. https://doi.org/10.1016/j.ejcb.2007.09.003
Wakitani S, Saito T, Caplan A (1995) Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine. Muscle Nerve 18:1417–1426
Yang Y-HK, Ogando CR, Wang See C et al (2018) Changes in phenotype and differentiation potential of human mesenchymal stem cells aging in vitro. Stem Cell Res Ther 9:131. https://doi.org/10.1186/s13287-018-0876-3
Langenbach F, Handschel JR (2013) Effects of dexamethasone, ascorbic acid and ß-glycerophosphate on the osteogenic differentiation of stem cells in vitro. Stem Cell Res Ther 4:117. https://doi.org/10.1186/scrt328
Xu N, Liu H, Qu F et al (2013) Hypoxia inhibits the differentiation of mesenchymal stem cells into osteoblasts by activation of Notch signaling. Exp Mol Pathol 94:33–39. https://doi.org/10.1016/j.yexmp.2012.08.003
Autengruber A, Gereke M, Hansen G et al (2012) Impact of enzymatic tissue disintegration on the level of surface molecule expression and immune cell function. Eur J Microbiol Immunol 2:112–120. https://doi.org/10.1556/EuJMI.2.2012.2.3
Zuliani CC, Bombini MF, de AKC et al (2018) Micromass cultures are effective for differentiation of human amniotic fluid stem cells into chondrocytes. Clinics (Sao Paulo) 73:e268. https://doi.org/10.6061/clinics/2018/e268
Ruedel A, Hofmeister S, Bosserhoff A-K (2013) Development of a model system to analyze chondrogenic differentiation of mesenchymal stem cells. Int J Clin Exp Pathol 6:3042–3048
Acknowledgements
This work was supported by the Slovenian Research Agency, J3-7245 Research Project and P3-0298 Research Programme.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media New York
About this protocol
Cite this protocol
Čamernik, K., Zupan, J. (2018). Complete Assessment of Multilineage Differentiation Potential of Human Skeletal Muscle-Derived Mesenchymal Stem/Stromal Cells. In: Turksen, K. (eds) Stem Cells and Aging . Methods in Molecular Biology, vol 2045. Humana, New York, NY. https://doi.org/10.1007/7651_2018_200
Download citation
DOI: https://doi.org/10.1007/7651_2018_200
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9712-1
Online ISBN: 978-1-4939-9713-8
eBook Packages: Springer Protocols