Isolation and Characterization of Mesenchymal Stem Cells from Human Skin

Main Article Content

Authors

A.B. Aralbek

L. N. Gumilyov Eurasian National University, Satpayev Street, Astana 010000, Kazakhstan

V.B. Ogay

National Center for Biotechnology, Qorgalzhyn Highway 13/5, Astana 010000, Kazakhstan

Abstract

Mesenchymal stem cells (MSCs) derived from human skin dermis are of considerable interest in regenerative medicine due to their high proliferative activity, self-renewal capacity, and multilineage differentiation potential. In this study, a primary culture of dermal MSCs was obtained and characterized through enzymatic digestion of skin biopsies using dispase and collagenase, followed by cultivation in various nutrient media. Morphological assessment, karyotyping, and immunophenotypic analysis confirmed that the cells belonged to the MSC population (CD105⁺, CD90⁺, CD45⁻). Comparative experiments were conducted to identify the optimal culture medium and seeding density. It was established that DMEM/F12 + 10% FBS medium yielded the highest increase in cell mass, while a seeding density of 3.5×10³ cells/cm² supported effective proliferative expansion. Moreover, the dermal MSCs demonstrated the ability to differentiate into osteoblasts, chondrocytes, and adipocytes. The results obtained highlight the high biological activity of human skin MSCs and their promising potential for applications in cell therapy and tissue engineering.

Keywords

Skin-derived mesenchymal stem cells, clonogenicity, immunophenotype, multilineage differentiation, regenerative medicine

Article Details

References

Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells // Science. 1999. – Vol. 284, № 5411. – P. 143–147.

Caplan AI. Mesenchymal stem cells // J Cell Biochem. 1991. – Vol. 47, № 3. – P. 256–264.

Uccelli A, Moretta L, Pistoia V. Mesenchymal stem cells in health and disease // Nat Rev Immunol. 2008. – Vol. 8, № 9. – P. 726–736.

Squillaro T, Peluso G, Galderisi U. Clinical trials with mesenchymal stem cells: An update // Cell Transplant. 2016. – Vol. 25, № 5. – P. 829–848.

Galipeau J, Sensébé L. Mesenchymal stromal cells: Clinical challenges and therapeutic opportunities // Cytotherapy. 2018. – Vol. 22, № 6. – P. 824–833.

Shi Y, Wang Y, Li Q, Liu K, Hou J, Shao C, et al. Immunoregulatory mechanisms of mesenchymal stem and stromal cells in inflammatory diseases // Nat Rev Immunol. 2018. – Vol. 14, № 8. – P. 493–507.

Vizoso FJ, Eiro N, Cid S, Schneider J, Perez-Fernandez R. Mesenchymal stem cell secretome: Toward cell-free therapeutic strategies in regenerative medicine // Int J Mol Sci. 2017. – Vol. 18, № 9. – P. 1852.

Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, et al. Multilineage cells from human adipose tissue: Implications for cell-based therapies // Tissue Eng. 2001. – Vol. 7, № 2. – P. 211–228.

Mason C, Dunnill P. A brief definition of regenerative medicine // Regen Med. 2008. – Vol. 3, № 1. – P. 1–5.

Huang GTJ, Gronthos S, Shi S. Mesenchymal stem cells derived from dental tissues vs. those from other sources: Their biology and role in regenerative medicine // J Dent Res. 2009. – Vol. 88, № 9. – P. 792–806.

Riekstina U, Cakstina I, Parfejevs V, Hoogduijn M, Jankovskis G, Muiznieks I, et al. Characterization of human skin-derived mesenchymal stem cell proliferation rate in vitro // Stem Cells. 2008. – Vol. 26, № 11. – P. 2875–2883.

Toma JG, McKenzie IA, Bagli D, Miller FD. Isolation and Characterization of Multipotent Skin-Derived Precursors from Human Skin // Stem Cells. 2005. – Vol. 23, № 6. – P. 727–737.

Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini FC, Krause DS, et al. Minimal criteria for defining multipotent mesenchymal stromal cells: The International Society for Cellular Therapy position statement // Cytotherapy. 2006. – Vol. 8, № 4. – P. 315–317.

Han S, Kim J, Lee G, Na J, Kang Y. Enhanced viability and functionality of mesenchymal stem cells under hypoxic conditions // Stem Cell Res Ther. 2017. – Vol. 8, № 1. – P. 18.

Kim M, Kim C, Choi YS, Park HS, Lee S, Lee WJ, et al. Maintenance and regulation of stemness in mesenchymal stem cells by low oxygen concentration // Cell Physiol Biochem. 2018. – Vol. 49, № 2. – P. 842–854.

Sensebé L, Bourin P, Tarte K. Good manufacturing practices for mesenchymal stromal cells: Issues and challenges // Cytotherapy. 2013. – Vol. 15, № 4. – P. 362–366.

Baxter MA, Wynn RF, Jowitt SN, Wraith JE, Fairbairn LJ, Bellantuono I. Study of telomere length reveals rapid aging of human marrow stromal cells following in vitro expansion // Stem Cells. 2004. – Vol. 22, № 5. – P. 675–682.

Lindroos B, Suuronen R, Miettinen S. The potential of adipose stem cells in regenerative medicine // Stem Cell Rev Rep. 2011. – Vol. 4, № 2. – P. 269–291.

Mohammadi S, Nikbakht M, Tavakolifar Y, Gholami M, Taheri M, Vahidinia A, et al. Optimization of culture conditions for human mesenchymal stem cells using different media and seeding densities // Stem Cell Res Ther. 2016. – Vol. 7, № 1. – P. 97.