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The efficiency of in vitro isolation and myogenic differentiation of MSCs derived from adipose connective tissue, bone marrow, and skeletal muscle tissue

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Abstract

The objective of the study is to evaluate efficiency of in vitro isolation and myogenic differentiation of mesenchymal stem cells (MSCs) derived from adipose connective tissue (AD-MSCs), bone marrow (BM-MSCs), and skeletal muscle tissue (MC-MSCs). MSCs were isolated from adipose connective tissue, bone marrow, and skeletal muscle tissue of two adult 6-wk-old rats. Cultured MSCs were treated with 5-azacytidine (AZA) to induce myogenic differentiation. Isolated MSCs and differentiated cells were evaluated by immunocytochemistry (ICC), fluorescence-activated cell sorting (FACS), PCR, and RT-PCR. AD-MSCs showed the highest proliferation rate while BM-MSCs had the lowest one. In ICC, isolated MSCs had strong CD90- and CD44-positive expression and negative expression of CD45, CD31, and CD34, while AZA-treated MSCs had strong positive desmin expression. In FACS analysis, AD-MSCs had the highest percentage of CD90- and CD44-positive-expressing cells (99% and 96%) followed by BM-MSCs (97% and 94%) and MC-MSCs (92% and 91%).At 1 wk after incubation with AZA treatment, the peak of myogenin expression reached 93% in differentiated MC-MSCs, 83.3% in BM-MSCs, and 77% in AD-MSCs. MSCs isolated from adipose connective tissue, bone marrow, and skeletal muscle tissue have the same morphology and phenotype, but AD-MSCs were the most easily accessible and had the highest rate of growth on cultivation and the highest percentage of stem cell marker expression. Moreover, although MC-MSCs showed the highest rate of myogenic differentiation potential and expression of myoblast markers, AD-MSCs and BM-MSCs still can be valuable alternatives. The differentiated myoblastic cells could be an available new choice for myoblastic auto-transplantation in regeneration medicine.

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Authors and Affiliations

  1. Division of Infectious Disease Control, Center for Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan

    Fatma Y. Meligy, Masato Kawabata & Toshiro Shirakawa

  2. Department of Histology, Faculty of Medicine, Assiut University, Assiut, Egypt

    Fatma Y. Meligy

  3. Division of Urology, Department of Organs Therapeutics, Kobe University Graduate School of Medicine, Kobe, Japan

    Katsumi Shigemura, Hosny M. Behnsawy, Masato Fujisawa & Toshiro Shirakawa

  4. Department of Urology, Faculty of Medicine, Assiut University, Assiut, Egypt

    Hosny M. Behnsawy

  5. Division of Infectious Disease Control, Kobe University Graduate School of Health Sciences, Kobe, Japan

    Toshiro Shirakawa

Authors
  1. Fatma Y. Meligy
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  2. Katsumi Shigemura
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  3. Hosny M. Behnsawy
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  4. Masato Fujisawa
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  5. Masato Kawabata
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  6. Toshiro Shirakawa
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Correspondence to Toshiro Shirakawa.

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Editor: T. Okamoto

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Meligy, F.Y., Shigemura, K., Behnsawy, H.M. et al. The efficiency of in vitro isolation and myogenic differentiation of MSCs derived from adipose connective tissue, bone marrow, and skeletal muscle tissue. In Vitro Cell.Dev.Biol.-Animal 48, 203–215 (2012). https://doi.org/10.1007/s11626-012-9488-x

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  • DOI: https://doi.org/10.1007/s11626-012-9488-x

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