The efficiency of in vitro isolation and myogenic differentiation of MSCs derived from adipose connective tissue,bone marrow,and skeletal muscle tissue |
| |
Authors: | Fatma Y Meligy Katsumi Shigemura Hosny M Behnsawy Masato Fujisawa Masato Kawabata Toshiro Shirakawa |
| |
Institution: | (1) Division of Infectious Disease Control, Center for Infectious Diseases, Kobe University Graduate School of Medicine, Kobe, Japan;(2) Department of Histology, Faculty of Medicine, Assiut University, Assiut, Egypt;(3) Division of Urology, Department of Organs Therapeutics, Kobe University Graduate School of Medicine, Kobe, Japan;(4) Department of Urology, Faculty of Medicine, Assiut University, Assiut, Egypt;(5) Division of Infectious Disease Control, Kobe University Graduate School of Health Sciences, Kobe, Japan; |
| |
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. |
| |
Keywords: | |
本文献已被 PubMed SpringerLink 等数据库收录! |
|