Open AccessCharacterization of smooth muscle differentiation of purified human skeletal muscle‐derived cells
Author(s) -
Lu ShingHwa,
Lin Alex T.L.,
Chen KuangKuo,
Chiang Han Sun,
Chang Luke S.
Publication year - 2011
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/j.1582-4934.2010.01017.x
Subject(s) - caldesmon , calponin , myosin , immunofluorescence , biology , microbiology and biotechnology , western blot , myocyte , cellular differentiation , actin , skeletal muscle , gene expression , smooth muscle tissue , cell culture , antibody , gene , smooth muscle , immunology , endocrinology , biochemistry , genetics , calmodulin , enzyme
The purpose of this study is to characterize the smooth muscle differentiation of purified human muscle‐derived cells (hMDCs). The isolation and purification of hMDCs were conducted by modified preplate technique and Dynal CD34 cell selection. Smooth muscle cell differentiation was induced by the use of smooth muscle induction medium (SMIM) and low‐serum medium. The gene expressions at the mRNA and protein levels of undifferentiated and differentiated hMDCs were tested by RT‐PCR, Western blot and immunofluorescence studies. Western blot and immunofluorescence studies demonstrated the purified hMDCs cultured in SMIM for 4 weeks and expressed significant amount of smooth muscle myosin heavy chain (MHC) and α‐smooth muscle actin (ASMA). The cells cultured in low‐serum medium for 4 weeks also expressed ASMA, while the control group did not. RT‐PCR analysis showed increased gene expression of smooth muscle markers, such as ASMA, Calponin, SM22, Caldesmon, Smoothelin and MHC when purified hMDCs were exposed to SMIM for 2 and 4 weeks when compared to the controls. In conclusion, we confirmed the smooth muscle differentiation capability of purified hMDCs. The gene expression of smooth muscle differentiation of purified hMDCs was characterized. These cells may be potential biomaterials for human tissue regeneration.