Cardiomyocyte-like differentiation of bone marrow mesenchymal stem cells: an association between related gene expression and cell morphological changes
10.3969/j.issn.2095-4344.2017.13.002
- VernacularTitle:骨髓间充质干细胞分化为心肌样细胞相关基因表达与形态结构发生的关联
- Author:
Yichen XU
;
Linglong LIU
;
Wenjing ZHAO
;
Huifeng WANG
;
Yashu WEI
;
Weiping CHEN
- From:
Chinese Journal of Tissue Engineering Research
2017;21(13):1974-1979
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Bone marrow mesenchymal stem cells (BMSCs) have been induced to differentiate into cardiomyocyte-like cells in vitro.OBJECTIVE:To explore the association between GATA-4, Nkx-2.5 and α-myosin heavy chain (α-MHC) expression and cell morphological changes and structure formation in the process of BMSCs differentiation into cardiomyocyte-like cells.METHODS:By using myocardial lysate, BMSCs were induced to differentiate into cardiomyocytes.Immunocytochemistry staining was used to detect cardiac troponin T (cTnT) and connexin43, for the identification of cardiomyocytes. In the process of directional differentiation, RT-PCR was used to detect the expression of GATA-4,Nkx2.5 and α-MHC.RESULTS AND CONCLUSION:During the directional differentiation of BMSCs, the cells were changed from long fusiform to short rod, forming protrusions that were interconnected to form mesh-like, bamboo-like or myotube-like structure. When the cells were interconnected like a bamboo, cTNT and connexin43 positive cells were visible, and then the number of positive cells increased with the presence of myotube-like structure. RT-PCR results showed that during the induced directional differentiation of BMSCs, GATA-4, Nkx2.5 and α-MHC mRNA levels increased continuously. When interconnected cells formed a mesh-like structure, GATA-4 expression reached the peak and then kept a high level. When adjacent cells were fused into a myotube-like structure, α-MHC reached the peak. Additionally, the expression of Nkx2.5presented a time-dependent increase trend. Overall, during the induced differentiation of BMSCs into cardiomyocyte-like cells, the expression of cardiomyocyte specific genes, characterized by temporality and spatiality, is related to the changes of cell morphology and special structure formation.
