Notch1 is associated with the differentiation of human bone marrow‑derived mesenchymal stem cells to cardiomyocytes
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Updated: Mar 13, 2026

Generation and Expansion of Human Cardiomyocytes from Patient Peripheral Blood Mononuclear Cells
Published on: February 12, 2021
Zipu Yu1, Yu Zou1, Jingya Fan1
1Department of Cardiac Surgery, First Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang 310000, P.R. China.
Insights
The Notch1 signaling pathway may drive the differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) into cardiomyocytes. This process, induced by 5-azacytidine, involves the upregulation of key cardiac transcription factors.
Area of Science:
- Stem cell biology
- Cardiovascular research
- Cell signaling
Background:
- Notch signaling is crucial for stem cell fate determination.
- The specific role of Notch in human bone marrow-derived mesenchymal stem cells (hBMSCs) differentiation into cardiomyocytes is not fully understood.
Purpose of the Study:
- To investigate the involvement of Notch signaling in the in vitro differentiation of hBMSCs into cardiomyocytes.
- To elucidate the molecular mechanisms underlying this differentiation process.
Main Methods:
- Human bone marrow-derived mesenchymal stem cells (hBMSCs) were isolated and characterized using flow cytometry.
- hBMSCs were induced to differentiate into cardiomyocytes using 5-azacytidine (5-aza).
- Differentiation was assessed via morphological changes, western blotting, immunocytochemistry, RT-qPCR, and transmission electron microscopy.
Main Results:
- 5-azacytidine successfully induced hBMSC differentiation into cardiomyocytes, confirmed by increased expression of cardiac markers like α-actin and cardiac troponin T.
- Early upregulation of Notch1 mRNA was observed during differentiation.
- Increased expression of cardiac transcription factors GATA binding protein-4 (GATA4) and NK2 homeobox 5 (Nkx2.5) was detected at later stages.
Conclusions:
- In vitro differentiation of hBMSCs into cardiomyocytes can be achieved using 5-azacytidine.
- The Notch1 signaling pathway appears to play a significant role in this differentiation process.
- Upregulation of Notch1, GATA4, and Nkx2.5 is associated with 5-aza-induced hBMSC cardiomyogenesis.
Abstract:
Notch signaling is involved in the early process of differentiation to determine the fate of stem cells. However, the precise role of Notch in human bone marrow‑derived mesenchymal stem cells (hBMSCs) remains unclear. The present study aimed to investigate the involvement of Notch signalling during the course of hBMSC differentiation into cardiomyocytes using hBMSCs, with multilineage differentiation ability, isolated and purified from human bone marrow. Flow cytometric analysis revealed that CD29, CD44 and CD90 were highly expressed on the surface of cells in their fifth passage, whereas detection of CD34, CD45, CD54 and HLA‑DR was negative. Visualization of morphological changes, western blotting, immunocytochemistry and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) demonstrated that hBMSCs differentiate into cardiomyocytes through treatment with 5‑azacytidine (5‑aza). Transmission electron microscopy revealed ultramicroscopic details of differentiated hBMSCs. Western blotting and immunocytochemistry demonstrated increased protein expression levels of α‑actin and cardiac troponin T expression, and RT‑qPCR revealed increased mRNA expression of Notch1 early in the process of differentiation (days 1, 4 and 7), and increased mRNA expression levels of the transcription factors GATA binding protein‑4 and NK2 homeobox 5 at day 28 day. In conclusion, differentiation of hBMSCs into cardiomyocytes was induced in vitro by 5‑aza, and was associated with upregulation of Notch1, GATA binding protein‑4 and Nkx2.5 expression. Overexpression of the Notch1 signaling pathway may represent a potential mechanism underlying the differentiation of hBMSCs.
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