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In Vitro Differentiation of Human Mesenchymal Stem Cells into Functional Cardiomyocyte-like Cells
Published on: August 9, 2017
Differentiation of mesenchymal stem cells from human amniotic fluid to cardiomyocyte‑like cells
Runchana Markmee1, Sirinda Aungsuchawan1, Suteera Narakornsak1
1Department of Anatomy, Faculty of Medicine, Chiang Mai University, Maharaj Nakorn Chiang Mai Hospital, Chiang Mai 50200, Thailand.
Insights
Human amniotic fluid mesenchymal stem cells (hAF-MSCs) show potential for treating ischemic heart disease. These cells effectively differentiate into cardiomyocyte-like cells, offering a promising source for regenerative medicine and tissue engineering applications.
Area of Science:
- Regenerative Medicine
- Cardiovascular Research
- Stem Cell Biology
Background:
- Ischemic heart disease (IHD) is a leading cause of global mortality and a significant health concern in Thailand.
- Mesenchymal stem cells (MSCs) derived from amniotic fluid (AF) offer a potential therapeutic avenue for IHD due to their multipotent differentiation capabilities.
- Human amniotic fluid (hAF) contains a diverse cell population, including MSCs with potential for therapeutic applications.
Purpose of the Study:
- To characterize mesenchymal stem cells (MSCs) from human amniotic fluid (hAF).
- To evaluate the cardiogenic differentiation potential of hAF-derived MSCs.
- To assess the suitability of hAF-MSCs for stem cell therapy and tissue engineering.
Main Methods:
- Flow cytometry was used to analyze MSC surface markers (CD44, CD90, HLA-ABC, CD73).
- Alamar blue assay measured cell proliferation rates over 21 days.
- Cardiogenic induction was performed, followed by morphological assessment, immunohistochemistry, and RT-qPCR to detect cardiac-specific gene and protein expression (GATA4, troponin T, Nkx2.5, Connexin43).
Main Results:
- hAF-MSCs expressed characteristic MSC markers and demonstrated significant proliferation.
- Morphological changes and expression of myotube-like structures were observed during cardiogenic induction.
- Key cardiac-specific genes and proteins (GATA4, troponin T, Nkx2.5, Connexin43) were detected, confirming differentiation into cardiomyocyte-like cells.
Conclusions:
- Human amniotic fluid is a viable source of MSCs with robust proliferation capacity.
- hAF-MSCs possess the potential to differentiate into cardiomyocyte-like cells.
- hAF-derived MSCs represent a promising candidate for stem cell therapy and tissue engineering in cardiovascular applications.
Abstract:
Ischemic heart disease (IHD) is a major factor influencing worldwide mortality rates. Furthermore, IHD has become a significant health problem among the Thai population. Stem cell therapy using mesenchymal stem cells (MSCs) is an alternative therapeutic method that has been applied to improve the quality of life of patients. Amniotic fluid (AF) contains a heterogeneous cell population, including MSCs, which are multipotent stem cells that have the capability to differentiate into mesenchymal lineages. The purpose of the present study was to evaluate the MSC characteristics of human (h)AF and determine its potency regarding cardiogenic differentiation. MSC characterization following flow cytometric analysis revealed that the cells expressed MSC markers, cluster of differentiation (CD)44, CD90, human leukocyte antigen‑ABC and CD73. The results of the alamar blue assay demonstrated that cell proliferation rate continuously increased from the early cultivation phase up to 5‑fold during days 1 to 5 of cell culturing. The highest rate of cell proliferation was observed on day 17 with a 30‑fold increase compared with that on day 1. During the cardiogenic induction stage, morphological changes were observed between day 0 and day 21, and it was revealed that the hAF derived‑MSCs in the cardiogenic‑induced group exhibited myotube‑like morphology after 7 days of cell culturing. Following cardiogenic induction, immunohistochemistry staining was performed on day 21, and reverse transcription‑quantitative polymerase chain reaction on day 7 and 21. These steps were performed to detect the protein and gene expression levels of cardiac specific proteins (GATA4, cardiac troponin T, Nkx2.5 and Connexin43). The results of the present study indicated that hAF‑MSCs possess the potential to differentiate into cardiomyocyte‑like cells. Thus, it was concluded that hAF may be a suitable source of MSCs for stem cell therapy and tissue engineering.
