Differentiation of mesenchymal stem cells from human amniotic fluid to cardiomyocyte-like cells

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.

Differentiation of cardiomyocyte-like cells from human amniotic fluid mesenchymal stem cells by combined induction with human platelet lysate and 5-azacytidine

“…The hAF-MSCs obtained from the second-trimester of a pregnancy possess a self-renewal capacity and multilineage differentiation potential [ 18 , 19 , 20 ]. They can differentiate into chondrocytes [ 21 , 22 ], endothelial cells [ 23 ], osteocytes [ 24 ] and cardiomyocytes [ 25 ]. Furthermore, they have been shown to be effective in the treatment of many diseases such as MI [ 26 ].…”

Comparison of the Cardiomyogenic Potency of Human Amniotic Fluid and Bone Marrow Mesenchymal Stem Cells

“…Thus our data collectively suggests that AF-MSC have superior cardiomyogenic potency as compared to BM-MSC. Although, there is no previous report comparing the cardiomyogenic potential of human AF-MSC and BM-MSC, but several studies have also reported a limited cardiomyogenic potential of BM-MSC (6).We have previously shown that BM-MSC exhibit clonal heterogeneity and only 1 out of 4 clones possessed cardiomyogenic potential, highlighting a low frequency of car-diomyogenic cells in BM-MSC (17). It has also been previously shown that AF-MSC have superior neural and hepatic potency in comparison to BM-MSC (13,14).…”

The effects of human platelet lysate versus commercial endothelial growth medium on the endothelial differentiation potential of human amniotic fluid mesenchymal stem cells

“…The potency of hAF-MSCs was confirmed by recent studies that isolated MSCs from the same source. The results showed the differentiation potential of MSCs into chondrocyte, cardiomyocyte and osteoblast [ 44 , 45 , 46 ].…”