Department of Anatomy, Faculty of Medicine, Chiang Mai University, Maharaj Nakorn Chiang Mai Hospital, Chiang Mai 50200, Thailand.
Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Maharaj Nakorn Chiang Mai Hospital, Chiang Mai 50200, Thailand.
Mol Med Rep. 2017 Nov;16(5):6068-6076. doi: 10.3892/mmr.2017.7333. Epub 2017 Aug 23.
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.
缺血性心脏病（IHD）是影响全球死亡率的主要因素。此外，IHD 已成为泰国人口的一个重大健康问题。使用间充质干细胞（MSCs）的干细胞疗法是一种替代治疗方法，已应用于改善患者的生活质量。羊水（AF）含有异质细胞群体，包括 MSC，它是多能干细胞，能够分化为间充质谱系。本研究的目的是评估人（h）AF 的 MSC 特征，并确定其向心肌分化的潜能。通过流式细胞术分析对 MSC 进行特征描述，结果显示细胞表达 MSC 标志物，如 CD44、CD90、人类白细胞抗原-ABC 和 CD73。alamar blue 检测结果表明，细胞增殖率从早期培养阶段持续增加，在细胞培养的第 1 至 5 天增加了 5 倍。在第 17 天观察到最高的细胞增殖率，与第 1 天相比增加了 30 倍。在诱导心肌生成阶段，在第 0 天和第 21 天之间观察到形态变化，结果表明，在诱导心肌生成组中，hAF 衍生的 MSC 在培养 7 天后呈现出肌管样形态。在诱导心肌生成后，在第 21 天进行免疫组织化学染色，在第 7 天和第 21 天进行逆转录-定量聚合酶链反应。这些步骤是为了检测心脏特异性蛋白（GATA4、心肌肌钙蛋白 T、Nkx2.5 和 Connexin43）的蛋白和基因表达水平。本研究结果表明，hAF-MSC 具有分化为心肌细胞样细胞的潜力。因此，可以得出结论，hAF 可能是干细胞治疗和组织工程的合适 MSC 来源。
