Dalian R&D Center for Stem Cell and Tissue Engineering, State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian, 116024, China.
Appl Biochem Biotechnol. 2013 May;170(2):459-70. doi: 10.1007/s12010-013-0210-6. Epub 2013 Apr 2.
The in vitro basic biological characteristics and directed differentiation potential towards cardiomyocytes of adult adipose-derived stem cells (ADSCs) induced by angiotensin II were both investigated. ADSCs were isolated from adult adipose tissue and cultured in vitro, and were subsequently induced into adipocytes, chondrocytes, and osteoblasts for assays of multipotential differentiation. The morphological characteristics of ADSCs were observed under an inverted microscope in bright field and phase-contrast ways and a confocal laser scanning microscopy. Moreover, the directional differentiation potential was observed by Oil Red, alkaline phosphatase, von Kossa, and toluidine blue stainings, respectively. The expressions of CD34, CD44, CD45, CD105, and HLA-DR were also detected via flow cytometry. Following to this, ADSCs were induced by angiotensin II and basic fibroblast growth factor for the purpose of directional differentiation towards cardiomyocyte-like cells, and the cells treated with 5-azacytidine were regarded as the control. The results showed that the isolated and cultured ADSCs presented a typical morphology of fusiform shape and also expressed CD44, CD105, but not CD34, CD45, and HLA-DR with assays of flow cytometry. The multi-differentiations to adipocytes, chondrocytes, and osteoblasts confirmed that the isolated cells maintained the stem characteristics generating from adipose tissues. After 4 weeks of induction by angiotensin II, the cells expressed myosin heavy chain, troponin I, and connexin43 by immunocytochemistry staining, but without beating of the cells. This current study indicated that ADSCs possessed the characteristics of mesenchymal stem cells and angiotensin II could induce ADSCs into cardiomyocyte-like cells.
本研究旨在探讨血管紧张素Ⅱ（angiotensin II）诱导的成年脂肪来源干细胞（adipose-derived stem cells，ADSCs）的体外基本生物学特性和向心肌细胞的定向分化潜能。从成年脂肪组织中分离 ADCS 并进行体外培养，然后诱导其分化为脂肪细胞、软骨细胞和成骨细胞，以检测多能分化潜能。倒置显微镜下观察 ADCS 的形态特征，包括明场和相差方式，以及共聚焦激光扫描显微镜。此外，通过油红、碱性磷酸酶、茜素红和甲苯胺蓝染色分别观察定向分化潜能。流式细胞术检测 CD34、CD44、CD45、CD105 和 HLA-DR 的表达。随后，ADSCs 用血管紧张素Ⅱ和碱性成纤维细胞生长因子诱导向心肌样细胞分化，用 5-氮杂胞苷处理的细胞作为对照。结果显示，分离培养的 ADCS 呈典型的梭形形态，流式细胞术检测 CD44、CD105 表达，而 CD34、CD45 和 HLA-DR 不表达。向脂肪细胞、软骨细胞和成骨细胞的多向分化证实了分离细胞保持了源自脂肪组织的干细胞特性。用血管紧张素Ⅱ诱导 4 周后，免疫细胞化学染色显示细胞表达肌球蛋白重链、肌钙蛋白 I 和连接蛋白 43，但细胞无搏动。本研究表明 ADCS 具有间充质干细胞的特征，血管紧张素Ⅱ可诱导 ADCS 向心肌样细胞分化。
