Sobh Mohamed A
Department of Zoology, Urology and Nephrology Center, Research Center, Mansoura University, Cairo, Egypt.
Anat Cell Biol. 2014 Jun;47(2):83-90. doi: 10.5115/acb.2014.47.2.83. Epub 2014 Jun 20.
Mesenchymal stem cells (MSCs) offer significant promise as a multipotent source for cell-based therapies and could form the basis for the differentiation and cultivation of tissue grafts to replace damaged tissue. However, no gene expression follow up analysis has been undertaken to characterize the in vitro adipogenic differentiated MSCs. The main goal of this study was to focus on MSCs and to analyze their differentiation capacity. To achieve this aim, bone marrow MSCs from sprague dawely rats were isolated, expanded in monolayer culture and characterized with respect to their cluster of differentiation (CD) and ability for adipogenic differentiation capacity. The expression of CD44, CD45, CD29, CD34, and CD90 on bone marrow derived MSCs was characterized using flow cytometry. Adipogenesis was determined by staining with oil-red O and reverse transcription polymerase chain reaction assessments of lipoprotein lipase, leptin, adiponectin and adipocyte genes at different time intervals, after 4, 7, 14, and 21 days. Our results revealed that the pattern of CD marker expression was highly positive significant with CD29, CD44, and CD90 when compared with CD34 and CD45. MSCs showed proliferative potential and were capable of adipogenic differentiation characterized by reddish brown-droplets following staining with oil-red O and expression of molecular bands of genes. These results demonstrate, at the morphological, immunophenotyping and gene expression levels, the multipotency of MSCs and thus highlight their potential therapeutic value for cell-based tissue engineering.
间充质干细胞(MSCs)作为基于细胞的治疗的多能来源具有巨大潜力,可为组织移植的分化和培养奠定基础,以替代受损组织。然而,尚未进行基因表达后续分析来表征体外脂肪生成分化的间充质干细胞。本研究的主要目标是聚焦于间充质干细胞并分析其分化能力。为实现这一目标,从Sprague Dawely大鼠中分离出骨髓间充质干细胞,在单层培养中进行扩增,并就其分化簇(CD)和脂肪生成分化能力进行表征。使用流式细胞术对骨髓来源的间充质干细胞上CD44、CD45、CD29、CD34和CD90的表达进行表征。在4、7、14和21天的不同时间间隔后,通过油红O染色以及脂蛋白脂肪酶、瘦素、脂联素和脂肪细胞基因的逆转录聚合酶链反应评估来确定脂肪生成。我们的结果显示,与CD34和CD45相比,CD标志物表达模式在CD29、CD44和CD90方面呈高度阳性显著。间充质干细胞显示出增殖潜力,并能够进行脂肪生成分化,其特征在于用油红O染色后出现红棕色液滴以及基因分子条带的表达。这些结果在形态学、免疫表型和基因表达水平上证明了间充质干细胞的多能性,从而突出了它们在基于细胞的组织工程中的潜在治疗价值。