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人脂肪来源干细胞增殖和肝向分化的比较研究。

A comparative study of proliferation and hepatic differentiation of human adipose-derived stem cells.

机构信息

Nanobiosciences Unit, JRC - European Commission Institute for Health and Consumer Protection, Ispra, Italy.

出版信息

Cells Tissues Organs. 2010;191(6):466-77. doi: 10.1159/000273266. Epub 2009 Dec 30.

Abstract

Human adipose-derived stem cells possess a lot of stem cell characteristics, so they may be considered a source of stem cell population. On the basis of that, we have investigated the hepatic potential of adipose-derived stem cells, obtained from liposuction, following two differentiation protocols. In the first procedure, medium was supplemented with epidermal growth factor (EGF), basic fibroblast growth factor, hepatocyte growth factor (HGF) and nicotinamide; the second involved the addition of factors such as dexametasone, EGF, insulin-transferrin-sodium selenite, HGF, dimethyl sulfoxide and oncostatin. In parallel, we carried out our study in the Hep G2 cell line, as human hepatic differentiated in vitro model. Immunocytochemical analysis and RT-PCR were performed using hepatic markers to evaluate cell differentiation. DNA content, MTT test and carboxyl fluorescein succinimidyl ester staining were carried out to evaluate cell proliferation. We reported the evidence of basal hepatic marker in undifferentiated adipose-derived stem cells, which confirmed their multipotency. A strong expression of albumin and alpha-fetoprotein was observed in hepatic-induced adipose-derived stem cells following both differentiation procedures. Morphological aspects of the two types of hepatic adipose-derived stem cells were alike. Proliferation index suggested that the first differentiation procedure promoted better growth than the second. These preliminary findings suggest adipose-derived stem cells may be induced into hepatic lineage, and the most significant difference between the two standard differentiation procedures concerns proliferation rate. This aspect is to be considered when adipose-derived stem cells are employed in research and clinical studies.

摘要

人脂肪来源的干细胞具有许多干细胞的特征,因此可以被视为干细胞群体的来源。在此基础上,我们通过两种分化方案研究了从吸脂中获得的脂肪来源干细胞的肝潜能。在第一种方案中,培养基中添加了表皮生长因子(EGF)、碱性成纤维细胞生长因子、肝细胞生长因子(HGF)和烟酰胺;第二种方案涉及添加地塞米松、EGF、胰岛素-转铁蛋白-亚硒酸钠、HGF、二甲基亚砜和肿瘤坏死因子等因子。同时,我们在 Hep G2 细胞系中进行了我们的研究,作为体外分化的人类肝模型。使用肝标志物进行免疫细胞化学分析和 RT-PCR,以评估细胞分化。进行 DNA 含量、MTT 试验和羧基荧光素琥珀酰亚胺酯染色,以评估细胞增殖。我们报告了未分化的脂肪来源干细胞中存在基础肝标志物的证据,这证实了它们的多能性。在两种分化程序后,诱导的脂肪来源干细胞中观察到白蛋白和甲胎蛋白的强烈表达。两种类型的肝诱导脂肪来源干细胞的形态学方面相似。增殖指数表明,第一种分化方案比第二种方案更能促进生长。这些初步发现表明脂肪来源的干细胞可以被诱导为肝谱系,两种标准分化程序之间的最显著差异涉及增殖率。在将脂肪来源的干细胞用于研究和临床研究时,应考虑这一方面。

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