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Isolation and transcription profiling of purified uncultured human stromal stem cells: Alteration of gene expression after in vitro cell culture

机译:纯化的未培养人基质干细胞的分离和转录谱:体外细胞培养后基因表达的改变

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摘要

Stromal stem cells proliferate in vitro and may be differentiated along several lineages. Freshly isolated, these cells have been too few or insufficiently pure to be thoroughly characterized. Here, we have isolated two populations of CD45(-)CD34(+)CD105(+) cells from human adipose tissue which could be separated based on expression of CD31. Compared with CD31(+) cells, CD31(-) cells overexpressed transcripts associated with cell cycle quiescence and sternness, and transcripts involved in the biology of cartilage, bone, fat, muscle, and neural tissues. In contrast, CD31(+) cells overexpressed transcripts associated with endothelium and the major histocompatibility complex class II complex. Clones of CD31(-) cells could be expanded in vitro and differentiated into cells with characteristics of bone, fat, and neural-like tissue. On culture, transcripts associated with cell cycle quiescence, sternness, certain cytokines and organ specific genes were down-regulated, whereas transcripts associated with signal transduction, cell adhesion, and cytoskeletal components were up-regulated. CD31(+) cells did not proliferate in vitro. CD45(-)CD34(+)CD105(+)CD31(-) cells from human adipose tissue have stromal stem cell properties which may make them useful for tissue engineering.
机译:基质干细胞在体外增殖,并可沿多个谱系分化。这些细胞是新鲜分离的,纯度太低或纯度不足,无法进行全面鉴定。在这里,我们从人类脂肪组织中分离了两个种群的CD45(-)CD34(+)CD105(+)细胞,这些细胞可以根据CD31的表达进行分离。与CD31(+)细胞相比,CD31(-)细胞过度表达与细胞周期静止和严峻有关的转录本,以及与软骨,骨骼,脂肪,肌肉和神经组织生物学有关的转录本。相反,CD31(+)细胞过度表达与内皮和主要组织相容性复合物II类复合物相关的转录物。 CD31(-)细胞的克隆可以在体外扩增,并分化为具有骨骼,脂肪和神经样组织特征的细胞。在培养中,与细胞周期静止,严峻性,某些细胞因子和器官特异性基因相关的转录本被下调,而与信号转导,细胞粘附和细胞骨架成分相关的转录本被上调。 CD31(+)细胞不能在体外增殖。来自人脂肪组织的CD45(-)CD34(+)CD105(+)CD31(-)细胞具有基质干细胞特性,这可能使它们可用于组织工程。

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