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The impact of cell source, culture methodology, culture location, and individual donors on gene expression profiles of bone marrow-derived and adipose-derived stromal cells

机译:细胞来源,培养方法,培养位置和单个供体对骨髓源性和脂肪源性基质细胞基因表达谱的影响

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摘要

Bone marrow (BM) stromal cells (MSCs), also known as mesenchymal stem cells, display a high degree of heterogeneity. To shed light on the causes of this heterogeneity, MSCs were collected from either human BM (n=5) or adipose tissue (AT) (n=5), and expanded using 2 different culture methods: one based on fetal calf serum, and one based on human platelet lysate. After initial expansion, MSCs were frozen, and the vials were transported to 3 different laboratories and grown for 1 passage using the same brand of culture plastic, medium, and supplements. Subsequently, the cells were harvested and assayed for their gene expression profile using the Affymetrix exon microarray platform. Based on gene expression profiles, the most discriminative feature was the anatomical harvesting site, followed by culture methodology. Remarkably, genes in the WNT pathway were expressed at higher levels in BM-derived MSCs than in AT-derived MSCs. Although differences were found between laboratories, cell culture location only slightly affects heterogeneity. Furthermore, individual donors contributed marginally to the observed differences in transcriptomes. Finally, BM-derived MSCs displayed the highest level of similarity, irrespective their culture conditions, when compared to AT-derived cells.
机译:骨髓(BM)基质细胞(MSC),也称为间充质干细胞,表现出高度的异质性。为了阐明这种异质性的原因,从人BM(n = 5)或脂肪组织(AT)(n = 5)收集MSC,并使用两种不同的培养方法进行扩增:一种基于胎牛血清,一种基于人血小板裂解物。最初扩增后,将MSC冷冻,然后将小瓶运输到3个不同的实验室,并使用相同品牌的培养塑料,培养基和补充剂培养1代。随后,收获细胞,并使用Affymetrix外显子微阵列平台分析其基因表达谱。根据基因表达谱,最有区别的特征是解剖收获部位,其次是培养方法。值得注意的是,WNT途径中的基因在BM来源的MSC中的表达水平高于AT来源的MSC。尽管在实验室之间发现了差异,但细胞培养位置仅轻微影响异质性。此外,单个供体仅对观察到的转录组差异有所贡献。最后,与AT来源的细胞相比,无论其培养条件如何,BM来源的MSC都显示出最高的相似性。

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