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首页> 外文期刊>Cytotherapy >An alternative method for the isolation of mesenchymal stromal cells derived from lipoaspirate samples
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An alternative method for the isolation of mesenchymal stromal cells derived from lipoaspirate samples

机译:分离源自脂肪抽吸物样品的间充质基质细胞的另一种方法

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摘要

Background aims Since initial methods were developed for isolating cells from adipose tissue, little has been done to improve mesenchymal stromal cell (MSC) yield. The aim of the present study was to isolate a population of MSC from lipoaspirate samples without tissue digestion and to assess the possibility of cryopreserving the freshly isolated cells. Methods A population of MSC was isolated from 13 patients' lipoaspirate samples by mechanical dissociation. Mechanically processed lipoaspirate adipose tissue (MPLA) cells were characterized after in vitro cell expansion by morphologic analysis, expression of MSC surface markers and differentiation assays. Results Mechanical dissociation yielded a large quantity of adherent MSC both after standard and vibro-assisted liposuction. Preservation of lipoaspirate samples at 4 C for 1 or 2 days until the mechanical procedure did not change the MPLA cell content. It was possible to store freshly isolated MPLA cells by cryopreservation without loss of the MSC population. Adherent MPLA cells were negative for CD45 and CD31 and positive for CD34, CD105, CD44 and CD90. They also showed adipogenic, osteogenic and chondrogenic potentials similar to MSC populations from other sources as already described in the literature. Conclusions MSC can be isolated from human lipoaspirate samples by the mechanical procedure described in this study with a significant reduction in time and cost. Together with cryopreservation of freshly isolated MPLA cells, this has made it easier to harvest and store MSC for therapeutic applications such as soft-tissue augmentation and tissue engineering.
机译:背景目的由于开发了用于从脂肪组织中分离细胞的初始方法,因此很少进行提高间充质基质细胞(MSC)产量的工作。本研究的目的是在不进行组织消化的情况下,从脂肪抽吸物样品中分离出MSC群体,并评估冷冻保存新鲜分离的细胞的可能性。方法采用机械离解法从13例患者的脂肪抽吸物中分离出MSC。机械加工的脂肪抽吸物脂肪组织(MPLA)细胞在体外细胞扩增后通过形态分析,MSC表面标志物的表达和分化分析进行了表征。结果机械解离在标准吸脂和振动吸脂后均产生大量粘附的MSC。将脂肪抽吸物样品在4 C下保存1或2天,直到机械操作未改变MPLA细胞含量。通过冷冻保存可以储存新鲜分离的MPLA细胞,而不会丢失MSC群体。粘附的MPLA细胞CD45和CD31阴性,而CD34,CD105,CD44和CD90阳性。他们还显示出与其他来源的MSC群体相似的成脂,成骨和成软骨潜能,正如文献中已经描述的那样。结论可以通过本研究中描述的机械方法从人抽脂样品中分离MSC,从而显着减少时间和成本。与新鲜分离的MPLA细胞的冷冻保存一起,这使得更易于收获和储存MSC,用于诸如软组织扩增和组织工程的治疗应用。

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