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Explant culture: an efficient method to isolate adipose-derived stromal cells for tissue engineering.

机译:外植体培养:一种分离脂肪来源的基质细胞用于组织工程的有效方法。

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摘要

Enzymatic digestion, the commonly used method of adipose-derived stromal cells isolation, is time consuming and expensive, especially when applied to large volumes of tissue. In the present study, the characteristics of the cells obtained by adipose tissue explant culture were studied. We found that adipose tissue fragments could adhere onto the growth surface of flasks in a very short time after plating and that fibroblast-like cells migrated from the explants and reached confluence. Morphologic analysis and surface markers expression suggested the mesenchymal origin of the cells derived from adipose tissue explants. After in vitro expansion these cells were successfully induced into adipogenic, osteogenic, and chondrogenic lineages, which demonstrated their multipotency. The high growth rate and colony-forming efficiency of explant-derived cells were similar to those of cells obtained by digestion. Furthermore, explant culture gave higher yield of cells than digestion method after primary culture. The experiment of ectopic adipogenesis in nude mice suggested the prospects for tissue engineering of these cells. In conclusion, we obtained multipotent stromal cells from adipose tissue by explant culture, and this method was simple, time saving, and gave a high yield of cells. Therefore, explant culture can be used as an effective way to isolate adipose-derived stromal cells for tissue engineering.
机译:酶促消化是脂肪来源的基质细胞分离的常用方法,既耗时又昂贵,特别是应用于大量组织时。在本研究中,研究了通过脂肪组织外植体培养获得的细胞的特性。我们发现,在铺板后很短的时间内,脂肪组织碎片可能会粘附在烧瓶的生长表面上,并且成纤维细胞样细胞从外植体迁移并达到汇合。形态学分析和表面标志物表达提示源自脂肪组织外植体的细胞的间充质起源。在体外扩增后,这些细胞被成功诱导为成脂,成骨和成软骨谱系,证明了它们的多能性。外植体来源的细胞的高生长速率和集落形成效率与通过消化获得的细胞相似。此外,外植体培养比初次培养后的消化方法具有更高的细胞产率。裸鼠异位脂肪形成的实验提示了这些细胞的组织工程学前景。总之,我们通过外植体培养从脂肪组织中获得了多能干基质细胞,该方法简便,省时,并且细胞产量高。因此,外植体培养可用作分离脂肪来源的基质细胞用于组织工程的有效方法。

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