旧把戏的新把戏:脂肪组织中人间充质干细胞的顺磁细胞分选

J. Murray, A. Doty, Noah J. Jones, Marie Crandall, E. Scott
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引用次数: 2

摘要

脂肪组织来源的间充质干细胞(ASCs)可以在没有体外扩增的情况下分离出临床有用的数量。本研究的目的是验证一种利用顺磁珠富集原生ASCs的新方法。兔抗小鼠原代抗体与顺磁微珠结合。二抗,选择性ASCs,然后结合到一抗构建所谓的顺磁免疫珠(PIBs)。然后将pib添加到新鲜的人抽脂液中,在10分钟内形成ASC-PIB偶联物(aPIBS)。然后将手持式磁铁放置在抽脂液- apib混合物附近,在接下来的10分钟内,沉淀apib。每mL抽脂液活细胞计数为9.6 × 104。扫描电镜显示与api相一致的沉淀物。流式细胞术鉴定出CD90和CD105的细胞结合标记,而培养证实了三系分化,所有这些都是诊断ASCs的属性。本研究证实磁富集法可在20分钟内从抽脂液中分离出功能性ASCs。由于通过吸脂和这种ASC富集技术获取脂肪组织都不需要电,因此新鲜的初级治疗性ASC可以在任何护理点环境中分离出来,甚至在发展中国家也是如此。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A New Ball for an Old Trick: Paramagnetic Cell Sorting of Human Mesenchymal Stem Cells from Adipose Tissue
Adipose tissue-derived mesenchymal stem cells (ASCs) may be isolated in clinically useful quantities without in vitro expansion. The purpose of this study is to validate a novel method for the enrichment of primary ASCs using paramagnetic beads. Primary rabbit anti-mouse antibodies were bound to paramagnetic microbeads. Secondary antibodies, selective for ASCs, were then bound to the primary antibodies to construct so-called paramagnetic immunobeads (PIBs). PIBs were then added to fresh human lipoaspirate to create ASC-PIB conjugates (aPIBS) over 10 minutes. A hand-held magnet was then placed adjacent to the lipoaspirate-aPIBs mixture, and over the next 10 minutes, the aPIBs were precipitated. Live cell count per mL of lipoaspirate was 9.6 x 104. Scanning electron microscopy revealed precipitates consistent with aPIBs. Flow cytometry identified cell-bound markers for CD90 and CD105 while culture confirmed tri-lineage differentiation, all attributes diagnostic of ASCs. This study validates that functional ASCs may be isolated from lipoaspirate by magnetic enrichment in 20 minutes. As both the harvest of adipose tissue by liposuction and this ASC enrichment technique do not require electricity, fresh primary therapeutic ASCs may be isolated in any point-of-care setting, even in developing countries.
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