Isolation of BM mesenchymal stem cells by plastic adhesion or negative selection: phenotype, proliferation kinetics and differentiation potential
Isolation of BM mesenchymal stem cells by plastic adhesion or negative selection: phenotype, proliferation kinetics and differentiation potential
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DOI：
10.1080/14653240410004943
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发表时间：
2004-08-01
期刊：
影响因子：
4.5
通讯作者：
Bron, D
中科院分区：
文献类型：
--
作者：
Tondreau, T;Lagneaux, L;Bron, D
背景骨髓间充质干细胞（mesenchymal stem cells，MSC）具有更新和分化为多种组织的潜能。在这项研究中，我们比较了不同的富集方法，以获得骨髓间充质干细胞从BM。MethodsThree不同的方法进行了比较，以期更迅速地获得骨髓间充质干细胞：阴性选择（RosetteSep和MACS）和塑料粘附。三种细胞组分在完全α-最低必需培养基中生长，以评价其增殖能力、培养期间的表型及其分化成脂肪细胞、骨细胞和软骨细胞的潜力。MSC的鉴定进行了免疫荧光与假定的间充质标记SH 2和SH 3，但也与造血marker.ResultsAfter阴性选择，只有1 +/- 0.2%和2.9 +/- 0.8%的细胞从BM回收与RosetteSep和MACS方法，分别。然而，负去除允许在培养10天后快速且大量地获得具有超过90%的SH 2（+）和SH 3（+）细胞的MSC的同质群体。如果使用塑料粘附作为选择方法，则在三次传代后以及在平均25 - 30天的培养后观察到类似的均一性。Stro-1表达水平的变化也表明了MSC成熟程度的不同。讨论RosetteSep耗竭选择法可能是一种从骨髓中快速获得MSC的简单方法，其目的是潜在的治疗用途。
BackgroundBM mesenchymal stem cells ( MSC) have the capacity for renewal and the potential to differentiate into multiple tissues. In this study, we compared different enrichment methods to obtain MSC from BM.MethodsThree different methods were compared with a view to obtaining MSC more rapidly from BM: negative selection (RosetteSep and MACS) and plastic adhesion. The three cell fractions were grown in complete alpha-minimum essential medium in order to evaluate their proliferative capacity, their phenotype during culture and their potential to differentiate into adipocytes, osteocytes and chondrocytes. Identification of MSC was performed by immunofluorescence with putative mesenchymal markers SH2 and SH3 but also with hematopoietic markers.ResultsAfter negative selection, only 1 +/- 0.2% and 2.9 +/- 0.8% of cells were recovered from BM with the RosetteSep and MACS methods, respectively. However, negative depletion permitted a homogeneous population of MSC, with more than 90% SH2(+) and SH3(+) cells, to be obtained rapidly and in large quantities after 10 days of culture. Similar homogeneity was observed after three passages if the plastic adhesion was used as selection method and after an average of 25 - 30 days of culture. Different levels of MSC maturity were also suggested by the variable level expression of Stro-1.DiscussionDepleting selection by RosetteSep may represent an easy method of obtaining MSC rapidly from BM with the aim of potential therapeutic use.