Urbani S, Caporale R, Lombardini L, Bosi A, Saccardi R
Hematology Unit, Careggi Hospital, Viale Morgagni 85, 50134 Florence, Italy.
Cytotherapy. 2006;8(3):243-53. doi: 10.1080/14653240600735834.
Mesenchymal stem cells (MSC) are multipotent progenitors retaining the capability to undergo multilineage differentiation, mostly towards all the mesodermal cellular lineages. MSC growing under standard conditions are composed of two main subpopulations with a characteristic distribution in the morphologic flow cytometric scatter: RS (recycling stem) cells (small, agranular) and m (mature) MSC (large, moderately granular cells).
MSC obtained from BM of healthy donors and expanded in culture were characterized by evaluating both the expression of conventional markers and differentiation potential. We used CFSE, a lipophilic dye that is taken up by cell membranes, to investigate separately the proliferative activity of RS cells and mMSC subsets.
With flow cytometric analysis, RS cells and mMSC showed nearly the same immunophenotypic pattern, even if a significantly smaller percentage of RS cells expressed some of the classic mesenchymal Ag. The RS cell fraction was confirmed to have a higher proliferative potential and such a feature was particularly evident under certain culture conditions.
CFSE has been shown as a reliable method for studying the proliferative activity of MSC subpopulations identified by flow cytometric analysis. The acquisition parameter strategy is crucial for the accuracy of the analysis.
间充质干细胞（MSC）是多能祖细胞，保留了进行多谱系分化的能力，主要分化为所有中胚层细胞谱系。在标准条件下生长的MSC由两个主要亚群组成，在形态学流式细胞术散射图中有特征性分布：RS（循环干细胞）细胞（小，无颗粒）和m（成熟）MSC（大，中度颗粒细胞）。
从健康供体的骨髓中获取并在培养中扩增的MSC，通过评估传统标志物的表达和分化潜能进行表征。我们使用CFSE，一种可被细胞膜摄取的亲脂性染料，来分别研究RS细胞和mMSC亚群的增殖活性。
通过流式细胞术分析，RS细胞和mMSC显示出几乎相同的免疫表型模式，即使RS细胞中表达某些经典间充质抗原的比例明显较小。RS细胞部分被证实具有更高的增殖潜能，并且在某些培养条件下这种特征尤为明显。
CFSE已被证明是研究通过流式细胞术分析鉴定的MSC亚群增殖活性的可靠方法。采集参数策略对分析的准确性至关重要。
