"In vitro" and multicolor phenotypic characterization of cell subpopulations identified in fresh human adipose tissue stromal vascular fraction and in the derived mesenchymal stem cells
"In vitro" and multicolor phenotypic characterization of cell subpopulations identified in fresh human adipose tissue stromal vascular fraction and in the derived mesenchymal stem cells
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DOI：
10.1186/1479-5876-5-55
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发表时间：
2007-10-31
影响因子：
7.4
通讯作者：
Moccetti, Tiziano
中科院分区：
文献类型：
--
作者：
Astori, Giuseppe;Vignati, Francesca;Moccetti, Tiziano
背景：间质血管成分（stromal vascular fraction，SVF）是来源于脂肪组织的异质性细胞群.仍然缺乏关于构成SVF的细胞亚群的表征以及其间充质和造血潜能的信息。方法：采用流式细胞仪分析、细胞学和体外实验等方法对SVF的组成进行研究。我们用流式细胞仪（FACS）和人CFC（colony-forming-cell haematopoietic assay）相结合的方法研究了CD 34+细胞的数量。通过定量特异性标志物（CD 146、CD 105、CD 31和UEA-I）的共表达来研究内皮分数。通过CFU-F测定评估间充质潜能，并通过5色FACS分析表征培养的AT-MSC。多能分化潜能（成骨，脂肪和软骨形成）进行了调查，在细胞和分子水平。结果：我们确定了在SVF两个CD 34+群体的抗原表达强度有显着差异，大多数细胞表达CD 34在低强度。此外，两个CD 146+细胞群体在SVF中是明显可区分的：占总SVF细胞的9.9%的CD 146 dim和占约39.3%的CD 146 + bright细胞群体。CFC克隆的频率与外周血报告的频率相当。内皮细胞约占SVF细胞的7.7%。结论：SVF不是一个均一的细胞群，其最终组成可能受到流式细胞术分析和SVF提取步骤的影响。SVF中的CFU-F频率为1/4880，该值约为骨髓报告数据的7倍。AT-MSC的抗原谱与骨髓来源的MSC相当。AT-MSC能够沿着成骨、成脂肪和成软骨谱系分化。本文报道的数据进一步有助于SVF的表征，SVF是一种为临床应用提供替代MSC来源的组织。
Background: The stromal vascular fraction (SVF) is a heterogeneous cell population derived from the adipose tissue. There is still a lack of information concerning the characterization of the cell subpopulations constituting the SVF as well as its mesenchymal and haematopoietic potential. Furthermore there are great variations in its phenotypical characterization.Methods: Composition of SVF was investigated by FACS analysis, cytological and "in vitro" assays. We studied CD34+ population by combining FACS with human CFC (colony-forming-cell haematopoietic assay). The endothelial fraction was investigated by quantifying the co-expression of specific markers (CD146, CD105, CD31 and UEA-I). Mesenchymal potential was assessed by CFU-F assay and cultured AT-MSC were characterized by a 5-color FACS analysis. The multipotent differentiation potential (osteogenic, adipogenic and chondrogenic) was investigated both at cellular and molecular level.Results: We identified in the SVF two CD34+ populations with a marked difference in the intensity of antigen expression, the majority of the cells expressing CD34 at low intensity. Moreover, two CD146+ cell populations were clearly distinguishable in the SVF: a CD146 dim accounting for 9.9% of the total SVF cells and a CD146+ bright cell population accounting for about 39.3%. The frequency of CFC clones was comparable with the one reported for peripheral blood. Endothelial cells account for about 7.7% of the SVF cells. AT-MSC differenced in the osteogenic adipogenic and chondrogenic lineage.Conclusion: The SVF is not a homogeneous cell population, and its final composition could be influenced both by the flow cytometric technique analysis and the SVF extraction steps. The CFU-F frequency in the SVF was 1/4880, a value about seven times greater than the data reported for bone marrow. The antigenic profile of AT-MSC was comparable with bone-marrow derived MSC. AT-MSC were able to differentiate along the osteogenic adipogenic and chondrogenic lineages. The data here reported, further contribute to the characterization of SVF, a tissue providing an alternative as a source of MSC for clinical applications.