Immunophenotype of human adipose-derived cells: Temporal changes in stromal-associated and stem cell-associated markers
Immunophenotype of human adipose-derived cells: Temporal changes in stromal-associated and stem cell-associated markers
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DOI：
10.1634/stemcells.2005-0234
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发表时间：
2006-02-01
期刊：
影响因子：
5.2
通讯作者：
Gimble, Jeffrey M.
中科院分区：
文献类型：
--
作者：
Mitchell, James B.;McIntosh, Kevin;Gimble, Jeffrey M.
脂肪组织代表了多能成体干细胞的丰富和可获得的来源，并被许多研究人员用于组织工程应用;然而，并非所有实验室都在分离和传代的同等阶段使用细胞。我们比较了新鲜分离的人脂肪组织来源的基质血管成分（SVF）细胞相对于连续传代的脂肪源性干细胞（ASC）的免疫表型。初始SVF细胞以1：32的频率含有集落形成单位成纤维细胞。集落形成单位脂肪细胞和成骨细胞以相当的频率（分别为1：28和1：16）存在于SVF细胞中。流式细胞仪检测脂肪细胞的免疫表型随贴壁和传代的进行性变化。基质细胞相关标志物（CD 13、CD 29、CD 44、CD 63、CD 73、CD 90、CD 166）最初在SVF细胞上较低，并随着连续传代而显著增加。干细胞相关标志物CD 34在SVF细胞和/或早期传代ASC中处于峰值水平，并且在整个培养期间保持存在，尽管水平降低。醛脱氢酶和多药耐药转运蛋白（ABCG 2），这两者都已被用来识别和表征造血干细胞，表达的SVF细胞和ASC在可检测的水平。内皮细胞相关标志物（CD 31、CD 144或VE-钙粘蛋白、血管内皮生长因子受体2、血管性血友病因子）在SVF细胞上表达，并且在连续传代中没有显著变化。因此，与粗SVF的异质性相比，人脂肪来源的细胞在补充胎牛血清的培养基中粘附到塑料上并随后扩增选择相对均一的细胞群，富集表达基质免疫表型的细胞。
Adipose tissue represents an abundant and accessible source of multipotent adult stem cells and is used by many investigators for tissue engineering applications; however, not all laboratories use cells at equivalent stages of isolation and passage. We have compared the immunophenotype of freshly isolated human adipose tissue-derived stromal vascular fraction (SVF) cells relative to serial-passaged adipose-derived stem cells (ASCs). The initial SVF cells contained colony-forming unit fibroblasts at a frequency of 1:32. Colony-forming unit adipocytes and osteoblasts were present in the SVF cells at comparable frequencies (1:28 and 1:16, respectively). The immunophenotype of the adipose-derived cells based on flow cytometry changed progressively with adherence and passage. Stromal cell-associated markers (CD13, CD29, CD44, CD63, CD73, CD90, CD166) were initially low on SVF cells and increased significantly with successive passages. The stem cell-associated marker CD34 was at peak levels in the SVF cells and/or early-passage ASCs and remained present, although at reduced levels, throughout the culture period. Aldehyde dehydrogenase and the multidrug-resistance transport protein (ABCG2), both of which have been used to identify and characterize hematopoietic stem cells, are expressed by SVF cells and ASCs at detectable levels. Endothelial cell-associated markers (CD31, CD144 or VE-cadherin, vascular endothelial growth factor receptor 2, von Willebrand factor) were expressed on SVF cells and did not change significantly with serial passage. Thus, the adherence to plastic and subsequent expansion of human adipose-derived cells in fetal bovine serum-supplemented medium selects for a relatively homogeneous cell population, enriching for cells expressing a stromal immunophenotype, compared with the heterogeneity of the crude SVF.