Study on tissue and vascular regeneration using adipose-derived stem cells.

利用脂肪干细胞进行组织和血管再生的研究。

基本信息

批准号：
16390507
负责人：
YOSHIMURA Kotaro
金额：
$ 8.9万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

项目摘要

Liposuction aspirates (primarily saline solution, blood, and adipose tissue fragments) separate into fatty and fluid portions. Cells isolated from the fatty portion are termed processed lipoaspirate (PTA) cells and contain adipose-derived adherent stromal cells (ASCs). Here we define cells isolated from the fluid portion of liposuction aspirates as liposuction aspirate fluid (LAF) cells. Stromal vascular fractions (SVF) were isolated separately from both portions and characterized under cultured and non-cultured conditions. A comparable number of LAF and PLA cells were freshly isolated, but fewer LAF cells were adherent. CD34+CD45- cells from fresh LAF isolates were expanded by adherent culture, suggesting that LAF cells contain ASCs. Although freshly isolated PLA and LAF cells have distinct cell surface marker profiles, adherent PLA and LAF cells have quite similar characteristics with regard to growth kinetics, morphology, capacity for differentiation, and surface marker profiles. After plating, both PLA and LAF cells showed significant increased expression of CD29, CD44, CD49d, CD73, CD90, CD105,and CD151 and decreased expression of CD31 and CD45. Multicolor FACS analysis revealed that SVF are composed of heterogeneous cell populations including blood-derived cells (CD45+), ASCs (CD31-CD34+CD45-CD90+CD105-CD146-), endothelial (progenitor) cells (CD31+CD34+CD45-CD90+CD105lowCD146+), pericytes (CD31-CD34-CD45-CD90+CD105-CD146+), and other cells. After plating, ASCs showed a dramatic increase in CD105 expression. Although some adherent ASCs lost CD34 expression with increasing culture time, our culture method maintained CD34 expression in ASCs for at least 10-20 weeks. These results suggest that liposuction-derived cells may be useful and valuable for cell-based therapies.

抽脂抽吸物（主要是盐水溶液、血液和脂肪组织碎片）分离成脂肪和液体部分。从脂肪部分分离的细胞被称为加工后的吸脂 (PTA) 细胞，并含有脂肪来源的贴壁基质细胞 (ASC)。在这里，我们将从抽脂抽吸液的液体部分分离的细胞定义为抽脂抽吸液（LAF）细胞。基质血管组分（SVF）分别从两个部分中分离出来，并在培养和非培养条件下进行表征。新鲜分离的 LAF 和 PLA 细胞数量相当，但贴壁的 LAF 细胞较少。来自新鲜 LAF 分离株的 CD34+CD45- 细胞通过贴壁培养进行扩增，表明 LAF 细胞含有 ASC。尽管新鲜分离的 PLA 和 LAF 细胞具有不同的细胞表面标记谱，但贴壁的 PLA 和 LAF 细胞在生长动力学、形态、分化能力和表面标记谱方面具有非常相似的特征。铺板后，PLA 和 LAF 细胞均表现出 CD29、CD44、CD49d、CD73、CD90、CD105 和 CD151 表达显着增加，而 CD31 和 CD45 表达显着降低。多色 FACS 分析显示 SVF 由异质细胞群组成，包括血源细胞 (CD45+)、ASC (CD31-CD34+CD45-CD90+CD105-CD146-)、内皮（祖）细胞 (CD31+CD34+CD45-CD90) +CD105lowCD146+), 周细胞(CD31-CD34-CD45-CD90+CD105-CD146+)和其他细胞。铺板后，ASC 显示 CD105 表达显着增加。尽管一些贴壁ASC随着培养时间的增加而失去CD34表达，但我们的培养方法在ASC中维持CD34表达至少10-20周。这些结果表明，吸脂衍生的细胞对于基于细胞的疗法可能是有用且有价值的。