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.

吸脂剂吸出（主要是盐水溶液，血液和脂肪组织碎片）分为脂肪和液体部分。从脂肪部分分离出的细胞称为脂式脂肪酸（PTA）细胞，并包含脂肪衍生的粘附基质细胞（ASC）。在这里，我们定义从吸脂液的流体部分分离为吸脂液（LAF）细胞的细胞。基质血管级分（SVF）分别与两部分分开，并在培养和非培养条件下进行表征。新鲜分离的LAF和PLA细胞数量可比数量，但较少的LAF细胞是粘附的。来自新鲜LAF分离株的CD34+CD45细胞通过粘附培养物扩展，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，但在生长动力学，形态，分化能力和表面标记物谱方面，粘附的PLA和LAF细胞具有非常相似的特征。电镀后，PLA和LAF细胞均显示CD29，CD44，CD49D，CD73，CD90，CD105和CD151的表达显着增加，并且CD31和CD45的表达降低。多色FACS分析表明，SVF由异构细胞群组成，包括血液衍生细胞（CD45+），ASC（CD31-CD34+CD45-CD90+CD105-CD146-），内皮（祖细胞）（CD31+CD34+CD34+CD45-CD90+CD45-CD90+CD45-CD90+CD114660146）（CD31-CD34-CD45-CD90+CD105-CD146+）和其他单元。电镀后，ASC在CD105表达中显示出急剧增加。尽管一些依从性ASC随着培养时间的增加而失去了CD34的表达，但我们的培养方法至少在ASC中保持CD34的表达至少10-20周。这些结果表明，吸脂细胞可能对基于细胞的疗法有用且有价值。