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

吸脂剂（主要是生理盐水、血液和脂肪组织碎片）分为脂肪部分和液体部分。从脂肪部分分离的细胞称为加工抽脂细胞（PTA），含有脂肪来源的贴壁基质细胞（ASCs）。在这里，我们将从吸脂器液体部分分离的细胞定义为吸脂液（LAF）细胞。基质血管组分（SVF）分别从两个部分分离，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。新鲜LAF分离物的CD34+CD45-细胞通过贴壁培养扩增，提示LAF细胞含有ASCs。虽然新鲜分离的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。镀后，PLA和LAF细胞CD29、CD44、CD49d、CD73、CD90、CD105和CD151的表达均显著升高，CD31和CD45的表达均显著降低。多色FACS分析显示，SVF由异质细胞群组成，包括血源性细胞（CD45+）、ASCs （CD31-CD34+CD45- cd90 +CD105-CD146-）、内皮细胞（祖细胞）（CD31+CD34+CD45- cd90 +CD105lowCD146+）、周细胞（CD31-CD34-CD45- cd90 +CD105-CD146+）和其他细胞。移植后，ASCs的CD105表达显著增加。尽管随着培养时间的增加，一些粘附的ASCs失去了CD34的表达，但我们的培养方法至少在10-20周内保持了ASCs中CD34的表达。这些结果表明，吸脂源性细胞可能对基于细胞的治疗有用且有价值。

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.