Yoshimura Kotaro, Shigeura Tomokuni, Matsumoto Daisuke, Sato Takahiro, Takaki Yasuyuki, Aiba-Kojima Emiko, Sato Katsujiro, Inoue Keita, Nagase Takashi, Koshima Isao, Gonda Koichi
Department of Plastic Surgery, University of Tokyo School of Medicine, Bunkyo-ku, Tokyo, Japan.
J Cell Physiol. 2006 Jul;208(1):64-76. doi: 10.1002/jcp.20636.
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 (PLA) 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+ CD105low CD146+), 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.
抽脂吸出物（主要为盐溶液、血液和脂肪组织碎片）可分离为脂肪部分和液体部分。从脂肪部分分离出的细胞称为处理过的抽脂吸出物（PLA）细胞，其中包含脂肪来源的贴壁基质细胞（ASC）。在这里，我们将从抽脂吸出物液体部分分离出的细胞定义为抽脂吸出物液体（LAF）细胞。分别从这两个部分分离出基质血管成分（SVF），并在培养和未培养条件下对其进行表征。新鲜分离出的LAF细胞和PLA细胞数量相当，但贴壁的LAF细胞较少。新鲜LAF分离物中的CD34 + CD45-细胞通过贴壁培养得以扩增，这表明LAF细胞含有ASC。尽管新鲜分离的PLA细胞和LAF细胞具有不同的细胞表面标志物谱，但贴壁的PLA细胞和LAF细胞在生长动力学、形态、分化能力和表面标志物谱方面具有相当相似的特征。接种后，PLA细胞和LAF细胞均显示CD29、CD44、CD49d、CD73、CD90、CD105和CD151的表达显著增加，而CD31和CD45的表达降低。多色流式细胞术分析显示，SVF由异质细胞群体组成，包括血液来源的细胞（CD45 +）、ASC（CD31- CD34 + CD45- CD90 + CD105- CD146-）、内皮（祖）细胞（CD31 + CD34 + CD45- CD90 + CD105low CD146 +）、周细胞（CD31- CD34- CD45- CD90 + CD105- CD146 +）和其他细胞。接种后，ASC的CD105表达显著增加。尽管随着培养时间的增加，一些贴壁的ASC失去了CD34表达，但我们的培养方法使ASC中的CD34表达至少维持了10 - 20周。这些结果表明，抽脂来源的细胞可能对基于细胞的治疗有用且有价值。
