Engineering Adipose Tissue from Uncultured Human Adipose Stromal Vascular Fraction on Collagen Matrix and Gelatin Sponge Scaffolds
Engineering Adipose Tissue from Uncultured Human Adipose Stromal Vascular Fraction on Collagen Matrix and Gelatin Sponge Scaffolds
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DOI：
10.1089/ten.tea.2010.0688
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发表时间：
2011-06-01
影响因子：
4.1
通讯作者：
Lai, Chung-Sheng
中科院分区：
文献类型：
--
作者：
Lin, Sin-Daw;Huang, Shu-Hung;Lai, Chung-Sheng
将未培养的基质血管部分（SVF）加载到多孔3D胶原基质和预先确定形状的明胶海绵支架上，评估其成脂潜能。从16个新鲜去脂的脂肪中分离出SVF。平均细胞数为6.0 +/- 4.68 × 10(7)个/mL，平均细胞存活率为72%。流式细胞术分析显示，SVF中存在脂肪来源的基质细胞（CD31（-）、CD34（-/+）、CD45（-）、CD90（+）、CD105（-）、CD146（-）和CD166(+)）。收获脂肪3小时后，将200亩L分离的SVF装入实验支架（重量为4mg），在Dulbecco改良Eagle培养基中培养。倒置光学显微镜和扫描电子显微镜下的检查表明，SVF细胞在孔表面有充分的播种和活跃的增殖。细胞生长成不同大小的簇状或束状。第28天，H&E染色对构建体进行组织学研究，微观结构显示有活脂肪细胞。油红O染色阳性证实成熟脂肪细胞脂质积累。通过GAPDH基因检测的基因组DNA的存在，以及过氧化物酶体增殖物激活受体- γ和脂蛋白脂肪酶的mRNA表达，进一步评估人脂肪细胞的存在。结果表明，将新鲜分离的未培养的SVF植入三维多孔胶原基质和明胶海绵支架上，可以再生新的脂肪组织。
Adipogenic potential was evaluated in uncultured stromal vascular fraction (SVF) loaded onto porous 3D collagen matrix and gelatin sponge scaffolds with predefined shapes. The SVF was isolated from 16 freshly lipectomized fat. Mean cell number was 6.0 +/- 4.68 x 10(7) cells/mL, and mean cell viability was 72%. Flow cytometric analysis revealed adipose-derived stromal cells (CD31(-), CD34(-/+), CD45(-), CD90(+), CD105(-), CD146(-), and CD166(+)) in the SVF. Three hours after harvest of fat, 200 mu L of isolated SVF was loaded onto an experimental scaffold (4mg in weight) and cultured in Dulbecco's modified Eagle's medium. Examination of the construct under an inverted light microscope and a scanning electron microscope demonstrated adequate seeding and active proliferation of the SVF cells on pore surfaces. Cells grew to varying sizes in clusters or in strands. On day 28, histologic study of the constructs by H&E staining revealed viable adipocytes in the microstructure. Positive Oil-Red O stain confirmed lipid accumulation in mature adipocytes. The presence of human adipocytes was further assessed by the presence of genomic DNA detected by GAPDH gene and by the mRNA expression of peroxisome proliferator activated receptor-gamma, and lipoprotein lipase. The results demonstrated that new adipose tissue can be regenerated by seeding freshly isolated, uncultured SVF on 3D porous collagen matrix and gelatin sponge scaffolds.