Zhang Yun-song, Gao Jian-hua, Lu Feng, Zhu Ming
Department of Plastic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Zhonghua Yi Xue Za Zhi. 2008 Oct 21;88(38):2705-9.
To explore the possibility of building tissue-engineered adipose tissue and find a new approach for repairing soft tissue defects.
Using enzymatic digestion, adipose tissue-derived stem cells (ASCs) were extracted from the lipid part of human liposuction aspirate, cultured, and underwent adipogenic induction or not. The adipogenic-induced and non-adipogenic-induced ASCs were labeled with 3, 3, 3', 3'-tetramethylindo-carbocyanine perchlorate (DiI), a fluorescent marker, in vitro to be used as seed cells. Then, they were combined with injectable fibrin glue scaffold at 1 x 10(7)/ml cell density. Six athymic BALB/C mice underwent subcutaneous injection of adipogenic-induced ASCs with fibrin glue scaffold at the density of 1 x 10(7) cells/ml into the left side of the low back (induced group), subcutaneous injection of non-adipogenic-induced ASCs into the right side of the low back (non-induced group), and subcutaneous injection of injectable fibrin glue scaffold into the middle part of the neck (blank control group), with 0.2 ml per injection. Twelve weeks later the mice were killed and the implants were taken out. The wet weight was measured. HE and oil red O staining and light and fluorescence microscopy were used for morphological observation.
Adipose tissue-like new-born tissues were found in the injection sites of the induced and un-induced groups. The average wet-weight of the induced group new-born tissue was (28 +/- 15) mg, significantly heavier than that of the un-induced group [(22 +/- 16) mg, P < 0.01]. HE staining and oil red O staining confirmed that the new-born tissue of the induced group was mature adipose tissue and DiI fluorescent staining approved its exogenousness. Most part of the new-born tissues of the un-induced group was fibroid tissue with only a few mature adipose tissues.
ASCs extracted from the lipid part after liposuction can be used as seed cells, mixed, after adipose-induction, with injectable scaffold of fibrin glue, and injected into the body to build mature adipose tissue.
探讨构建组织工程化脂肪组织的可能性，寻找修复软组织缺损的新方法。
采用酶消化法从人抽脂吸出物的脂质部分提取脂肪组织来源干细胞（ASCs），进行培养，部分进行成脂诱导。将成脂诱导及未诱导的ASCs在体外以荧光标记物3,3,3',3'-四甲基吲哚羰花青高氯酸盐（DiI）标记作为种子细胞。然后，将其以1×10(7)/ml的细胞密度与可注射纤维蛋白胶支架复合。6只无胸腺BALB/C小鼠，将成脂诱导的ASCs与纤维蛋白胶支架以1×10(7)细胞/ml的密度皮下注射至左后腰（诱导组），将未诱导的ASCs皮下注射至右后腰（未诱导组），将可注射纤维蛋白胶支架皮下注射至颈部中部（空白对照组），每处注射0.2 ml。12周后处死小鼠，取出植入物，测量湿重。采用苏木精-伊红（HE）染色、油红O染色及光镜和荧光显微镜进行形态学观察。
诱导组和未诱导组注射部位均发现有脂肪组织样新生组织。诱导组新生组织平均湿重为(28±15)mg，明显重于未诱导组[(22±16)mg，P<0.01]。HE染色和油红O染色证实诱导组新生组织为成熟脂肪组织，DiI荧光染色证实其外源性。未诱导组新生组织大部分为纤维组织，仅有少量成熟脂肪组织。
抽脂后脂质部分提取的ASCs可作为种子细胞，经脂肪诱导后与可注射的纤维蛋白胶支架混合并注入体内构建成熟脂肪组织。
