脂肪源性干细胞与胶原海绵支架的相容性

摘要

Objective:To observe the growth of adipose-derived stem cells (ADSC) which seeded in the collagen sponge scaffold and study the compatibility of ADSC and collagen sponge. Methods: The inguinal adipose tissues isolated from SD rats were digested by collagenase I. After culture, purification and passage, the 3rd generation ADSC were collected to observe and analyze the cell growth curve. The surface markers of the 3rd generation cells were stained by immunofluorescence. The 3rd generation ADSC were stained by CM-Dil and seeded into the collagen sponges. The construsts were observed under a fluorescence microscope. The sections stained with HE of the construsts were observed by microscopy to assess the compatibility of the ADSC and collagen sponge. Results: ADSC showed a spindle or triangle shape and were arranged in a whirl-like form and in a certain oritentation. The passage cells growth curves had common characteristics: latent period was 24-48 hours, logarithm multiplication period was 3-4 days, and the cells entered the platform period at 5-6 days after the logarithm multiplication period. The immunofluorescence staining of the 3rd generation of ADSC showed the expressions of CD13, CD29 and CD44. The adipose-derived stem cells proliferated well in collagen sponges. The number of ADSC in collagen sponges did not increase at day 3, compared with that at day 1. The number of cells in collagen sponges was significantly more at day 5 and 7 than that at day 1 and 3. Conclusion: Collagen sponge had a good compatibility with ADSC. The construction of adipose-derived stem cells and collagen sponges can be used in the tissue engineering research and applicatioa%目的:研究大鼠脂肪源性干细胞(ADSC)在胶原海绵支架上生长情况,探讨ADSC和胶原海绵的相容性.方法:取SD大鼠腹股沟处的脂肪组织,经Ⅰ型胶原酶消化离心后分离培养大鼠ADSC,观察分析第3代细胞的生长曲线,采用细胞免疫荧光法检测第3代细胞表面分子标志的表达,将第3代细胞种植到胶原海绵支架上,采用CM-Dil特异性的细胞膜染色和切片H-E染色,评价大鼠ADSC与胶原海绵的相容性.结果:ADSC呈长梭形或多角形,细胞排列有一定的方向性,呈漩涡状生长.第3代细胞在第4、5天进入对数生长期,第7天到达平台期.细胞免疫荧光显色显示CD13、CD29、CD44均为阳性.种植在胶原海绵支架上体外培养,细胞存活,沿胶原支架表面和胶原海绵空洞生长,复合体体外培养的前3d细胞数量没有明显的变化,但第5天和第7天细胞数量发生明显的增加.结论:大鼠ADSC与胶原海绵具有很好的相容性,ADSC和胶原海绵的构建物可应用于组织工程的研究与应用.