Chen Guoping, Liu Dechang, Tadokoro Mika, Hirochika Rei, Ohgushi Hajime, Tanaka Junzo, Tateishi Tetsuya
Biomaterials Center, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan.
Biochem Biophys Res Commun. 2004 Sep 10;322(1):50-5. doi: 10.1016/j.bbrc.2004.07.071.
Human mesenchymal stem cells (MSCs) were cultured in vitro in a cobweb-like biodegradable polymer scaffold: a poly(dl-lactic-co-glycolic acid)-collagen hybrid mesh in serum-free DMEM containing TGF-beta3 for 1-10 weeks. The cells adhered to the hybrid mesh, distributed evenly, and proliferated to fill the spaces in the scaffold. The ability of the cells to express gene encoding type I collagen decreased, whereas its ability to express type II collagen and aggrecan increased. Histological examination by HE staining indicated that the cells showed fibroblast morphology at the early stage and became round after culture for 4 weeks. The cartilaginous matrices were positively stained by safranin O and toluidine blue. Immunostaining with anti-type II collagen and anti-cartilage proteoglycan showed that type II collagen and cartilage proteoglycan were detected around the cells. In addition, a homogeneous distribution of cartilaginous extracellular matrices was detected around the cells. These results suggest the chondrogenic differentiation of the mesenchymal stem cells in the hybrid mesh. The PLGA-collagen hybrid mesh enabled the aggregation of mesenchymal stem cells and provided a promotive microenvironment for the chondrogenic differentiation of the MSCs.
人骨髓间充质干细胞（MSCs）在体外培养于蛛网状可生物降解聚合物支架中：一种聚（dl-乳酸-共-乙醇酸）-胶原蛋白混合网，置于含转化生长因子-β3的无血清杜氏改良 Eagle 培养基（DMEM）中培养 1 - 10 周。细胞附着于混合网，分布均匀，并增殖以填充支架中的间隙。细胞表达 I 型胶原蛋白编码基因的能力下降，而其表达 II 型胶原蛋白和聚集蛋白聚糖的能力增加。苏木精-伊红（HE）染色的组织学检查表明，细胞在早期呈成纤维细胞形态，培养 4 周后变为圆形。软骨基质经番红 O 和甲苯胺蓝染色呈阳性。用抗 II 型胶原蛋白和抗软骨蛋白聚糖进行免疫染色显示，在细胞周围检测到 II 型胶原蛋白和软骨蛋白聚糖。此外，在细胞周围检测到软骨细胞外基质的均匀分布。这些结果表明间充质干细胞在混合网中发生软骨分化。聚乳酸-乙醇酸共聚物-胶原蛋白混合网能够使间充质干细胞聚集，并为骨髓间充质干细胞的软骨分化提供促进性微环境。
