软骨源形态发生蛋白对残耳软骨细胞体外增殖的实验研究

The experimental study on the effect of CDMP-1 on proliferation of residual ear chondrocytes of microtia cultured in vitro

目的研究残耳软骨细胞在体外扩增时生物学特性的变化，以及软骨源形态发生蛋白（cartilage-derived morphogenetic protein-1，CDMP-1）对残耳软骨细胞生长分化的影响。方法在体外培养残耳软骨细胞，通过Ⅱ型胶原免疫组织化学染色和甲苯胺兰染色方法，研究CDMP-1对残耳软骨细胞分化的影响；通过MTT法在酶标仪上测定CDMP-1对残耳软骨细胞增殖的影响；使用流式细胞仪测定CDMP-1对残耳软骨细胞周期的影响。结果残耳软骨细胞在体外培养时随代数增加呈异染性的糖胺多糖减少，加入CDMP-1实验组Ⅱ型胶原阳性的比例明显高于对照组（P〈0．05），并且当CDMP-1的浓度为100ng／ml时促细胞增殖效应最大。结论CDMP-1对体外培养的残耳软骨细胞有促进增殖和维持细胞表型的作用。

Objective To investigate the alternation of biological characteristics of residual ear ehondrocytes when proliferating in vitro, and study the effects of CDMP-1 on proliferation and differentiation of residual ear ehondrocytes. Methods Residual ear chondrocytes were cultured in vitro. Then we observed the effect of CDMP-1 on differentiation by immunohistochemistry of collagen type Ⅱ and dyeing with toluidine blue, investigated proliferation effects of CDMP-1 by method of MTT, and analysisd cell cycle changes by flow cytometry. Results The quantity of GAG gradually decreased along with the increase of the doubling time of ehondrecytes in vitro. The quantity of chondrocytes with positive expression of collagen type Ⅱ were significantly more in the experimental group used with CDMP-1 than the contrast. And the most efficient concentration of CDMP-1 was 100 ng/ml. Conclusions CDMP-1 had a good effect on proliferating and could maintain morphology of the residual ear chondrecytes of microtia cultured in vitro.