大鼠耳蜗毛细胞前体细胞与耳蜗间质细胞共培养的实验研究

Co-culture of Hair Cell Progenitors and Mesenchymal Cells from Rat Cochlea

目的探讨耳蜗间质细胞对耳蜗毛细胞前体细胞——大上皮嵴(greater epithelial ridge,GER)细胞的诱导分化作用。方法取出生后第1天大鼠20只,随机分为2组,每组各10只,利用酶消化和机械分离相结合的方法分别分离出纯耳蜗GER细胞和耳蜗间质细胞,并进行体外混合培养及形态学的观测,对共培养10天的标本行抗Calretinin(标记幼稚毛细胞)及抗p27抗体(标记支持细胞)的免疫细胞组织化学染色鉴定。结果混合培养的耳蜗GER细胞和间质细胞易形成片状细胞岛,共培养10天后,有部分细胞表达Calretinin,亦有少数细胞表达p27,但无Calretinin及p27双标记者。结论GER细胞与耳蜗间质细胞共培养,可以诱导GER细胞分化为毛细胞及支持细胞表型。

Objective To investigate the effect of co - culture with mesenchymal cells on greater epithelial ridge （GER） cells which are likely hair cell progenitors from rat cochlea. Methods Pure GER cells and mesenchyrnal cells were isolated respectively using a combinatorial approach of enzymatic digestion and mechanical separation from P1 rat cochlear and co - cultured in vitro. Immunostaining of anti - calretinin and anti - p27 antibodies were performed 10 days after co - culture. Results Many GER cells after co - cultured with mesenchymal cells were marked by calretinin and p27. Conclusion GER cells have the potential to differentiate into hair cell and support cell phenotype when mixed with dissociated mesenchymal cells prepared from rat cochlear.