目的 探讨体外定向诱导大鼠骨髓间充质干细胞向耳蜗毛细胞样细胞分化的可行性.方法 分离培养大鼠骨髓间充质干细胞,并加以鉴定.分离出生1~3 d的乳鼠耳蜗Corti器,与骨髓间充质干细胞在体外共培养14 d.通过反转录聚合酶链反应(RT-PCR)和免疫细胞化学染色对分化细胞的特异性分子指标(myosinⅦa、math1及calretinin)进行鉴定.结果 免疫细胞化学染色显示诱导分化后的细胞myosinⅦa、math1和calretinin表达阳性,RT-PCR提示分化后的细胞可表达毛细胞的标志物myosinⅦa和math1.结论 体外培养的骨髓间充质干细胞可诱导分化为具有毛细胞分子标志物的毛细胞样细胞.
Objective To investigate the feasibility of orientation-induced differentiation of rat bone marrow-derived mesenchymal stem cells (MSC) into cochlear hair cell-like cells in vitro. Methods MSC were extracted, cultured and identified. The organ of Corti was isolated from post natal day 1-day 3 SpragueDawley rats. Then MSC were co-cultured with organ of Corti for 14 days in vitro. The expression of hair cell markers (myosinⅦa, math1 and calretinin) was detected by immunocytochemical analyses and by reverse transcription-polymerase chain reaction (RT-PCR). Results The differentiated cells were immunopositive for myosinⅦa, calretinin and math1. And RT-PCR results indicated that these differentiated cells expressed hair cell markers Math1 and myosinⅦa. Conclusions MSC cultured in vitro could be successfully induced to differentiate into hair cell-like cells with hair cell molecular markers.