论文部分内容阅读
目的 :分离、培养大鼠的耳蜗前体细胞,鉴定其增殖和分化能力,分析内耳发育相关基因在其分化中的表达。方法 :从新生大鼠的耳蜗组织中分离获得前体细胞,进行体外培养,并加入血清诱导分化,通过免疫细胞化学的方法鉴定其增殖和分化为毛细胞的能力,利用荧光定量PCR的方法分析不同分化阶段的耳蜗前体细胞中内耳发育相关基因的表达。结果:原代培养的耳蜗前体细胞经免疫细胞化学鉴定呈nestin、Brd U阳性,经血清诱导分化后呈myosinⅦA阳性。荧光定量PCR的结果表明Sox2、Cyclin A2在耳蜗前体细胞分化的过程中表达逐渐下降,而Jagged1在分化过程中表达不断升高。结论 :分离的耳蜗前体细胞具有增殖能力和分化为毛细胞的潜能,在其分化过程中Sox2、Cyclin A2和Jagged1可能参与调控大鼠前体细胞的分化和耳蜗组织的发育。
OBJECTIVE: To isolate and culture rat cochlear progenitor cells, identify their proliferation and differentiation ability, and analyze the expression of inner ear development-related genes in their differentiation. Methods: The precursor cells were isolated from the cochlea of neonatal rats, cultured in vitro and induced to differentiate by adding serum. The ability of proliferating and differentiating into hair cells was identified by immunocytochemistry, and analyzed by real-time quantitative PCR Expression of inner ear development-related genes in different stages of differentiation of cochlear progenitor cells. Results: Primary cultured cochlear progenitor cells were identified by immunocytochemistry as nestin and BrdU positive. After being differentiated by serum, myosinⅦA positive. Fluorescent quantitative PCR results showed that the expression of Sox2 and Cyclin A2 gradually decreased in the process of cochlear progenitor cell differentiation, while the expression of Jagged1 increased during differentiation. CONCLUSIONS: Isolated cochlear progenitor cells have the potential to proliferate and differentiate into hair cells. Sox2, Cyclin A2 and Jagged1 may be involved in the regulation of rat precursor cell differentiation and cochlear tissue development during their differentiation.