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大鼠耳蜗前体细胞的体外培养与分化鉴定
  • ISSN号:1007-4368
  • 期刊名称:《南京医科大学学报:自然科学版》
  • 时间:0
  • 分类:Q813.1[生物学—生物工程]
  • 作者机构:[1]南京医科大学生物技术系,江苏南京210029, [2]南京医科大学第一附属医院耳鼻咽喉科,江苏南京210029
  • 相关基金:国家自然科学基金(31171217)
中文摘要:

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。

英文摘要:

Objective:We sought to cuhure and isolate cochlear progenitor cells (CPCs),identify their proliferation and differentiation capacity,and demonstrate the expression of genes involved with the development of inner ear during the CPCs differentiation. Methods: CPCs were isolated from newborn rats' cochlea,in vitro cultured and differentiated by serum. Proliferation and differentiation of hair cells were identified by immunocytochemistry method. The expressions of Sox2,CyclinA2,and Jagged1 genes during CPCs differentiation were analyzed using real time qRT-PCR. Results:The cultured CPCs expressed nestin and BrdU, indicating the proliferation ability of CPCs undergoing mitosis. The expression of myosin VII A in the CPCs-derived differentiated cells indicated the CPCs' potential to differentiate into hair cells. It was also noteworthy that the expressions of Sox2 and CyclinA2 were downregulated and the Jaggedl expression was upregulated during the CPCs differentiation. Conclusion:The CPCs in vitro culture showed that it has the ability of proliferation and potential of differentiation into hair ceils. Additionally,it was suggested that Sox2,CyclinA2 and Jagged 1 genes could potentially participate in the modulation of CPCs differentiation and development of cochlear tissues in rats.

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期刊信息
  • 《南京医科大学学报:自然科学版》
  • 中国科技核心期刊
  • 主管单位:
  • 主办单位:南京医科大学
  • 主编:沈洪兵
  • 地址:南京市龙眠大道101号
  • 邮编:211166
  • 邮箱:nyxb@njmu.edu.cn
  • 电话:025-86869293 86869297
  • 国际标准刊号:ISSN:1007-4368
  • 国内统一刊号:ISSN:32-1442/R
  • 邮发代号:28-61
  • 获奖情况:
  • 中国期刊方阵“双效”期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),波兰哥白尼索引,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:18896