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上海市自然科学基金(01ZB14037)

作品数:2 被引量:5H指数:1
相关作者:吴晓云陈淑颖顾敦瑜高敏芝陆湘更多>>
相关机构:上海市计划生育科学研究所更多>>
发文基金:上海市自然科学基金更多>>
相关领域:医药卫生生物学更多>>

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In-vitro Maturation of Immature Oocytes from Preantral Follicles in Prepuberal Mice
2007年
Objective To observe the morphological changes in in vitro growth of preantral follicle isolated from prepuberal mice and to assess impacts of gonadotropin (Gn), insulin transferrin selenium (ITS) and epidermal growth factor (EGF) on their development. Methods Early preantral mice follicles (90-130μm diameter) were mechanical isolated and selected from 2 weeks' old mice and then cultured in alpha-minimal essential medium (α-MEM) with or without Gn, ITS and EGF. The preantral follicles were cultured singly in 20 miovliters droplets for up to 14 d. The medium was replaced and the .follicles were observed everyday. Granulosa cells (GC) prolification, antrum formation and oocyte maturation were recorded. Results The medium with Gn supported preantral follicle culture in vitro, during which they retained a three-dimensional structure, maintained oocytes viability and increased in diameter and number of somatic cells'. Preantral follicles cultured in Gn medium grew obviously, while those without Gn grew slowly and after 6 d's culture began to shrink and blacken. Significant increase in survival rate and maturation rate of oocytes was observed in Gn group (P〈0.01), with 92.9% survived and 28.7%formed an antrum. Further supplementation of the Gn medium with ITS and rLH, resulted in the significant increase in survival and maturation of preantral follicle (P〈0.05) Conclusions α-MEM can be the medium for in vitro culture (IVC) of preantral follicles, but need to be added with rLH/rFSH, rHCG/rEGF to facilitate thecal cell attachment, GC proliferation and oocyte maturation.
Min-zhi GAO Yu-bao WANG Xiao-yun WU
小鼠窦前卵泡的体外培养被引量:5
2004年
目的 观察性成熟前小鼠的窦前卵泡体外培养的形态学改变,同时评价激素及生长因子等添加物对窦前卵泡体外发育的影响。方法 机械法分离性成熟前小鼠卵巢中的窦前卵泡,分2组培养于20μL培养液滴内。(1)Gn组:α-MEM+10%FCS+100 mU/mL rFSH±ITS(5 μg/mL insulin,5 μg/mL transferrine,5 ng/mLselenium)±10 mU/mL rLH,第12天移入 α-MEM+1.5 U/mL rHCG+5 ng/mL rEGF+10% FCS。(2)nGn组:α-MEM+10% FCS。每天换液观察,记录GC增生、窦形成等形态学改变和卵母细胞成熟情况。结果 激素组窦前卵泡增长明显,92.92%存活,28.65%形成窦,培养14d后40.35%排出成熟卵;而无激素组窦前卵泡生长缓慢,生存6d后开始萎缩、变黑,存活率仅10.91%,无窦卵泡形成,仅11.11%排出成熟卵,差异有显著意义(P<0.01)。激素组在添加ITS、LH后,窦前卵泡的窦形成率和排出成熟卵母细胞率均显著上升(P<0.05)。结论 α-MEM可作为窦前卵泡体外培养液,但必须添加rLH/rFSH、ITS、rHCG/rEGF以促进膜细胞黏附、颗粒细胞增生和卵母细胞成熟。
汪玉宝高敏芝顾敦瑜吴晓云陈淑颖陆湘何志英
关键词:小鼠窦前卵泡体外培养形态学卵泡发育
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