摘要
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。
Purpose: To observe the morphological change of in vitro growth of preantral follicle seperated from ovaries of prepuberal mice, and to assess Gn, ITS and EGF's impact on their development. Methods: Mechanical isolation and selection of early preantral mouse follicles (90-130 tan in diameter) from 2 weeks old mice. Follicles were cultured in a medium with or without Gn and individual on microdrop for up to 14 days, change medium and observe everyday, record GC prolification, antral formation and oocyte's maturaion. Results: The medium with Gn supported preantral follicle culture after isolation, during which they retained a three-dimensional structure, maintained oocyte viability and increased in diameter and number of somatic cells. Preantral follicle cultured in Gn medium grew obviously, while those without Gn grew slowly and after 6d's culture began to shrink and blacken. Significant increase in survival rate and maturation rate of oocytes were observed in Gn group (P<0.01), with 92.92% survived and 28.68% 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 IVM, but need to be added with rLH/rFSH, rHCG/rEGF to facilitate membrane cell adhesion, GC proliferation and oocyte maturation.
出处
《复旦学报（医学版）》
EI
CAS
CSCD
北大核心
2004年第1期42-45,共4页
Fudan University Journal of Medical Sciences
基金
上海市自然科学基金(01ZB14037)
