重庆医学2017，Vol.46Issue(13)：1792-1796,5.DOI:10.3969/j.issn.1671-8348.2017.13.021
大鼠卵巢颗粒细胞体外培养及哈蟆油对其增殖功能的影响
Culture in vitro of rat ovarian granulosa cells and effect of Ranae Oviductus on its proliferation
摘要
Abstract
Objective To optimize the effective methods of isolation,purification,culture in vitro and identification of SD rat ovarian granulosa cells,to research the effects of Oviductus Ranae on the proliferation of rat ovarian granulosa cells by CCK-8,and to contrastively analyze the best optimal action concentration and time of serum contsining Oviductus Ranae on granulosa cells to lay the foundation for further in vitro experiment.Methods Nonage SD rats aged 25 d were selected and intraperitoneally injected by pregnant mare serum,then killed after 48 h.Ovarian granulosa cells were collected and cultured in the DMEM-F12 culture solution.The hematoxylin & eosin(HE) staining and immunofluorescence technique were used to identify the ovarian granulosa cells.Twen ty-five SD rats were randomly divided into the normal control group,positive medicine control group,and low,middle and high do ses Oviductus Ranae groups.Blood was collected and serum was separated after 7 d mediaction gavage.The volume percent of 10 ％,20％,40％,80％ serum in each group was added into the in vitro medium system of ovarian granulosa cells culture.Then the cell proliferation situation at 24,48,72 h in each group was measured by CCK-8.Results Oviductus Ranae significantly increased the proliferation ability of granulosa cells in a certain dose-dependent relation.With the increase of Oviductus Ranae concentration con centration,its.proliferation ability was gradually increased,after 48 h action,which in the Oviducthus Ranae-Comtaining serum group with the volume fraction of 20％ was most significant (P＜0.05).Conclusion Establishing in vitro cultural method of rat o varian granulosa cells is conductive to further research the action and mechanism of Oviductus Ranae on ovary.关键词
哈蟆油/卵巢颗粒细胞/增殖/含药血清/CCK-8Key words
Oviductus Ranae/ovarian granulosa cells/proliferation/drug-containing serum/CCK-8分类
医药卫生引用本文复制引用
零小妹,谭焱,季波,杨晶晶,吴新荣,梁磊..大鼠卵巢颗粒细胞体外培养及哈蟆油对其增殖功能的影响[J].重庆医学,2017,46(13):1792-1796,5.基金项目
广东省广州市珠江科技新星项目(2013J2200030) （2013J2200030）
广东省广州市科技计划项目(201509010012). （201509010012）
