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家兔子宫平滑肌细胞的体外分离培养
毕台飞1,2, 宋宇轩3, 窦 铖4
1.榆林学院 生命科学学院;2.陕西省绒山羊工程技术研究中心;3.西北农林科技大学 动物科技学院;4.北京中地种畜股份有限公司
摘要:
【目的】研究家兔子宫平滑肌层细胞的体外培养和扩增方法。【方法】应用酶解消化法对新西兰大白兔的子宫平滑肌细胞进行体外分离和培养,并研究不同水平性腺激素对其生长的影响。【结果】酶解消化法可成功进行家兔子宫平滑肌细胞的体外培养和传代。子宫平滑肌细胞在体外的增殖受卵巢性腺激素的影响,100 nmol/L雌二醇对其体外增殖有促进作用,而100 nmol/L孕酮则有抑制作用。免疫组织化学结果表明,体外培养的子宫平滑肌细胞在激素作用下仍可维持其特性,在体外可传9~10代。【结论】建立了家兔子宫平滑肌细胞的体外培养和扩增方法,可为构建子宫三维结构提供充足的种子细胞。
关键词:  家兔  子宫平滑肌细胞  雌二醇  孕酮  体外培养
DOI:
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基金项目:西北农林科技大学博士科研启动基金项目(Z111020721)
Isolation and culture of rabbit uterine myometrial cell in vitro
BI Tai-fei,SONG Yu-xuan,DOU Cheng
Abstract:
【Objective】This study investigated the isolation and culture of uterine myometrial cell in vitro.【Method】In this study,tissue isolation and trypsin digestion method was used to isolate and culture uterine myometrial cell from New Zealand White rabbits in vitro,and effects of β-estradiol (E2) and progesterone (P4) levels on the proliferation of myometrial cells were investigated.【Result】The uterine myometrial cells were successfully prepared by tissue cutting and 0.25% trypsin solution digested.The proliferation of the uterine myometrial cells was largely affected by E2 and P4.100 nmol/L E2 increased the proliferation,while 100 nmol/L P4 inhibited the proliferation (P<0.05).Immunocytochemistry staining demonstrated that the uterine myometrial cells could maintain their behaviors with the help of hormones for 9-10 generations in vitro.【Conclusion】The established culture and proliferation of uterine myometrial cells could provide enough seeding cells for the construction of 3D uterine structure.
Key words:  rabbit  uterine myometrial cell  β-estradiol  progesterone  culture in vitro