诱导性多潜能干细胞向卵巢颗粒细胞定向分化的体外研究  

In Vitra Study on Committed Differentiation of Induced Pluripotent Stem Cells into Ovarian Granulosa Cells

作　　者：韩梦欣 郭路 刘晓程 陈秀英 顾超 李斌 Han Mengxin;Guo Lu;Liu Xiaocheng;Chen Xiuying;Gu Chao;Li Bin(Department of Gynecology,Obstetrics＆ Gynecology Hospital of Fudan University,Shanghai,200090,P.R.China)

机构地区：[1]复旦大学附属妇产科医院妇科,上海200090

出　　处：《老年医学与保健》2018年第3期332-335,共4页Geriatrics & Health Care

基　　金：国家自然科学基金(81471423);上海市自然科学基金(14ZR1404200)

摘　　要：目的体外研究诱导性多潜能干细胞(induced pluripotent stem cell,i PSCs)向卵巢颗粒细胞(Granulosa cell,GC)定向分化。方法利用transwell间接共培养体系诱导i PSCs向GC定向分化。倒置荧光显微镜观察诱导后细胞形态特征;酶联免疫吸附法(Enzyme-linked immunosorbent assay,ELISA)检测共培养上清E2水平;RT-PCR鉴定诱导后细胞颗粒细胞相关基因表达;细胞免疫化学鉴定诱导后细胞FSHR表达。结果诱导后细胞成角形或梭形,与颗粒细胞形态相似;共培养第1、3、5和7天的E2值依次为(35.46±5.68)pg/m L、(44.58±4.10)pg/m L、(50.08±8.20)pg/m L和(63.49±7.04)pg/m L,呈递增趋势,差异具有统计学意义(P<0.05);诱导后细胞FSHR、LHR、AMHR2、FOXL2和CYP19A1基因表达均上调,与i PSCs组相比差异均有统计学意义(P<0.05);诱导后细胞FSHR表达阳性。结论 i PSCs与GC共培养可分化为具有颗粒细胞功能的颗粒样细胞。Objective To perform an in vitra study on committed differentiation of induced pluripotent stem cells（i PSCs）into ovarian granulosa cells（GC）. Methods i PSCs were differentiated into ovarian GC with Transwell co-culture system; the morphological characteristics of the induced cells were observed under Inverted fluorescence Microscope; the E2 concentration of co-culture supernatant was detected by ELISA; RT-PCR was applied in identifying the gene expression of cells after induction and cell immunochemistry in identifying the cell FSHR expression. Results After induction, the cell shape was similar to the morphology of granulosa cells, flat with 3 or more sides or looked like fusiform cells; the E2 concentration at the time point of the 1 st, 3 rd, 5 th and 7 th day of the co-culture supernatant was（35.46±5.68）pg/m L,（44.58±4.10）pg/m L,（50.08±8.2）pg/m L and（ 63.49±7.04）pg/m L, respectively, showing out an increasing trend with statistical difference（P 0.05）; after induction, the expressions of FSHR, LHR, AMHR2, FOXL2 and CYP19 A1 genes were up-regulated, statistically different from those in i PSCs group（ P0.05）; meanwhile, the expression of FSHR was positive after induction. Conclusions i PSCs can successfully differentiate into granulosa-like cells.

关 键 词：诱导性多潜能干细胞 卵巢颗粒细胞 共培养 细胞分化 

分 类 号：R711.75[医药卫生—妇产科学]