CAJ | 학술논문

背景：现有研究表明，人与大鼠卵巢优势卵泡中的颗粒细胞具有表达干细胞特性的现象。目的：探讨干细胞相关因子在大鼠卵巢颗粒细胞中的表达。方法：将大鼠卵巢组织制作石蜡切片后，使用免疫组化法检测CD34，CD133，ABCG2/Bcrp1，Pou5f1/Oct-4的表达。卵泡穿刺法获取颗粒细胞，并使用免疫组化法检测FSHR受体的表达以鉴定颗粒细胞的纯度。免疫组化法检测颗粒细胞 CD44， C-Kit 的表达情况， RT-PCR 检测卵巢组织与颗粒细胞 ABCG2/Bcrp1、Pou5f1/Oct-4、Nanog基因的表达情况。结果与结论：免疫组化法石蜡切片显示部分卵巢颗粒细胞CD34，CD133，ABCG2/Bcrp1，Pou5f1/Oct-4阳性表达，而Pou5f1/Oct-4蛋白的表达在卵泡发育的周期中逐步增多，并在黄体化的时候显著增强，形成白体后则消失，因而具有周期性表达的特点。卵泡穿刺法提取的原代颗粒细胞FSHR受体鉴定阳性率在95%以上。体外培养的颗粒细胞以长梭形或菱形为主，部分细胞有聚集生长现象，且 CD44，C-Kit 阳性表达。RT-PCR检测结果显示，卵巢组织与体外培养的颗粒细胞均不表达 Nanog，卵巢低表达 Pou5f1/Oct-4，强烈表达ABCG2/Bcrp1，颗粒细胞微弱表达Pou5f1/Oct-4，而ABCG2/Bcrp1表达较强。以上结果显示大鼠卵巢中的部分颗粒细胞具有干细胞的特性。
배경：현유연구표명，인여대서란소우세란포중적과립세포구유표체간세포특성적현상。목적：탐토간세포상관인자재대서란소과립세포중적표체。방법：장대서란소조직제작석사절편후，사용면역조화법검측CD34，CD133，ABCG2/Bcrp1，Pou5f1/Oct-4적표체。란포천자법획취과립세포，병사용면역조화법검측FSHR수체적표체이감정과립세포적순도。면역조화법검측과립세포 CD44， C-Kit 적표체정황， RT-PCR 검측란소조직여과립세포 ABCG2/Bcrp1、Pou5f1/Oct-4、Nanog기인적표체정황。결과여결론：면역조화법석사절편현시부분란소과립세포CD34，CD133，ABCG2/Bcrp1，Pou5f1/Oct-4양성표체，이Pou5f1/Oct-4단백적표체재란포발육적주기중축보증다，병재황체화적시후현저증강，형성백체후칙소실，인이구유주기성표체적특점。란포천자법제취적원대과립세포FSHR수체감정양성솔재95%이상。체외배양적과립세포이장사형혹릉형위주，부분세포유취집생장현상，차 CD44，C-Kit 양성표체。RT-PCR검측결과현시，란소조직여체외배양적과립세포균불표체 Nanog，란소저표체 Pou5f1/Oct-4，강렬표체ABCG2/Bcrp1，과립세포미약표체Pou5f1/Oct-4，이ABCG2/Bcrp1표체교강。이상결과현시대서란소중적부분과립세포구유간세포적특성。
BACKGROUND:Human and rat ovarian granulosa cels in dominant folicles have the phenomenon of expressing stem cel characteristics. OBJECTIVE:To investigate the expression of stem cel-related factors in rat ovarian granulosa cels. METHODS: After the paraffin sections of rat ovarian tissue, immunohistochemical method was used to detect CD34, CD133, ABCG2/Bcrp1, Pou5f1/Oct-4 expressions. Granulosa cels culturedin vitro were harvested by folicular puncture method, and then the immunohistochemical method was used to detect the expression of FSHR receptor in order to identify the purity of granule cels. In the cultured granulosa cels, CD44 and C-Kit expressions were detected immunohistochemicaly, RT-PCR was used to detect ABCG2/Bcrp1, Pou5f1/Oct-4, Nanog gene expressions in ovarian tissue and granulosa cels. RESULTS AND CONCLUSION:Immunohistochemistry detection on paraffin sections showed that a part of ovarian granulosa cels expressed CD34, CD133, ABCG2/Bcrp1 and Pou5f1/Oct-4, and the expression of Pou5f1/Oct-4 protein gradualy increased in the development of ovarian folicles, significantly enhanced during the luteal phase, and then disappeared after the formation of corpus albicans, displaying a periodic expression characteristics. FSHR receptor positive identification rate of primary cels harvested by the foliclar puncture method was more than 95%. Granulosa cels culturedin vitrowere mainly long spindle-shaped or diamond, and some cels presented with aggregation growth and expressed CD44 and C-Kit. RT-PCR test results showed that there were no Nanog in the ovarian tissue and cultured granulosa cels, low expression of Pou5f1/Oct-4 in the ovarian tissue, strong expression of ABCG2/Bcrp1 in the ovarian tissue, weak expression of Pou5f1/Oct-4 in the cultured granulosa cels, and strong expression of ABCG2/Bcrp1 in the cultured granulosa cels. These findings suggest that a part of granulosa cels in the rat ovarian have the characteristics of stem cels.