中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2011年
12期
896-899
,共4页
林盛%龙海霞%向橦%朱波%谢荣凯
林盛%龍海霞%嚮橦%硃波%謝榮凱
림성%룡해하%향동%주파%사영개
肿瘤干细胞%卵巢肿瘤%细胞分离
腫瘤榦細胞%卵巢腫瘤%細胞分離
종류간세포%란소종류%세포분리
Tumor stem cells%Ovarian neoplasms%Cell separation
目的 从人卵巢癌组织中分离并鉴定卵巢癌干细胞.方法 取新鲜卵巢癌组织,以胶原酶和透明质酸酶充分消化,获得单细胞悬液,经红细胞裂解液处理,CD133磁珠孵育进行磁性细胞分选,流式细胞仪验证分选效率.将阳性细胞置于含人表皮生长因子、碱性成纤维细胞生长因子、牛血清白蛋白和人胰岛素的无血清培养基中,待细胞成球状生长后进行免疫荧光、分化实验和成瘤实验,鉴定其干细胞特性.结果 从人卵巢癌组织中成功分离出肿瘤干细胞,流式细胞仪检测其CD133的表达率为82.5％.该细胞在无血清培养基中呈悬浮球状生长,具有自我更新能力、分化潜能和更强的致瘤能力,具备干细胞特性.结论 人卵巢癌组织中存在卵巢癌干细胞,利用CD133磁珠分选得到的肿瘤干细胞具备干细胞的相关特性,可用于后续卵巢癌干细胞生物学特性的研究.
目的 從人卵巢癌組織中分離併鑒定卵巢癌榦細胞.方法 取新鮮卵巢癌組織,以膠原酶和透明質痠酶充分消化,穫得單細胞懸液,經紅細胞裂解液處理,CD133磁珠孵育進行磁性細胞分選,流式細胞儀驗證分選效率.將暘性細胞置于含人錶皮生長因子、堿性成纖維細胞生長因子、牛血清白蛋白和人胰島素的無血清培養基中,待細胞成毬狀生長後進行免疫熒光、分化實驗和成瘤實驗,鑒定其榦細胞特性.結果 從人卵巢癌組織中成功分離齣腫瘤榦細胞,流式細胞儀檢測其CD133的錶達率為82.5％.該細胞在無血清培養基中呈懸浮毬狀生長,具有自我更新能力、分化潛能和更彊的緻瘤能力,具備榦細胞特性.結論 人卵巢癌組織中存在卵巢癌榦細胞,利用CD133磁珠分選得到的腫瘤榦細胞具備榦細胞的相關特性,可用于後續卵巢癌榦細胞生物學特性的研究.
목적 종인란소암조직중분리병감정란소암간세포.방법 취신선란소암조직,이효원매화투명질산매충분소화,획득단세포현액,경홍세포렬해액처리,CD133자주부육진행자성세포분선,류식세포의험증분선효솔.장양성세포치우함인표피생장인자、감성성섬유세포생장인자、우혈청백단백화인이도소적무혈청배양기중,대세포성구상생장후진행면역형광、분화실험화성류실험,감정기간세포특성.결과 종인란소암조직중성공분리출종류간세포,류식세포의검측기CD133적표체솔위82.5％.해세포재무혈청배양기중정현부구상생장,구유자아경신능력、분화잠능화경강적치류능력,구비간세포특성.결론 인란소암조직중존재란소암간세포,이용CD133자주분선득도적종류간세포구비간세포적상관특성,가용우후속란소암간세포생물학특성적연구.
Objective To isolate and identify the cancer stem cells from primary human ovarian cancer tissues.Methods Fresh tumor tissues from five cases of pathologically diagnosed ovarian cancers were taken,minced and then digested with collagenase and hyaluronidase to obtain single cell suspension.The erythrocytes were removed with ACK Lysis buffer.The suspensions were sorted by magnetic activated cell sorting (MACS) using CD133-binding microbeads.Then the sorted CD133 + cells were verified by flow cytometry.The cells were cultured in serum-free medium supplemented with EGF,bFGF,insulin and BSA,and grew into spheroids.Immunofluorescence,differentiation and tumor formation tests of the cells were performed to characterize the properties of cancer stem cells.Results The ovarian cancer stem cells were successfully isolated from primary human ovarian tumors,which formed typical spheroids in serum-free medium and had stronger ability of tumorigenesis.The results of related experiments verified that CD133positive cells owned the properties of cancer stem cells. Conclusions The ovarian cancer stem cells presenting the characteristics of sternness in vitro and in vivo,have been successfully isolated from primary human ovarian tumor tissues by MACS.The isolated ovarian cancer stem cells could be used in future researches of their biological functions.
