CAJ | 학술논문

目的观察在体外Transwell系统中共培养后,乳房脂肪来源干细胞(human breast adipose-derived stem cells,hbASCs)向乳腺上皮样细胞转化的能力.方法自乳房缩小整形术切除的腺体内的脂肪组织中获取hbASCs,取HBL-100细胞系和第3代hbASCs,按1∶1的细胞比例分别接种于Transwell共培养系统的上室和下室,同时设置相同浓度单纯第3代hbASCs为对照组,分别在DMEM/F12培养基中培养,于相差显微镜下观察实验和对照组下室细胞形态变化及生长情况,透射电子显微镜下观察下室细胞超微结构的改变,并对细胞爬片进行荧光免疫细胞化学染色,以鉴定乳腺上皮细胞特异性标志物CK18、CK19的表达.结果 Transwell体系中各细胞均可良好贴壁生长.共培养至15 d,实验组下室细胞数量较对照组明显增多,体积变小,排列呈铺路石样,部分细胞形态上表现出乳腺上皮样细胞特征;电镜下可见微绒毛、桥粒和张力丝等典型上皮细胞结构特征;免疫荧光结果显示,经共培养诱导后,实验组部分细胞可阳性表达乳腺上皮细胞特异性标记物CK18、CK19;对照组诱导细胞上述检测结果均为阴性.实验组CK18、CK19阳性率分别为(24.4±12.0)％和(21.6±16.4)％;对照组分别为(1.8±1.7)％和(1.1±0.6)％,2组比较差异有统计学意义(P＜0.01).结论与HBL-100细胞共培养后,乳房脂肪干细胞可以被成功诱导成乳腺上皮样细胞.
목적 관찰재체외Transwell계통중공배양후,유방지방래원간세포(human breast adipose-derived stem cells,hbASCs)향유선상피양세포전화적능력.방법 자유방축소정형술절제적선체내적지방조직중획취hbASCs,취HBL-100세포계화제3대hbASCs,안1∶1적세포비례분별접충우Transwell공배양계통적상실화하실,동시설치상동농도단순제3대hbASCs위대조조,분별재DMEM/F12배양기중배양,우상차현미경하관찰실험화대조조하실세포형태변화급생장정황,투사전자현미경하관찰하실세포초미결구적개변,병대세포파편진행형광면역세포화학염색,이감정유선상피세포특이성표지물CK18、CK19적표체.결과 Transwell체계중각세포균가량호첩벽생장.공배양지15 d,실험조하실세포수량교대조조명현증다,체적변소,배렬정포로석양,부분세포형태상표현출유선상피양세포특정;전경하가견미융모、교립화장력사등전형상피세포결구특정;면역형광결과현시,경공배양유도후,실험조부분세포가양성표체유선상피세포특이성표기물CK18、CK19;대조조유도세포상술검측결과균위음성.실험조CK18、CK19양성솔분별위(24.4±12.0)％화(21.6±16.4)％;대조조분별위(1.8±1.7)％화(1.1±0.6)％,2조비교차이유통계학의의(P＜0.01).결론 여HBL-100세포공배양후,유방지방간세포가이피성공유도성유선상피양세포.
Objective To explore the feasibility of the transdiferentiation of human breast adiposederived stem cells (hbASCs) into mammary epithelial-like cells after co-culturing in Transwell in vitro.Methods The third passage hbASC and the HBL-100 cell line were co-cultured in a Transwell culture system for 15 days.The hbASCs were observed and identified by invcrted phase contrast microscope and transmission electron microscopy,and immunocytochemistry staining in the induced and control groups.Results Both the third passage hbASCs and the HBL-100 cell line cells could adhere and grow rapidly after co-culture in the Transwell system.After co-culture for 15 days,the morphology of some induced hbASCs changed into epithelial-like cells.Some induced hbASCs showed positive expression of CK18,CK19 by immunocytochemistry staining,and typical epithelium cells with microvilli,desmosomes and tonofilaments observed under TEM.The positive rate of CK18 and CK19 was (24.4 ± 12.0)％ and (21.6 ± 16.4)％ in experimental group,and(1.8 ± 1.7)％ and(1.1 ±0.6)％ in control group.Conclusion The data suggests that hbASCs may have the potential to transdifferentiate into human mammary epithelial-like cells after co-culturing in Transwell in vitro.