CAJ | 학술논문

背景:现阶段对于肝干细胞在胚胎肝发育过程中的形态学特征、时空分布、分化过程的报道很少.目的:以甲胎蛋白、细胞角蛋白19、c-Met作为标志物,观察其在不同发育阶段肝干细胞中的时空分布.设计、时间及地点:细胞学体外观察,于2005-06/2006-12在潍坊医学院中心实验室完成.材料:40例胚胎标本取自3个月内的流产胎儿,由潍坊医学院附属医院提供.方法:收集流产胚胎,30 min内制作切片,显微镜下根据胚层形成情况、体节数目及器官发育状况确定胎龄.选择胎龄第3～12周的标本切片,每隔10张抽出1张切片,行免疫组织化学染色.主要观察指标:第3～12周胚胎肝干细胞甲胎蛋白、c-Met、细胞角蛋白19的表达.结果:第3～5周甲胎蛋白、c-Met呈阳性反应,提示为肝干细胞;第10～12周甲胎蛋白、c-Met阳性细胞主要分布于汇管区周围,提示此时肝干细胞主要局限于汇管区周围的肝索内,与成年肝中卵圆细胞(成年肝干细胞)的分布一致.细胞角蛋白19阳性反应在第7周时开始出现;第10～11周时细胞角蛋白19阳性反应主要位于汇管区附近的肝索细胞和胆管板细胞及胆管上皮细胞;第12周时细胞角蛋白19阳性信号仪见于胆管板和胆管上皮细胞.此时所有的胆管板细胞及胆管上皮细胞均呈甲胎蛋白、c-Met和细胞角蛋白19阳性.结论:细胞角蛋白19在肝干细胞中无表达,仅在胆管上皮细胞及其祖细胞中有表达,可能不适合作为肝干细胞的标志物.所有的胆管板及胆管上皮细胞均呈甲胎蛋白、c-Met、细胞角蛋白19阳性反应,推测甲胎蛋白+/c-Met+/细胞角蛋白19+的细胞可能为胆管祖细胞.
배경:현계단대우간간세포재배태간발육과정중적형태학특정、시공분포、분화과정적보도흔소.목적:이갑태단백、세포각단백19、c-Met작위표지물,관찰기재불동발육계단간간세포중적시공분포.설계、시간급지점:세포학체외관찰,우2005-06/2006-12재유방의학원중심실험실완성.재료:40례배태표본취자3개월내적유산태인,유유방의학원부속의원제공.방법:수집유산배태,30 min내제작절편,현미경하근거배층형성정황、체절수목급기관발육상황학정태령.선택태령제3～12주적표본절편,매격10장추출1장절편,행면역조직화학염색.주요관찰지표:제3～12주배태간간세포갑태단백、c-Met、세포각단백19적표체.결과:제3～5주갑태단백、c-Met정양성반응,제시위간간세포;제10～12주갑태단백、c-Met양성세포주요분포우회관구주위,제시차시간간세포주요국한우회관구주위적간색내,여성년간중란원세포(성년간간세포)적분포일치.세포각단백19양성반응재제7주시개시출현;제10～11주시세포각단백19양성반응주요위우회관구부근적간색세포화담관판세포급담관상피세포;제12주시세포각단백19양성신호의견우담관판화담관상피세포.차시소유적담관판세포급담관상피세포균정갑태단백、c-Met화세포각단백19양성.결론:세포각단백19재간간세포중무표체,부재담관상피세포급기조세포중유표체,가능불괄합작위간간세포적표지물.소유적담관판급담관상피세포균정갑태단백、c-Met、세포각단백19양성반응,추측갑태단백+/c-Met+/세포각단백19+적세포가능위담관조세포.
BACKGROUND: There are few studies conceming morphological charactedstics, space-time distribution and differentiation of hepatic stem cells during embryo liver development.OBJECTIVE: To understand the action of alpha fetoprotein, cytokeratin-19 (CK19) and c-Met in the liver through observing the expression of them.DESIGN, TIME AND SETTING: The in vitro cytological observational study was performed at the Central Laboratory of Weifang Medical College from June 2005 to December 2006.MATERIALS: A total of 40 embryo samples were obtained from 3-rnonth aborted fetus, which were supplied by Hospital Affiliated to Weifang Medical College.METHODS: Aborted embryo was collected and made into sections within 30 minutes. Fetal age was defined according to embryonic layer formation, somite number and organ development under a microscope. Sample sections with fetal age of 3-12weeks were selected. One was collected from every eleven sections and underwent immunohistochemical staining.MAIN OUTCOME MEASURES: Expression of alpha fetoprotein, CK19 and c-Met was measured in embryo hepatic stem cells aged 3-12 weeks.RESULTS: At 3-5 weeks, samples were positive for alpha fetoprotein and c-Met, which were indicated as hepatic stem ceils. At 10-12 weeks, alpha fetoprotein- and c-Met-positive cells were mainly distributed surrounding the header, which suggested that hepatic stem cells were mainly located at hepatic cord of the header. This had similar distribution as adult hepatic oval cells (adult hepatic stem cells). CK19-positive reaction was found at week 7, and mainly at hepatic cord cells, bile duct sheet cells and bile duct epithelial cells at 10-11 weeks. CKlg-positive reaction was only seen at the bile duct sheet and bile duct epithelial cells at week 12. At this time, all bile duct sheet cells and bile duct epithelial cells were positive for alpha fetoprotein, c-Met and CK19.CONCLUSION: CK19-positive reaction was not found in hepatic stem cells, but only detected in bile duct epithelial cells and progenitor calls. CK19 may be not fit for a marker of hepatic stem cells. All bile duct sheet and bile duct epithelial cells are positive for alpha fetoprotein, c-Met and CK19. It is assumed that alpha fetoprotein+/c-Met+/CK19+ may be bile duct progenitor calls.