CAJ | 학술논문

目的:从切除的人小肠间质瘤(gastrointestinal stromal tumors,GIST)组织建立一株间质瘤细胞系,初步鉴定其生物学特性.方法:取人小肠间质瘤组织切成小块,置于含10%胎牛血清、青霉素、链霉素和庆大霉素的RPMI 1640培养基中进行培养,待细胞长满90%汇合面后消化传代培养,并进行形态学、生长动力学、免疫组织化学(免疫组化)及致瘤性等生物学特性研究.结果:成功建立GIST细胞系,命名为GIST-H1 细胞系.该细胞系已在体外培养生存1年,传60余代.免疫组织化学分析显示该细胞CD117(+),SMA(+),dog-1(+),CD34(-)及S-100(-);该细胞系体外生长的倍增时间为47.5 h;软琼脂集落形成率为24.8%;透射电镜下可观察到该细胞富含线粒体和内质网,多数细胞表面无绒毛,胞核呈卵圆形;染色体核型分析为非整倍体,众数为60～98条;该细胞接种至裸鼠皮下后致瘤性为100%.结论:间质瘤细胞系在体外长期培养后已永生化,形成了可连续传代的细胞系,且具有明显的间质瘤细胞恶性表型特征.
목적:종절제적인소장간질류(gastrointestinal stromal tumors,GIST)조직건립일주간질류세포계,초보감정기생물학특성.방법:취인소장간질류조직절성소괴,치우함10%태우혈청、청매소、련매소화경대매소적RPMI 1640배양기중진행배양,대세포장만90%회합면후소화전대배양,병진행형태학、생장동역학、면역조직화학(면역조화)급치류성등생물학특성연구.결과:성공건립GIST세포계,명명위GIST-H1 세포계.해세포계이재체외배양생존1년,전60여대.면역조직화학분석현시해세포CD117(+),SMA(+),dog-1(+),CD34(-)급S-100(-);해세포계체외생장적배증시간위47.5 h;연경지집락형성솔위24.8%;투사전경하가관찰도해세포부함선립체화내질망,다수세포표면무융모,포핵정란원형;염색체핵형분석위비정배체,음수위60～98조;해세포접충지라서피하후치류성위100%.결론:간질류세포계재체외장기배양후이영생화,형성료가련속전대적세포계,차구유명현적간질류세포악성표형특정.
Objective To establish the cell line from specimens of resectable human gastrointestinal stromal tumors (GIST) and to verify the characteristics of cell biology in vitro. Methods The tissues from biopsies of human GIST were cultured in RPMI 1640 media supplemented with 10% fetal bovine serum. After growing to 90% confluence,the cells were detached for subculture and their characteristics,including morphology,growth kinetics,karyotype analysis,immunohistochemical analysis and tumorigenicity in nude mice were determined. Results GIST named GIST-H1 was successfully established. The cell line was passaged for more than 60 times 1 year. The characteristics demonstrated: The population doubling time calculated in the log phase of growth was 47.5 h. The cloning efficiency in the soft agar averaged 24.8%.Electronmicroscopically,there were rich ribosomes and mitochondrion in the cytoplasm. Immunohistochemical analysis showed CD117(+),SMA(+),dog-1(+),CD34(-),and S-100(-).Karyotype analysis illustrated aneuploidy with the modal chromosomal number 60-98.The GIST cells transplanted in nude mice had high tumorigenicity. Conclusion The immortalized GIST cells are devoloped in vitro and have specific characteristics of GIST.