CAJ | 학술논문

目的：建立新的胶质瘤细胞株,为肿瘤研究提供具备新特性的细胞工具。方法临床手术来源的胶质瘤组织进行培养传代,第10代后克隆建立细胞株( SHG139)。经免疫荧光细胞化学技术检测蛋白表达,流式细胞术( FCM)分析细胞增殖和细胞周期,染色体核型分析SHG139细胞的生物学特性。应用神经干细胞培养液( NSCM )培养 SHG139细胞,使其转化成胶质瘤干细胞球(SHG139s),并检测SHG139s细胞的干性标志物。诱导分化SHG139s细胞后,检测分化细胞的胶质前体神经节苷脂(A2B5)、胶质纤维酸性蛋白(GFAP)、β-Ⅲ tubulin和半乳糖神经酰胺(GalC)的表达情况,进一步比较SHG139和SHG139s细胞颅内原位移植瘤的病理特征。结果成功建立的SHG139第20代和第60代细胞A2B5、GalC、GFAP、酸性钙结合蛋白(S-100)和vimentin表达均为阳性,与肿瘤组织的免疫组化结果及病理特征一致。 SHG139细胞传代后24 h内增殖显著,染色体总数为68条,多倍体较多。 SHG139s细胞中A2B5、巢蛋白(nestin)和神经元-神经胶质2型抗原(NG2)阳性比例分别为(84.12±9.96)%、(73.86±5.01)%和(73.37±2.09)%。 SHG139s细胞诱导分化后,贴壁细胞中GFAP、β-Ⅲ tubulin 和 GalC 的阳性比例分别为(92.89±2.24)%、(64.85±4.09)%和(33.57±4.14)%。 SHG139s细胞颅内移植瘤呈胶质肉瘤和星形-少枝胶质细胞瘤的多种形态,肿瘤血管和瘤周血管以鼠源性为主。结论 SHG139为星形胶质瘤细胞株, NSCM 培养可成功获得 SHG139s。SHG139s为A2B5+ CD133-亚群细胞,具有自我更新和多向分化潜能。与SHG139细胞颅内移植瘤比较,SHG139s细胞颅内移植瘤具有更强的恶性度和侵袭性。
목적：건립신적효질류세포주,위종류연구제공구비신특성적세포공구。방법림상수술래원적효질류조직진행배양전대,제10대후극륭건립세포주( SHG139)。경면역형광세포화학기술검측단백표체,류식세포술( FCM)분석세포증식화세포주기,염색체핵형분석SHG139세포적생물학특성。응용신경간세포배양액( NSCM )배양 SHG139세포,사기전화성효질류간세포구(SHG139s),병검측SHG139s세포적간성표지물。유도분화SHG139s세포후,검측분화세포적효질전체신경절감지(A2B5)、효질섬유산성단백(GFAP)、β-Ⅲ tubulin화반유당신경선알(GalC)적표체정황,진일보비교SHG139화SHG139s세포로내원위이식류적병리특정。결과성공건립적SHG139제20대화제60대세포A2B5、GalC、GFAP、산성개결합단백(S-100)화vimentin표체균위양성,여종류조직적면역조화결과급병리특정일치。 SHG139세포전대후24 h내증식현저,염색체총수위68조,다배체교다。 SHG139s세포중A2B5、소단백(nestin)화신경원-신경효질2형항원(NG2)양성비례분별위(84.12±9.96)%、(73.86±5.01)%화(73.37±2.09)%。 SHG139s세포유도분화후,첩벽세포중GFAP、β-Ⅲ tubulin 화 GalC 적양성비례분별위(92.89±2.24)%、(64.85±4.09)%화(33.57±4.14)%。 SHG139s세포로내이식류정효질육류화성형-소지효질세포류적다충형태,종류혈관화류주혈관이서원성위주。결론 SHG139위성형효질류세포주, NSCM 배양가성공획득 SHG139s。SHG139s위A2B5+ CD133-아군세포,구유자아경신화다향분화잠능。여SHG139세포로내이식류비교,SHG139s세포로내이식류구유경강적악성도화침습성。
Objective To establish a new glioma cell line and analyze its biological characteristics, and to provide a useful cellular tool with new features for cancer research. Methods Glioma tissue was taken from surgical specimen clinical of a clinical patient. Primary culture was carried out, and a cell line ( SHG139 ) was established after 10 passages. Immunofluorescence staining was performed to detect the expression of proteins, and cell proliferation and cycle were detected by flow cytometry method ( FCM) . The biological characteristics of SHG139 cells were detected by chromosome karyotype analysis. SHG139s glioma cells derived from SHG139 glioma cell line were cultured with neural stem cell medium. Then stem cell markers were determined. SHG139s cells were induced with serum-containing medium, and their expression of A2B5, GFAP,β-Ⅲtubulin, and GalC was detected. Intracranial xenograft tumor of both SHG139 glioma cells and SHG139s glioma stem cell spheres was generated in rats. Results The expressions of A2B5, GalC, GFAP, S-100, and vimentin in the 20 and 60 passages of SHG139 cells were positive, consistent with the immunohistochemical results and pathological features. SHG139 cells proliferated significantly within 24 h after subculture, and their total number of chromosomes was 68 and mostly multiploid. They were positive for A2B5 (84. 12 ± 9. 96)%, nestin (73. 86 ± 5. 01)%, and NG2 (73. 37 ± 2. 09)%. SHG139s cells were induced, and the ratio of positive cells of GFAP, β-Ⅲ tubulin and GalC was (92. 89 ± 2. 24)%,(64. 85 ± 4. 09)% and (33. 57 ± 4. 14)%, respectively. Conclusions SHG139 is an astroglioma cell line, from which SHG139s cells can be successfully obtained by culture with NSCM. SHG139s cells are of A2B5+/CD133-GSCs subgroup cells, with potentials of self-renewal and multi-directional differentiation. Compared with the intracranial SHG139 xenograft tumor, the intracranial SHG139s xenograft tumor is more malignant and aggressive.