西伯利亚狍成纤维细胞建系及其生物学特性分析北大核心CSCD

本研究以内蒙古大青山获得野生雄性和雌性西伯利亚狍(Capreolus pygargus)为实验材料,利用组织块贴壁培养法进行气管、肺和耳3种组织成纤维细胞原代建系,研究不同组织来源的细胞贴壁率、冷冻前及复苏后存活率、生长曲线,进一步绘制狍成纤维细胞核型图并分析其G带特征。实验结果显示,气管、肺和耳3种组织成纤维细胞增殖经历潜伏期、对数生长期、平台期三个阶段,细胞形态为梭形、三角形或不规则形,是典型成纤维细胞形态;成纤维细胞呈漩涡状生长,其中气管、耳成纤维细胞生长增殖能力最强、肺成纤维细胞增殖能力较弱,气管和耳组织来源成纤维细胞呈典型“S”型细胞生长特征。染色体核型及G带分析结果显示,雄性狍成纤维细胞染色体条数为2n=70,其中,有34对常染色体,形态类型为12条近端着丝粒染色体(st),22条亚中着丝粒染色体(sm),1对性染色体,X染色体为中着丝粒染色体(m),Y染色体为近端着丝粒染色体(st),5条超数染色体(B);雌性狍成纤维细胞染色体条数为2n=70,其中,有34对常染色体,其形态类型为29条亚中着丝粒染色体(sm),5条近端着丝粒染色体(st),1对为性染色体,X染色体为亚中着丝粒染色体(sm),8条超数染色体(B)。本研究成功建立了雄性和雌性西伯利亚狍气管、肺和耳3种组织来源的成纤维细胞系,在体外培养时生长状态良好且维持了细胞的遗传信息稳定性,绘制了西伯利亚狍雄性和雌性染色体核型及G带图谱,为将来更深入开展相关研究提供材料与基本技术支撑。

In Vitro Culture of Fibroblasts and Analysis of Their Biological Characteristics in Siberian Roe Deer

Methods]In this study, ear, lung and trachea samples of male and female Roe Deer were collected from Da Qing Mountains in Inner Mongolia Autonomous Region. The primary fibroblast lines of trachea, lung and ear were established by tissue adherence method. Adherent rate, survival rate and growth curve of cells from different tissues were explored before freezing and after resuscitation. The karyotype of Roe Deer fibroblasts and G-band characteristics were performed in this study.Results]The results showed that the proliferation of fibroblasts in trachea, lung and ear experienced three stages: incubation period, logarithmic growth period and plateau period. The cell morphology was spindle, triangular or irregular, which was a typical fibroblast morphology; Fibroblasts grew in a vortex, in which the growth and proliferation ability of tracheal and ear fibroblasts was the strongest, and the proliferation ability of lung fibroblasts was slow. Fibroblasts derived from tracheal and ear tissues showed typical "S" growth characteristics. The results of karyotype and G-band analysis showed that the number of chromosomes of fibroblasts was 2n = 70 in male Roe Deer, BS = 5, including 34 pairs of autosomes, the morphological type was 12st + 22sm, and one pair was sex chromosome, the morphological type X was m, Y was St; The number of chromosomes of fibroblasts is 2n = 70 in female roe deer, BS = 8, including 34 pairs of autosomes, the morphological type is 29sm + 5st, one pair is sex chromosome, and the morphological type X is SM. Conclusion]In conclusion, fibroblast lines derived three tissue were successfully established in male and female Roe Deer. Genetic information of cells were stable in vitro. The karyotype and G-band analysis of male and female Roe Deer were firstly investigated. This study provided theoretical basic technical guidance for related research in the future.