兔脾脏成纤维型细胞RS-17系的建立及其生物学特性
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摘要
采用胰酶消化法培养技术,对乳兔脾脏组织的细胞进行原代培养,从脾脏组织中分离出1株成纤维型细胞,命名为RS-17。通过对细胞传代培养条件优化,细胞标志性蛋白检测,细胞生长曲线分析,细胞核型以及细胞致瘤性等测定,结果显示,RS-17细胞具有血清依懒性,成纤维细胞标志性蛋白检测阳性,具有正常的核型,无致瘤性,且目前细胞传至95代仍可稳定生长。这为RHDV(兔出血症病毒)感染机制等相关研究提供了细胞材料。
The cells of the spleen tissue of the rabbit were primary cultured by trypsin digestion technique,and a fibroblast-like cell was isolated from the spleen tissue and named as RS-17. The subculture conditions was optimized,cellular marker proteins were detected by immunofluorescence,and the growth curve was analyzed,karyotype and tumorigenicity of the RS-17 were detected. The results showed that the RS-17 was fibroblast cell and had the normal karyotype,normal growth characteristic,no tumorigenicity. The cell has been passaged stably to the 95 generations. These results provide cellular material for studies related to the infection mechanism of rabbit hemorrhagic disease virus( RHDV).
The cells of the spleen tissue of the rabbit were primary cultured by trypsin digestion technique,and a fibroblast-like cell was isolated from the spleen tissue and named as RS-17. The subculture conditions was optimized,cellular marker proteins were detected by immunofluorescence,and the growth curve was analyzed,karyotype and tumorigenicity of the RS-17 were detected. The results showed that the RS-17 was fibroblast cell and had the normal karyotype,normal growth characteristic,no tumorigenicity. The cell has been passaged stably to the 95 generations. These results provide cellular material for studies related to the infection mechanism of rabbit hemorrhagic disease virus( RHDV).
引文
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[13]魏如雪,赵学明,郝海生,等.兔胎儿成纤维细胞的分离培养和鉴定[J].畜牧兽医学报,2016,47(6):1272-1279.
[2] HONDA A,HIROSE M,HATORI M,et al. Generation of in-duced pluripotent stem cells in rabbits:potential experimentalmodels for human regenerative medicine.[J]. Journal of BiologicalChemistry,2010,285(41):31362.
[3]詹康,左晓昕,陈银银,等.猪小肠上皮细胞分离培养与鉴定[J].动物营养学报,2015,27(5):1477-1484.
[4]刘学强.转染h TERT基因建立人永生化神经干细胞系的实验研究[D].重庆:第三军医大学,2005.
[5]高雯,臧荣余,王雁,等.人类输卵管上皮永生化细胞系的建立及鉴定[J].中国癌症杂志,2013,23(4):241-247.
[6]袁文斌.人源性成纤维细胞系的建立及其生物学特性初步研究[D].杭州:浙江大学,2004.
[7] SMITH J L,LEE L C,READ A. One-step immortalization of pri-mary human airway epithelial cells capable of oncogenic transfor-mation[J]. Cell Biosci,2016,6:57.
[8] VEITONMAKI N,FUXE J,HULTDIN M. Immortalization of bo-vine capillary endothelial cells by hTERT alone involves inactiva-tion of endogenous p16INK4A/pRb[J]. FASEB J,2003,17(6):764-766.
[9]洪海霞.永生化猪脐静脉血管内皮细胞系的建立及其生物学特征分析[D].西安:西北农林科技大学,2007.
[10]苏正元,张彦明,洪海霞,等.应用端粒酶逆转录酶基因使猪血管内皮细胞永生化的初步研究[J].畜牧兽医学报,2007,38(4):407-411.
[11]吉传义,杜念兴,徐为燕,等.兔出血症病毒细胞培养的初步研究[J].病毒学报,1992,8(8):252-256,293.
[12]莫国艳,田代志,张丽萍,等.兔Tenon’s囊组织原代成纤维细胞分离培养与鉴定[J].武汉大学学报(医学版),2017,38(6):973-977.
[13]魏如雪,赵学明,郝海生,等.兔胎儿成纤维细胞的分离培养和鉴定[J].畜牧兽医学报,2016,47(6):1272-1279.