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犬肾细胞系的染色体组型分析与致癌/致瘤性的实验研究 被引量:4

Experimental Research on Carcinogenesis or Tumorigenicity of MDCK Canine Kidney Cell(CKC) Lines & Analysis of Their Chromosome Karyotypes
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摘要 以人类子宫颈癌细胞系Hela为阳性对照 ,以连传 3代的犬、猫肾原代细胞 (CKC、FKC)为阴性对照 ,对来自国内外不同单位收集的 4株MDCK传代细胞系培养 2 5~ 4 5代的完整活细胞或冻融裂解细胞进行裸鼠致癌 /致瘤实验观察 ,筛选出致癌性极低、符合细胞遗传学要求、无传染因子污染的几株MDCK细胞系用于制苗 ,并建立了相应的细胞种子库和工作库 ,供科研和生产使用 ,4年运转很好。不同代次MDCK细胞系染色体众数所占比率的相差率一般不超过 5 %~ 15 % ,结构畸变率一般为 0~ 3%。研究表明 ,MDCK细胞染色体遗传特征决定致瘤性质 ,并具有种属特异性 ,MDCK细胞不论核型如何 ,始终具有致癌性 ,但其致癌 /致瘤性差 ,只有超二倍体以上细胞才具有高的致癌 /致瘤率 (如YA株的为 2 8/ 5 8) ,亚二倍体细胞的致癌 /致瘤率很低 (其它 3株MDCK细胞的致癌 /致瘤率为 5 / 5 4 ) ,且一般致上皮源性恶性肿瘤 ,多为高中分化腺癌。冻融裂解癌细胞系 (X株Hela、JA株Vero、M株BHK 2 1、YA株MD CK)的致癌 /致瘤性相应降低 ,极低致癌 /致瘤性细胞系 (M株或JC株MDCK)不会因冻融裂解而增加致癌 /致瘤性。证明MDCK细胞系冻融裂解物不致癌 ,降低制苗毒液中细胞系基因含量 ,完全可以将MDCK细胞系 (M ,JB ,JC株 )用于犬五联苗生产。MDCK细胞? The chromosomal number variations and structural aberrations of the MDCK cell line, primary feline or canine kidney cell(FKC or CKC) and Hela cell line were investigated and their karyotypes of conventional chromosome bands were analyzed. The carcinogenesis or tumorigenicity testing of these cell lines in about 232 nude mice for colony formation in soft agarose and for haemagglutination under different concentration of plant lectins of these cells were carried out. It was the prerequisite that the incidence of cancer or tumor in nega tive control nude mice inoculated subcutaneously with primary feline or canine kidney cell cultures purified in vitro at passage 3 was 0 (0/22) and 0 (0/10), respectively. The incidence of the progressively growing malignant tumor(MT) in positive control nude mice inoculated subcutaneously with Hela cell cultures of KB, X, or NM20/X strain was 10/10, 25/25 and 5/5l, respectively. The results showed that the incidence of tumor in nude mice with tetraploid YA strain of MDCK cell during 20~45 passages, with hypodiploid JB strain of MDCK cell on passage 25, with di and hypoploid JC strain of MDCK cell during 2~15 passages or with hypoploid M strain of MDCK cell during 9~27 passages was 28/58, 1/5, 4/18 and 0/31, respectively. The chromosomal analysis results showed that the ratio of difference in the rate of modal chromosome numbers between high(mcs+n) and lowest (mcs)passages was not more than 5%~15% and the structure aberrations were generally 0~ 3%. These results proved that the genetic characteristics of chromosomal number of cell lines determines their tumorigenicity, but it is species specific. The MDCK line has tumorigenicity no matter what its chromosome karyotype is, at least it has very low tumorigenicity even when its modal chromosome number is hypoploid. The repeatedly frozen, thawed and split controls of tumorigenicity positive cell lines(X strain of Hela, M strain of BHK 21, JA strain of Vero, YA strain of MDCK) have much lower tumorigenicity or are even non carcinogenesis. And the repeatedly frozen, thawed and split controls of very low tumorigenicity cell lines (M or JC strain of MDCK) are certainly non carcinogenic and never have increased tumorigenicity. It is thus evident that MDCK cells of M, JB or JC strains can be approved as substrate for the preparation of attenuated viral vaccines, but the MDCK cells of YA strain can not be approved as substrate for the preparation of attenuated viral vaccines. In summary, all strains of MDCK cell line have at least low tumorigenicity, and never have non cancinogenic MDCK. Very low tumorigenicity MDCK cell strains can certainly be used for the production of canine viral vaccines if the DNA content in the viral cell cultures is significantly decreased through conventional means during the manufacturing process. Therefore, the master cell stock and working cell bank of the MDCK line used for vaccine manufacture were established in China. They are free of infectious agents, and described with respect to cytogenetic characteristics and tumorigenicity.Tests showed that there were correlations among cell line chromosome number variations, anchorage independence in soft agarose, haemagglutination under plant lectins, and tumor forming ability in nude mice. Thus all the in vitro tests were an economical, simple and reliable means for monitoring the tumor forming ability of the MDCK line in nude mice.
出处 《中国农业科学》 CAS CSCD 北大核心 2002年第5期562-570,T003,共10页 Scientia Agricultura Sinica
基金 国家火炬计划重大项目
关键词 疫苗生产 犬肾细胞系 染色体组型 致癌 致瘤性 实验研究 Cell line MDCK Hela Feline or canine kidney cell Karyotypes Carcinogenesis or tumorigenicity
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