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目的保存家族性疾病家系成员特有的基因组资源,为进一步研究奠定物质基础。方法应用 EB 病毒(Epstein-Barr Vires)转化外周血 B 淋巴细胞建立永生细胞系,分别采用4种方法:(1)外周血淋巴细胞转化法;(2)外周血淋巴细胞冻存转化法;(3)微量全血转化法;(4)微量全血冻存转化法。结果4种方法建系结果分别为:外周血淋巴细胞转化培养20株;外周血淋巴细胞冻存转化培养4株;微量全血转化培养4株;全血冻存转化培养4株,总计32株建系均获成功。成功转化的细胞体积增大,形态多样,增生的细胞多聚集成团。所有的细胞株冻存后复苏的成功率为100%。结论该研究提供了家族性疾病永生细胞系的建立方法,同时也为在分子水平上揭示家族性疾病的发病机理提供了 DNA 材料。
Objective To preserve the genomic resources unique to family members of familial diseases and lay the material foundation for further research. Methods The immortalized cell lines were established by transfection of peripheral blood B lymphocytes with Epstein-Barr Vires. Four methods were used respectively: (1) peripheral blood lymphocyte transformation; (2) peripheral blood lymphocyte cryopreservation; ( 3) trace whole blood conversion method; (4) trace whole blood cryopreservation method. Results The results of the four methods were as follows: 20 strains were transformed by peripheral blood lymphocytes; 4 were transformed by cryopreservation of peripheral blood lymphocytes; 4 were transformed by micro-whole blood; 4 were transformed by cryopreservation, 32 were total strains Department of construction have been successful. Successful transformation of the cells increased in size and diversity, proliferation of cells gathered in clusters. The success rate of all cell lines resuscitated after cryopreservation was 100%. Conclusions This study provides a method to establish an immortal cell lineage of familial disease and also provides a DNA material for revealing the pathogenesis of familial disease at the molecular level.