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目的改进EB病毒转化冻存全血为人B淋巴母细胞的方法,提高转化效率,克服远距离分散采样造成的实验不便,建立新疆和田维吾尔族长寿老人永生细胞株。方法采集新鲜肝素抗凝血样,冻存时添加小牛血清和DMSO作为冷冻保护液,复苏后以新鲜制备的EB病毒进行转化,对照组为仅添加10%DMSO的传统冻存全血法。结果以改进的冻存全血法成功建立了9株新疆和田维吾尔族长寿老人永生细胞,建系成功率为100%(9/9),以传统冻存全血法建系成功一例(1/14)。HLA基因分型结果在建系前后完全一致。结论与传统冻存全血法相比,改进后的方法能更有效地转化长寿老人永生细胞株,可为长寿的分子遗传机制研究以及某些细胞生物学性状的检测提供充足的实验材料。
Objective To improve the method of transforming whole blood of frozen EB virus into human B lymphoblastoid cells and improve the transformation efficiency to overcome the inconvenience caused by long distance scattered sampling and establish the immortal cell line of longevity elderly in Wada of Xinjiang. Methods Fresh heparin anticoagulant blood samples were collected. During cryopreservation, bovine serum and DMSO were added as cryoprotectants. After resuscitation, freshly prepared EB virus was used for transformation. The control group was a traditional cryopreserved whole blood method with only adding 10% DMSO. Results Nine immortalized human longevity human beings were successfully established in the frozen whole blood of Hetian with a success rate of 100% (9/9). The traditional cryogenic whole blood method was successfully established in one case (1/14 ). HLA genotyping results were exactly the same before and after the establishment of the department. Conclusion Compared with the traditional cryopreserved whole blood method, the improved method can transform the immortalized cell line of longevity elderly more effectively, which can provide sufficient experimental materials for molecular mechanism study of longevity and the detection of some cell biological traits.