Objective To establish EBV (Epstein-Barr virus, EBV) immortalized lymphoblastoid cell lines (lymphoblastoid cell line, LCLs) from small volumes of cryo-preserved umbilical cord blood, thus to provide the basis for permanent preservation of umbilical cord blood resources for genetic diseases.Methods Twenty umbilical cord blood samples were collected from women with pregnancy termination at the second or third trimester after informed consent.Immortalized LCLs from small volumes of cryo-preserved umbilical cord blood were established with EBV.After being frozen and recovered, the LCLs were detected by comparing chromosome G banding karyotype with the cell lines before immortalized.Results Nineteen LCL samples were successfully established, the success rate was 95%.The recovery rate of cell lines after being frozen was 100%.One case was failed for fetal hydrops.In the successful cases, large and clusters cells were observed on the 10th day, and developed into LCLs at the 6th-8th week.The karyotypes presented no significant difference before and after the immortalization.Conclusion The stable umbilical cord blood LCLs could be established after immortalized by EBV.%目的 应用微量冻存法建立EB病毒(EBV)转化脐带血淋巴母细胞系(LCLs),为永久保存具遗传疾病的脐带血资源提供研究基础.方法 知情同意下收集妊娠中期至晚期终止妊娠的20例孕妇脐带血样本,采用微量冻存法,通过EBV转化获得脐带血LCLs,冻存复苏后,染色体G显带分析,检测建系前后的核型稳定性.结果 20例样本中19例成功进行微量冻存法转化,成功率95%,转化成功的脐带血LCLs冻存后复苏成功率达100%.除1例因胎儿为水肿胎失败外,19例脐带血样本均可在10 d左右观察到淋巴细胞增大呈母细胞化,增殖的淋巴母细胞凝聚成团,6~8周后可获得稳定生长的脐带血LCLs,转化前后的淋巴细胞染色体核型无明显差异.结论 应用微量冻存法可成功建立脐带血LCLs,EB病毒转化的脐带血LCLs细胞遗传学特性稳定,可用于进一步的遗传学研究.
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