应用微量冻存法建立EB病毒转化脐带血淋巴母细胞系

发布时间：2018-12-16 17:44

【摘要】：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。
[Abstract]:Objective to establish the transformation of EB virus (EBV) into umbilical cord hemolymph mother cell line (LCLs), by microcryopreservation to provide a basis for the permanent preservation of cord blood resources with genetic diseases. Methods Cord blood samples were collected from 20 pregnant women with informed consent from the middle to the third trimester of pregnancy. The chromosome G-banding analysis was performed after LCLs, cryopreservation and resuscitation of cord blood were obtained by EBV transformation. The karyotype stability before and after the establishment of the system was tested. Results among the 20 cases, 19 cases were successfully transformed by microcryopreservation, the success rate was 95%, and the successful rate of resuscitation of umbilical cord blood LCLs after cryopreservation was 100%. Except for one case of fetal failure due to fetal edema, 19 cord blood samples could be observed to be mother cell enlarged and proliferating lymphoblastoid cells condensed into clusters at about 10 days. After 6 to 8 weeks, stable cord blood LCLs, could be obtained. There was no significant difference in karyotype of lymphocyte before and after transformation. Conclusion the cytogenetic characteristics of cord blood LCLs transformed by umbilical cord blood LCLs,EB virus can be successfully established by microcryopreservation, which can be used for further genetic research.
【作者单位】：广东省妇幼保健院医学遗传中心;
【基金】：广东省医学科研基金立项课题(编号:B2014024)
【分类号】：R329.2

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