Chang I-C, Wu J-Y, Lu H-I, Ko H-W, Kuo J-L, Wang C-Y, Shen P-S, Hwang S-M
Bioresources Collection and Research Center, Food Industry Research and Development Institute, Hsinchu 30062, Taiwan, ROC.
Cell Prolif. 2006 Dec;39(6):457-69. doi: 10.1111/j.1365-2184.2006.00404.x.
Infection of freshly isolated and cryopreserved lymphocytes with Epstein-Barr virus (EBV) leads to the establishment of human B lymphoblastoid cell lines (LCLs). Techniques for optimal infection of the lymphocytes are vital for the establishment of a human biobank. The present study found that more than half (58-86%) of such established LCLs had transport times of less than 48 h, cell densities exceeding 10(6) cells/ml and cell viabilities greater than 90%. After EBV infection, 3306 freshly isolated lymphocytes required 30.0 +/- 0.1 days to become LCLs. Conversely, 1210 cryopreserved lymphocytes required 36.2 +/- 0.4 days. Cell density and viability of the culture affected transformation time in freshly isolated lymphocytes. On the other hand, blood transport time, cryopreservation time and initial cell viability were major factors in cryopreserved specimens. These results contribute to general information concerning the establishment of a human biobank for EBV infected cells.
用爱泼斯坦-巴尔病毒(EBV)感染新鲜分离和冷冻保存的淋巴细胞可建立人B淋巴母细胞系(LCLs)。淋巴细胞的最佳感染技术对于建立人类生物样本库至关重要。本研究发现,超过一半(58 - 86%)的此类已建立的LCLs运输时间少于48小时,细胞密度超过10⁶个细胞/毫升,细胞活力大于90%。EBV感染后,3306个新鲜分离的淋巴细胞需要30.0±0.1天成为LCLs。相反,1210个冷冻保存的淋巴细胞需要36.2±0.4天。培养物的细胞密度和活力影响新鲜分离淋巴细胞的转化时间。另一方面,血液运输时间、冷冻保存时间和初始细胞活力是冷冻保存样本中的主要因素。这些结果有助于提供有关建立EBV感染细胞人类生物样本库的一般信息。