Mevissen M L, Kwekkeboom J, Goormachtig E, Lindhout E, de Groot C
Laboratory of Cell Biology and Histology, University of Amsterdam, Netherlands.
Hum Antibodies Hybridomas. 1993 Apr;4(2):66-73.
Exposure of human B cells to Epstein-Barr virus (EBV) usually results in low frequencies of transformed cells. The transformed cells can be cloned poorly by limiting dilution, even when feeder cells are used. In recent years it has become clear that growth and antibody production of EBV-transformed cells are influenced by auto- and paracrine growth factors. Therefore, supernatants from the lymphoblastoid B cell lines JY and Raji were used as a source of growth factors to investigate their effect during EBV transformation of human B cells and consequently on cloning by limiting dilution of these transformed cells. Initial experiments to clone three established EBV-transformed B cell lines showed a strong increase in outgrowth of the number of cells in the presence of the supernatant (range: 1 per 2-8 of the originally plated cells) as compared to cells cultured without the supernatant (range: 1 per 17-100 of the originally plated cells). Transformation efficiencies of freshly isolated tonsil B cells were not influenced by the supernatant and were generally less than 1%. In contrast, transformation efficiency was increased up to 9.4% if B cells were both transformed and cultured in the presence of the supernatant. Cloning efficiencies increased if the cells used were transformed in the presence of the supernatant. Best results were seen when the supernatant was present during transformation and cloning of the cells. The presence of the supernatant during transformation and/or cloning of the B cells dramatically enhanced the number of B cells secreting IgG. Cloning of two established tonsil B cell lines resulted in a large number of B cell clones.(ABSTRACT TRUNCATED AT 250 WORDS)
人类B细胞暴露于爱泼斯坦-巴尔病毒（EBV）通常会导致转化细胞的频率较低。即使使用饲养细胞，通过有限稀释法也很难克隆转化细胞。近年来，越来越清楚的是，EBV转化细胞的生长和抗体产生受到自分泌和旁分泌生长因子的影响。因此，将淋巴母细胞B细胞系JY和Raji的上清液用作生长因子来源，以研究它们在人类B细胞EBV转化过程中的作用，以及由此对这些转化细胞进行有限稀释克隆的影响。对三种已建立的EBV转化B细胞系进行克隆的初步实验表明，与未添加上清液培养的细胞相比（范围：每17-100个原始接种细胞中有1个生长），在上清液存在的情况下，细胞生长数量显著增加（范围：每2-8个原始接种细胞中有1个生长）。新鲜分离的扁桃体B细胞的转化效率不受上清液影响，通常低于1%。相比之下，如果B细胞在有上清液的情况下进行转化和培养，转化效率可提高至9.4%。如果使用在上清液存在的情况下转化的细胞，克隆效率会提高。当细胞在转化和克隆过程中都有上清液时，效果最佳。B细胞在转化和/或克隆过程中上清液的存在显著增加了分泌IgG的B细胞数量。对两种已建立的扁桃体B细胞系进行克隆产生了大量B细胞克隆。（摘要截短于250字）
