Messer Peters P G, Kamarck M E, Ruddle F H
J Immunol Methods. 1983 Aug 12;62(1):39-47. doi: 10.1016/0022-1759(83)90108-4.
Genetic mapping of differentiation specific surface antigens has been hampered by difficulty in preparation of interspecies hybrid cells which continue to express differentiated functions. A method has been developed for production of interspecies T cell hybrids which continue to express T cell specific cell surface molecules. Hybrids were constructed from either the human leukemic cell line MOLT-4 or freshly isolated human peripheral blood T cells and the mouse T lymphoma line BW5147. Optimal fusion efficiency resulted with pre-treatment of the human parental line with phytohemagglutinin followed by hybridization with 40% polyethylene glycol and plating without thymocyte feeder layers. Immortalization of hybrid lines was accomplished through addition of rat T cell growth factor to cultures.
分化特异性表面抗原的遗传图谱绘制一直受到阻碍,原因是难以制备能持续表达分化功能的种间杂交细胞。现已开发出一种方法来生产能持续表达T细胞特异性细胞表面分子的种间T细胞杂交体。杂交体由人白血病细胞系MOLT-4或新鲜分离的人外周血T细胞与小鼠T淋巴瘤系BW5147构建而成。通过用植物血凝素预处理人亲本细胞系,然后与40%聚乙二醇杂交并在无胸腺细胞饲养层的情况下铺板,可获得最佳融合效率。通过向培养物中添加大鼠T细胞生长因子来实现杂交细胞系的永生化。