The generation of Ig-secreting UC 729-6 derived human hybridomas by electrofusion

Abstract

UC 729-6, a 6-thioguanine resistant human lymphoblastoid B cell line, was fused with human lymphocytes by electrofusion. Resulting human-human hybridomas were tetraploid, expressed markers derived from both fusion parents, and secreted approximately 1 microgram Ig/10(6) cells/ml/day. Cells to be used for electrofusion were washed in 0.3M mannitol, and fusions were performed with glass slides, 1.0 ml, and 50.0 ml chambers. Fusion sequences consisted of alignment, compression, and the fusion event itself. The optimal cell concentration for electrofusion was 5 X 10(6)/ml. Fusion efficiencies of human lymphocytes were ranked as follows: lymphoma cells greater than lymph node lymphocytes greater than PBL. 80% of the lymphoma hybridomas, 60% of the lymph node hybridomas, and 40% of the PBL hybridomas were Ig secretors. These data demonstrate that the UC 729-6 cell line is a suitable vector for generating human-human hybridomas by electrofusion.