Generation of sheep X (sheep X mouse) heterohybridoma cell line expressing the beta-1 integrin membrane molecule

Sheep are an important biological model in such diverse areas as immunology and reproductive biology. The limitation of sheep as an experimental model is the absence of reliable cell lines. To establish cell lines that express functional sheep membrane molecules, we produced a sheep x mouse heterohybridoma by fusion of sheep efferent lymph T cells with the murine myeloma cell line NS1. A cloned heterohybridoma fusion partner was selected by treatment with 8-azaguanine. The resulting cell line HL1/385 was selected for hypoxanthine/aminopterin/thymidine (HAT) sensitivity and growth efficiency. The HL1/385 cell line was used as a back-fusion partner into lectin-stimulated efferent T lymphocytes. The back-fusion approach produced more than 50 heterohybrid cell lines with high growth efficiency. The expression of physiological levels of the sheep beta-1 integrin cell surface molecule on the HT4/6 cell line was stable for months in culture. These results suggest that somatic heterohybrids may provide a reliable source of cell lines for sheep studies in vitro.