Construction of Somatic Cell Hybrid Lines: Fusion of Mouse Thymidine Kinase-Deficient 3T3 Fibroblasts and Human Lymphoblastoid Cells

Somatic cell hybrids are generated by fusion of two different parental cells. This technology has been used extensively in the production of monoclonal antibodies and has made significant contributions to the field of human genetics through its applications in gene expression, gene mapping, and positional cloning of human disease genes. In our laboratory, we have employed this technique in the positional cloning of several genes for human diseases associated with cytogenetic abnormalities (chromosomal disorders), including translocations. Somatic cell hybrids are constructed by fusing mouse thymidine kinase-deficient 3T3 fibroblasts with human lymphoblastoid cells, as a result of which specific hybrid cells containing only cytogenetically abnormal human chromosomes involved in a chromosomal disorder can be successfully isolated and cloned. These hybrid cells serve as an excellent tool with which to define the exact chromosomal breakpoints involved in a cytogenetic abnormality and to identify genes at the breakpoints.