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The electronic form of this document may be cited in the following style:
Human Genome Program, U.S. Department of Energy, DOE Human Genome Program Contractor-Grantee Workshop IV, 1994.
Abstracts scanned from text submitted for November 1994 DOE Human Genome Program Contractor-Grantee Workshop. Inaccuracies have not been corrected.
A Panel Of Mouse/Human Monochromosomal Hybrid Cell Lines Each Containing A Single Different Tagged Human Chromosome
Arbansjit K. Sandhu, G. Pal Kaur and Raghbir S. Athwal
Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine 3420 N. Broad Street, Philadelphia, PA 19140
We are producing a panel of mouse/human monochromosomal hybrid cell lines each containing a single different gpt tagged human chromosome. Presence of the selectable marker on the human chromosome assures stable retention while cells are cultured in the medium containing mycophenolic acid (25ug/ml) and xanthine (70ug/ml, MX medium). This panel would provide a unique resource for gene mapping and gene isolation. In addition tagged human chromosomes can be selectively introduced into any cell type of interest for genetic analysis of complex phenotypes by complementation.
The experimental approach to produce these cell lines involves tagging of the chromosomes in normal diploid human cells with a retroviral vector. Since normal human cells have a limited life span, they are fused with mouse A9 cells to perpetuate the marked chromosomes. Tagged human chromosomes which are now present in mouse/human hybrid cells are transferred further to mouse and/or Chinese hamster cells by microcell fusion method. Monochromosomal hybrid cell lines thus recovered are analyzed by a battery of methods including G-11, chromosome painting and Alu-PCR Southern hybridization, to ascertain the identity and integrity of the transferred human chromosome. We have already produced and characterized monochromosomal hybrid cell lines for 19 different chromosomes. This partial panel is comprised of hybrid cell lines for chromosomes 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10, 11, 12, 13, 14, 16, 17, 19, 21 and X. In addition to the hybrids containing intact chromosomes, cell lines bearing chromosomal arms have also been recovered and characterized.