Molecular analysis of HPRT1+ somatic cell hybrids derived from a carrier of anHPRT1 mutation responsible for Lesch-Nyhan syndrome

Abstract

Heterozygous carriers of HPRT1 mutations responsible for Lesch-Nyhan syndrome can be detected by analysis of somatic cell hybrids derived from peripheral blood lymphocytes and Hprt1-negative cells of rodent origin followed by selection in culture medium containing hypoxanthine, aminopterine, and thymidine (HAT). The parental origin of the X chromosome containing the normal HPRT1 allele in HPRT1(+) hybrid cell lines can be determined by molecular haplotyping attributable to highly polymorphic X-linked markers. We used this procedure to study a presumed carrier whose paternal active X chromosome always segregated in the cell hybrids derived from her. Conversely, her maternal X chromosome was systematically absent in most cell hybrids, or when present, it was inactive and coexisted with an active, paternal X chromosome. These results clearly demonstrated that the proband was a heterozygous carrier of a mutation responsible for HPRT1 deficiency.

Akodon cursor(Rodentia: Cricetidae)

“…This capability is important in the identification of human HPRTl mutation responsible for Lesch-Nyhan syndrome (Rivero et al 2001 …”

Detailed analysis of X chromosome inactivation in a 49,XXXXX pentasomy

“…A cell hybrid panel was prepared with AKO1/15 cells derived from Akodon cursor [ 46 ]. Following a seven day growth in DMEM/6MP selective medium (Dulbecco's Minimal Essential Medium supplemented with 10% FCS and 6-mercaptopurine 16.7 μg/ml), 3 × 10 6 recipient cells were fused with 1.2 × 10 7 human lymphocytes [ 47 ]. Cell hybrids were selected in DMEM/HAT medium [DMEM with 10% FCS, hypoxanthine (100 μM), aminopterin (0.4 μM), thymidine (16 μM)].…”