Transfer of the human X chromosome to human-Chinese hamster cell hybrids via isolated HeLa metaphase chromosomes

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Abstract
Evidence is presented for the uptake of the human X chromosome by human-Chinese hamster cell hybrids which lack HPRT activity, following incubation with isolated human HeLa S3 chromosomes. Sixteen independent clonal cell lines were isolated in HAT medium, all of which contained a human X chromosome as determined by trypsin-Giemsa staining. The frequency of HAT-resistant clones was 32 × 10−6 when 107 cells were incubated with 108 HeLa chromosomes. Potential reversion of the hybrid cells in HAT medium was less than 5 × 10−7 The 16 isolated cell lines all contained activity of the human X-linked marker enzymes HPRT, PGK, α-Gal A, and G6PD, as determined by electrophoresis. The phenotype of G6PD was G6PD A, corresponding to G6PD A in HeLa cells. The human parental cells used in the fusion to form the hybrids had the G6PD B phenotype. The recipient cells gave no evidence of containing human X chromosomes. These results indicate that incorporation and expression of HeLa X chromosomes is accomplished in human-Chinese hamster hybrids which lack a human X chromosome.