Тип публикации: статья из журнала
Год издания: 1998
Идентификатор DOI: 10.1002/(SICI)1098-2795(199806)50:2<128::AID-MRD2>3.0.CO;2-M
Ключевые слова: Chimeras, Hybrid cells, Mouse, Pluripotency, hypoxanthine phosphoribosyltransferase, animal cell, article, blastomere, cell differentiation, cell fusion, cell growth, cell synchronization, chimera, embryo cell, embryo development, germinal center, hybrid cell, nonhuman, priority journal, spleen cell, stem cell, X chromosome aberration, Alkaline Phosphatase, Animals, Biological Markers, Cell Line, Extracellular Matrix Proteins, Female, Intermediate Filament Proteins, Male, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Inbred CBA, Spleen, Stem Cells, X Chromosome, Animalia, Murinae, Rodentia, Vertebrata
Аннотация: Hypoxanthine phosphoribosyl-transferase-deficient (HPRT) mouse embryonic stem (ES) cells, HM-1 cells (genotype XY), were fused with adult female DD/c mouse spleen cells. As a result, a set of HAT-resistant clones was isolated. Four hybrid clones most similar in morphology and growth characteristics to the HM-1 cells were studied inПоказать полностьюdetail with respect to their pluripotency. Of these, three clones contained 41-43 chromosomes, and one clone was nearly tetraploid. All the clones had the XXY set of sex chromosomes and expressed the HPRT of the somatic partner only. The hybrid clones shared features with the HM-1 cells, indicating that they retained their pluripotent properties: (1) embryonic ECMA-7 antigen, not TROMA-1 antigen, was present in most cells; (2) the hybrid cells showed high activity of endogenous alkaline phosphatase (AP); (3) all the hybrid clones were able to form complex embryoid bodies containing derivatives of all the embryonic germinal layers; (4) the hybrid cells contained synchronously replicating X chromosomes, indicating that they were in an active state; and (5) a set of chimeric animals was generated by injecting hybrid cells into BALB/c and C57BL/6J mouse blastocysts. Evidence for chimerism was provided by the spotted coat derived from 129/Ola mice and identification of 129/Ola glucose phosphate isomerase (GPI) in many organs. Thus the results obtained demonstrated that the hybrid cells retain their high pluripotency level despite the close contact of the 'pluripotent' HM-1 genome with the 'somatic' spleen cell genome during hybrid cell formation and the presence of the 'somatic' X chromosome during many cell generations. The presence of HPRT of the somatic partner in many organs and tissues, including the testes in chimeric animals, shows that the 'somatic' X chromosome segregates weakly, if at all, during development of the chimeras. There were no individuals with the 129/Ola genotype among the more than 50 offspring from chimeric mice. The lack of the 129/Ola genotype is explained by the imbalance of the sex chromosomes in the hybrid cells rendering the passage of hybrid cell descendants through meiosis in chimeras impossible. As a result, chimeras become unable to produce gametes of the hybrid cell genotype.
Журнал: Molecular Reproduction and Development
Выпуск журнала: Vol. 50, Is. 2
Номера страниц: 128-138