Cytogenetic analysis of human somatic cell haploidization.
Despite recent interest in the derivation of female and male gametes through somatic cell nuclear transfer, there is still insufficient data on chromosomal analysis of these gametes resulting from haploidization, especially involving a human nuclear donor and recipient oocytes. The objective of this study was to investigate the fidelity of chromosomal separation during haploidization of human cumulus cells by in-vitro matured human enucleated MII oocytes. A total of 129 oocytes were tested 4-7, 8-14, or 15-21 h after nuclear transfer (NT) followed by electro-stimulation, resulting in 71.3% activation efficiency on average. Haploidization was documented by the formation of two separate groups of chromosomes, originating from either polar body/pronucleus (PB/PN), or only 2PN, which were tested by 5-colour FISH, or DNA analysis for copy number of chromosomes 13, 16, 18, 21, 22 and X. Two PN were formed more frequently than PB/PN, irrespective of incubation time. In agreement with recent reports on mouse oocytes, as many as 90.2% of the resulting haploid sets tested showed abnormal chromosome segregation, suggesting unsuitability of the resulting artificial gametes for practical application at the present time.
Galat
V
V
Reproductive Genetics Institute, 2825 North Halsted Street, Chicago, IL 60657, USA.
Ozen
S
S
Rechitsky
S
S
Kuliev
A
A
Verlinsky
Y
Y
D002467:Cell Nucleus / Q000502:physiology; D020090:Chromosome Segregation / Q000502:physiology*; D020732:Cytogenetic Analysis; D004558:Electric Stimulation; D019075:Electrophoresis, Capillary; D006238:Haploidy; D006801:Humans; D008540:Meiosis / Q000502:physiology*; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D027724:Reproductive Techniques, Assisted; D013997:Time Factors