Developmental potential of cloned mouse embryos reconstructed by a conventional technique of nuclear injection.
The fact that most of the advances in mouse cloning by nuclear transfer originate from research in a limited number of laboratories demonstrates the complexity of the reported technologies. The development of alternative and more simple techniques of nuclear transfer may therefore be of interest. Furthermore, the preimplantation biology of cloned mouse embryos originating from somatic cells has not yet been studied in detail. In the present study, a modified conventional injection (mCI) technique for cloning mice from somatic cells is described. The preimplantation development and morphology of the resulting nuclear transfer embryos in comparison with parthenogenetic embryos and embryos obtained by intracytoplasmic sperm injection (ICSI) under comparable conditions was also studied. Finally, the capacity of nuclear transfer embryos for full-term development was investigated. Eighty-nine per cent of oocytes injected with cumulus cell nuclei under mCI conditions survived and formed zygotes. However, the rate of development of these zygotes to the blastocyst stage was significantly lower (29%) than that of ICSI or parthenogenetic zygotes (95 and 92%, respectively). Cloned blastocysts had a significantly lower mean number of cells in the inner cell mass (9) and trophectoderm (52) and a lower inner cell mass:total cell ratio (14%) than did their counterparts (31, 143 and 18% for ICSI and 21, 92 and 18% for parthenogenetic blastocysts, respectively). This correlated with a significantly higher proportion of dead cells in the cloned blastocysts. The poor quality of cloned blastocysts may explain the low rate of full-term fetal development of somatic mouse clones.
Rybouchkin
A
A
Department of Obstetrics and Gynaecology, Poli 3, Ghent University Hospital, De Pintelaan 185, B-9000, Ghent, Belgium. andrei.rybouchkin@rug.ac.be
Heindryckx
B
B
Van der Elst
J
J
Dhont
M
M
D000818:Animals; D001755:Blastocyst / Q000166:cytology; D019976:Cloning, Organism / Q000379:methods*; D003080:Cold Temperature; D004624:Embryo Transfer; D005314:Embryonic and Fetal Development; D005260:Female; D005312:Fetal Blood; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008845:Microinjections; D053652:Nuclear Transfer Techniques; D010312:Parthenogenesis; D020554:Sperm Injections, Intracytoplasmic