title: Effects of donor oocytes and culture conditions on development of cloned mice embryos.
abstract: Mice have been successfully cloned from somatic and embryonic stem (ES) cells using the "Honolulu method." In the present study, different donor oocytes and different culture conditions were compared to evaluate the developmental potential of nuclear transfer embryos reconstructed with an inbred ES cell line HM-1. Oocytes were recovered from two different F1 donors B6D2F1 (C57BL/6 x DBA/2) and B6CBAF1 (C57BL/6 x CBA). There was no effect of oocyte origin on development of cloned embryos to the morulae/blastocyst stage (B6D2F1 44.1% vs. B6CBAF1 45.0%), and the transferred embryos could develop to term. Two culture conditions were compared to show their ability to support development to the morulae/blastocyst stage of reconstructed embryos with B6D2F1 oocytes. The total cell number in the cloned blastocysts cultured in M16 with 20% oxygen was much higher than that observed in CZB with 20% oxygen. Low oxygen concentration during culture of nuclear transfer embryos in CZB medium showed no beneficial effect on pre-implantation development, no embryos developed to term after transfer to surrogate mothers. Our results demonstrated that not only B6D2F1, but B6CBAF1 oocytes, can be used for nuclear transfer. M16 medium is superior for culture of nuclear transfer embryos and low oxygen concentration with CZB medium during culture shows no benefit on development of cloned embryos.
authors.affiliation: Department of Gene Expression and Development, The Roslin Institute, Roslin, Scotland, United Kingdom.
authors.forename: Shaorong
authors.initials: S
authors.lastname: Gao
authors.forename: Michelle
authors.initials: M
authors.lastname: McGarry
authors.forename: Helen
authors.initials: H
authors.lastname: Priddle
authors.forename: Tricia
authors.initials: T
authors.lastname: Ferrier
authors.forename: Bianca
authors.initials: B
authors.lastname: Gasparrini
authors.forename: Judy
authors.initials: J
authors.lastname: Fletcher
authors.forename: Linda
authors.initials: L
authors.lastname: Harkness
authors.forename: Paul
authors.initials: P
authors.lastname: De Sousa
authors.forename: Jim
authors.initials: J
authors.lastname: McWhir
authors.forename: Ian
authors.initials: I
authors.lastname: Wilmut
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D019976:Cloning, Organism; D003470:Culture Media; D046508:Culture Techniques; D004624:Embryo Transfer / Q000662:veterinary; D004622:Embryo, Mammalian / Q000502:physiology*; D005314:Embryonic and Fetal Development; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008811:Mice, Inbred DBA; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D011247:Pregnancy; D013234:Stem Cells / Q000737:chemistry / Q000502:physiology*; D015053:Zygote / Q000166:cytology / Q000254:growth & development
chemical_list: D003470:Culture Media