title: The effects of manipulation medium, culture system and recipient cytoplast on in vitro development of intraspecies and intergeneric felid embryos.
abstract: The aim of this study was to investigate if reconstructed felid embryos obtained by intraspecies or intergeneric cloning can develop in vitro. Fibroblast cells (f) from a domestic cat (DCf), marbled cat (MCf) and bovine (Bf) were used as donor cells, and oocytes (o) from domestic cats (DCo) and bovine (Bo) were used as recipient cytoplasts. There were two intraspecies (donor cell + recipient cytoplast: DCf + DCo and Bf + Bo) and three intergeneric (MCf + DCo, DCf + Bo and MCf + Bo) cloning groups in the study. In Experiment 1, the effects of manipulation media, modified TCM-199 (199H) or Emcare holding medium (EHM), on in vitro development of DCf + DCo embryos were investigated. The blastocyst formation rate (BFR) of the embryos manipulated in EHM (33.3%) was higher (P<0.05) compared with those manipulated in 199H (18.1%). In Experiment 2, DCf + DCo and MCf + DCo embryos were cocultured with or without domestic cat oviductal epithelium cells. Irrespective of coculture, the same BFR was obtained for DCf + DCo embryos (44.4 vs. 38.0%), while MCf + DCo embryos could not develop beyond the morula stage. In experiment 3, although the development of MCf + DCo and DCf + Bo embryos was arrested at the morula stage, 8.6% of MCf + Bo embryos were able to develop to the blastocyst stage. These results demonstrated that EHM was superior to 199H as an embryo manipulation medium and that the DCo and Bo could support the early embryonic development of intergeneric cloned marbled cat embryos up to the morula stage. However, postimplantation development still needs to be investigated.
authors.affiliation: Embryo Technology and Stem Cell Research Center, School of Biotechnology, Suranaree University of Technology, Nakhon Ratchasima, Thailand.
authors.forename: Sumeth
authors.initials: S
authors.lastname: Imsoonthornruksa
authors.forename: Chanchao
authors.initials: C
authors.lastname: Lorthongpanich
authors.forename: Anawat
authors.initials: A
authors.lastname: Sangmalee
authors.forename: Kanokwan
authors.initials: K
authors.lastname: Srirattana
authors.forename: Chuti
authors.initials: C
authors.lastname: Laowtammathron
authors.forename: Wanchai
authors.initials: W
authors.lastname: Tunwattana
authors.forename: Wachiravit
authors.initials: W
authors.lastname: Somsa
authors.forename: Mariena
authors.initials: M
authors.lastname: Ketudat-Cairns
authors.forename: Takashi
authors.initials: T
authors.lastname: Nagai
authors.forename: Rangsun
authors.initials: R
authors.lastname: Parnpai
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002415:Cats; D002417:Cattle; D019976:Cloning, Organism / Q000379:methods*; D003470:Culture Media; D047108:Embryonic Development; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D006824:Hybridization, Genetic; D009028:Morula / Q000166:cytology; D009865:Oocytes / Q000166:cytology / Q000254:growth & development
chemical_list: D003470:Culture Media