Increasing glucose in KSOMaa basal medium on culture Day 2 improves in vitro development of cloned caprine blastocysts produced via intraspecies and interspecies somatic cell nuclear transfer.
This study was conducted to evaluate the efficiency of potassium simplex optimization medium with amino acids (KSOMaa) as a basal culture medium for caprine intraspecies somatic cell nuclear transfer (SCNT) and caprine-bovine interspecies somatic cell nuclear transfer (iSCNT) embryos. The effect of increased glucose as an energy substrate for late stage development of cloned caprine embryos in vitro was also evaluated. Enucleated caprine and bovine in vitro matured oocytes at metaphase II were reconstructed with caprine ear skin fibroblast cells for the SCNT and iSCNT studies. The cloned caprine and parthenogenetic embryos were cultured in either KSOMaa with 0.2 mM glucose for 8 days (Treatment 1) or KSOMaa for 2 days followed by KSOMaa with additional glucose at a final concentration of 2.78 mM for the last 6 days (Treatment 2). There were no significant differences in the cleavage rates of SCNT (80.7%) and iSCNT (78.0%) embryos cultured in KSOMaa medium. Both Treatment 1 and Treatment 2 could support in vitro development of SCNT and iSCNT embryos to the blastocyst stage. However, the blastocyst development rate of SCNT embryos was significantly higher (P < 0.05) in Treatment 2 compared to Treatment 1. Increasing glucose for later stage embryo development (8-cell stage onwards) during in vitro culture (IVC) in Treatment 2 also improved both caprine SCNT and iSCNT embryo development to the hatched blastocyst stage. In conclusion, this study shows that cloned caprine embryos derived from SCNT and iSCNT could develop to the blastocyst stage in KSOMaa medium supplemented with additional glucose (2.78 mM, final concentration) and this medium also supported hatching of caprine cloned blastocysts.
Animal Biotechnology-Embryo Laboratory, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Federation of Malaysia.
P J
PJ
Kwong
R B
RB
Abdullah
W E
WE
Wan Khadijah
D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D001755:Blastocyst / Q000187:drug effects* / Q000502:physiology; D002417:Cattle; D019976:Cloning, Organism / Q000662:veterinary; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D046149:Embryo Culture Techniques / Q000379:methods* / Q000662:veterinary; D047108:Embryonic Development / Q000187:drug effects* / Q000502:physiology; D005260:Female; D005947:Glucose / Q000494:pharmacology*; D006041:Goats / Q000196:embryology / Q000502:physiology; D006822:Hybrid Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D006824:Hybridization, Genetic / Q000187:drug effects / Q000502:physiology; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D011188:Potassium / Q000494:pharmacology; D013045:Species Specificity; D013997:Time Factors
D000596:Amino Acids; D003470:Culture Media; D005947:Glucose; D011188:Potassium