Efficiency of SCNT buffalo (Bubalus bubalis) embryos in different culture medium and analysis of mRNA expression of insulin-like growth factors during embryogenesis.
Growth factors in culture media are known to affect the embryo production rates in in vitro production cultures. To improve the efficiency of somatic cell nuclear transfer (SCNT) derived embryos in Indian buffaloes (Bubalus bubalis), embryos were cultured in three different culture mediums viz. Group-A; TCM-199 + FBS, Group-B; TCM-199 + Poly vinyl alcohol (PVA) and Group-C; CR1aa + BSA. Embryo production rate and expression level of insulin-like growth factor genes (IGF-1, IGF-1R, IGF-2 and IGF-2R) were analysed in embryo culture. Cleavage and blastocyst production rates were 62.5% and 22.3% in Group-A, 53.8% and 13.0% in Group-B and 62.0% and 19.2% in Group-C respectively, whereas in in vitro fertilization (IVF) control cultured in TCM-199 plus 10% FBS, rates were 79.1% and 29.4%. Relative gene expression of SCNT embryos was compared with that in IVF control. IGF-1 and IGF-2 mRNA expression at blastocyst stage was up-regulated (p ≤ 0.05) in all culture groups, while IGF-1R and IGF-2R expression was down regulated (p ≤ 0.05) in Group-B and Group-C. In conclusion, the higher mRNA levels at certain stages in different culture conditions affected in vitro development of SCNT embryos. These results show that the transcript level of the insulin-like growth factor genes was significantly altered by in vitro culture condition. Culture medium TCM-199 with 10% FBS produced higher number of embryos and was able to co-op with gene expression of IVF control. Differences continue to be observed between SCNT cultured and IVF embryos, and until these differences are minimized, aberrations in SCNT embryonic development will continue to arise.
Pandey
A
A
Transgenic Research Laboratory, National Bureau of Animal Genetic Resources, Karnal, India.
Gupta
S C
SC
Singh
N
N
Rana
J S
JS
Gupta
N
N
D000818:Animals; D002020:Buffaloes / Q000196:embryology*; D019976:Cloning, Organism; D003470:Culture Media / Q000737:chemistry*; D046149:Embryo Culture Techniques / Q000662:veterinary; D047108:Embryonic Development / Q000502:physiology; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D013002:Somatomedins / Q000235:genetics / Q000378:metabolism*
D003470:Culture Media; D012333:RNA, Messenger; D013002:Somatomedins