Development, molecular composition and freeze tolerance of bovine embryos cultured in TCM-199 supplemented with hyaluronan.
Hyaluronan (HA) is glycosaminoglycan that is present from the start of embryonic development and its role and concentration increases with embryo development. The objective of this study was to evaluate if the presence of HA in TCM-199 culture medium had an effect on the development and quality of bovine embryos. There was no effect of HA on the total number of zygotes developing to blastocysts on day 7, however more expanded and hatched blastocyst stages were observed on days 8 and 9 in the group supplemented with HA (p<0.05). Following freeze/thawing, significantly more (p<0.05) embryos cultured in medium supplemented with HA hatched than those cultured in TCM-199 alone or those with BSA. Medium supplemented with HA and BSA significantly increased the level of expression of glucose metabolism Glut-1 gene and embryo compaction Cx43 gene (p<0.05), and had no effect on Glut-5 and IGF-II expression. In addition, HA presence in culture decreased the level of expression of apoptosis Bax and oxidative stress SOX genes (p<0.05). There was significant difference in total number of nuclei between TCM-199 medium only and the remaining media containing BSA or HA plus BSA, between which there was no difference. In summary, our results indicate that the addition of high molecular weight HA to TCM-199 medium that contains BSA on day 4 of culture improved embryo development to hatching and hatched blastocysts and the quality of produced embryos, which were superior to embryos cultured without HA addition.
Palasz
A T
AT
Dpto. de Reproducción Animal y Conservación de Recursos Zoogenéticos, INIA, Madrid, Spain. Palasz@inia.es
Breña
P Beltrán
PB
Martinez
Marcelo F
MF
Perez-Garnelo
S S
SS
Ramirez
M A
MA
Gutiérrez-Adán
A
A
De la Fuente
J
J
D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D002417:Cattle; D003470:Culture Media; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000378:metabolism*; D047108:Embryonic Development / Q000187:drug effects*; D005307:Fertilization in Vitro; D005615:Freezing; D006820:Hyaluronic Acid / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000276:Adjuvants, Immunologic; D003470:Culture Media; D012333:RNA, Messenger; D006820:Hyaluronic Acid