Effects of insulin-transferrin-selenium in defined and porcine follicular fluid supplemented IVM media on porcine IVF and SCNT embryo production.
Insulin-transferrin-selenium (ITS) together has been used in different in vitro maturation system to support in vitro maturation of oocytes. The present study was designed to evaluate the effects of ITS in defined (0.1% PVA) and porcine follicular fluid (10% pFF) supplemented IVM media on the developmental competence of porcine oocytes. Three combinations of ITS, 10 mg/L insulin (Ins), 5.5mg/L transferrin (Tf) and 5 microg/L selenium (Se), 20mg/L Ins, 11 mg/L Tf and 10 microg/L Se, and 30 mg/L Ins, 16.5 mg/L TF and 15 microg/L Se, were used. The data were analyzed by one-way ANOVA and Tukey was used as the post hoc test. Both in the defined and pFF supplemented media, higher concentration of intracellular glutathione was observed in presence of ITS (4.6-4.8, and 6.9-7.1 picomole/oocyte for defined and pFF groups, respectively) compared to the respective control (2.1 and 4.3 picomole/oocyte for defined and pFF group, respectively). ITS decreased polyspermy and increased male pronucleus formation in both the defined and pFF supplemented medium. There was no difference in different treatment groups. The highest frequency of blastocyst formation rate and number of cells in blastocyst following IVF and SCNT was observed in pFF+ITS group (p<0.05). In conclusion, ITS addition during IVM improved the developmental competence of porcine oocytes in both the defined and pFF supplemented groups. Thus, we recommended to supplement porcine IVM medium with 10 microg/mL insulin, 5.5 microg/mL transferrin and 5 microg/mL selenium.
Jeong
Yeon Woo
YW
Sooam Biotech Research Foundation, Bangbae 3 Dong, Seoul 137-851, Republic of Korea.
Hossein
Mohammad Shamim
MS
Bhandari
Dilip P
DP
Kim
Yeun Wook
YW
Kim
Ji Hye
JH
Park
Sun Woo
SW
Lee
Eugine
E
Park
Seon Mi
SM
Jeong
Yeon Ik
YI
Lee
Jong Yun
JY
Kim
Sue
S
Hwang
Woo Suk
WS
D000818:Animals; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D046149:Embryo Culture Techniques / Q000662:veterinary; D004622:Embryo, Mammalian; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D015571:Follicular Fluid / Q000737:chemistry / Q000502:physiology*; D005978:Glutathione / Q000032:analysis; D007328:Insulin / Q000494:pharmacology*; D008297:Male; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D009866:Oogenesis / Q000187:drug effects; D011247:Pregnancy; D018873:Pregnancy Rate; D012643:Selenium / Q000494:pharmacology*; D013552:Swine / Q000196:embryology; D014168:Transferrin / Q000494:pharmacology*
D003470:Culture Media; D007328:Insulin; D014168:Transferrin; D005978:Glutathione; D012643:Selenium