Effects of insulin-like growth factor-I, epidermal growth factor and cysteamine on the in vitro maturation and development of oocytes collected from 6- to 8-week-old Merino lambs

2008 ◽  
Vol 20 (5) ◽  
pp. 570 ◽  
Author(s):  
Jennifer M. Kelly ◽  
David O. Kleemann ◽  
W. M. Chis Maxwell ◽  
Simon K. Walker

To improve the viability of embryos produced in vitro from lamb oocytes, maturation medium was supplemented with insulin-like growth factor-I (IGF-I), epidermal growth factor (EGF), cysteamine, and combinations thereof. Experiment 1 examined the effects of IGF-I supplementation and duration of oocyte maturation on nuclear maturation and embryo development while Experiments 2 and 3 examined the effects of cysteamine and EGF supplementation respectively on embryo development. In Experiment 4, embryo development was examined after maturation with various combinations of supplements. IGF-I supplementation increased cleavage rate (P < 0.05) but its effect on the rate of blastocyst production from original oocytes was variable. Supplementation with IGF-I increased (P < 0.01) the proportion of oocytes at Metaphase II (MII) after 18 h of maturation but not at later times. EGF either alone or combined with IGF-I significantly (P < 0.05) increased cleavage rates compared with other treatment groups but EGF consistently failed to improve blastocyst production rates. Cysteamine improved hatching rates but only when supplemented alone. Maturation of lamb oocytes for 22 h in medium supplemented with 100 ng mL–1 IGF-I and 100 μm cysteamine resulted in the production of 16.0 lambs per donor lamb after embryos were transferred to recipient ewes. It is concluded that EGF and, to a lesser extent, IGF-I, whilst beneficial to initial cleavage, can adversely influence subsequent embryo development. Improvements in embryo viability may more likely be obtained by addressing issues that influence fetal oocyte quality than by modifying in vitro methodology.

1997 ◽  
Vol 9 (6) ◽  
pp. 571 ◽  
Author(s):  
Christopher G. Grupen ◽  
Hiroshi Nagashima ◽  
Mark B. Nottle

The effects of epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) on the in vitro maturation of porcine oocytes were examined. Oocytes obtained from the ovaries of slaughtered prepubertal gilts were matured in modified Medium 199 supplemented with 25% porcine follicular ßuid and gonadotropins, and fertilized in vitro. Oocytes were either xed 16 h later to assess fertilization or cultured for 7 days to assess embryonic development. In Experiment 1, the addition of EGF to maturation medium increased the percentage of meiotically mature oocytes (88% v. 70%; P < 0· 001) but did not affect the proportion of fertilized or cleaved oocytes. Blastocysts derived from oocytes matured in medium supplemented with 10 ng mL-1 EGF had a greater number of cells compared with those of control blastocysts (51·1 ± 5· 1 v. 36·0 ± 3·1; P < 0· 02). In Experiment 2, the addition of IGF-I to maturation medium had no effect on meiotic maturation, fertilization or embryonic development. Our ndings demonstrate that EGF plays an important role in both the meiotic and cytoplasmic maturation of porcine oocytes in vitro.


Development ◽  
1990 ◽  
Vol 110 (1) ◽  
pp. 221-227
Author(s):  
A.N. Corps ◽  
D.R. Brigstock ◽  
C.J. Littlewood ◽  
K.D. Brown

125I-labelled epidermal growth factor (125I-EGF) and 125I-labelled insulin-like growth factor-I (125I-IGF-I) bound to trophoderm cells from pig blastocysts obtained on days 15–19 of pregnancy. Specific binding was detected on freshly isolated cell suspensions and on cells cultured for several days. The binding of 125I-EGF was inhibited by increasing concentrations of EGF, but not by various other growth factors and hormones. Chemical cross-linking of 125I-EGF to its receptors using disuccinimidyl suberate (DSS) revealed a radiolabelled band of relative molecular mass 160,000, similar to that identified as the EGF receptor in other cell types. The binding of 125I-IGF-I was inhibited by both IGF-I and insulin, indicating that the receptors were either type I IGF receptors or insulin receptors. Cross-linking of 125I-IGF-I to serum-free supernatants from trophoderm cultures showed that the cells secreted an IGF-binding protein, giving a complex of relative molecular mass about 45,000. The presence of receptors for EGF and IGF/insulin suggests that these factors could be involved in regulating the growth and development of the early blastocyst.


Reproduction ◽  
1996 ◽  
Vol 107 (1) ◽  
pp. 109-117 ◽  
Author(s):  
P. L. Lorenzo ◽  
P. G. Rebollar ◽  
M. J. Illera ◽  
J. C. Illera ◽  
M. Illera ◽  
...  

1994 ◽  
Vol 14 (7) ◽  
pp. 4588-4595
Author(s):  
D Coppola ◽  
A Ferber ◽  
M Miura ◽  
C Sell ◽  
C D'Ambrosio ◽  
...  

When wild-type mouse embryo cells are stably transfected with a plasmid constitutively overexpressing the epidermal growth factor (EGF) receptor (EGFR), the resulting cells can grow in serum-free medium supplemented solely with EGF. Supplementation with EGF also induces in these cells the transformed phenotype (growth in soft agar). However, when the same EGFR expression plasmid is introduced and overexpressed in cells derived from littermate embryos in which the insulin-like growth factor I (IGF-I) receptor genes have been disrupted by homologous recombination, the resulting cells are unable to grow or to be transformed by the addition of EGF. Reintroduction into these cells (null for the IGF-I receptor) of a wild-type (but not of a mutant) IGF-I receptor restores EGF-mediated growth and transformation. Our results indicate that at least in mouse embryo fibroblasts, the EGFR requires the presence of a functional IGF-I receptor for its mitogenic and transforming activities.


Sign in / Sign up

Export Citation Format

Share Document