scholarly journals In vitro maturation of human germinal vesicle-stage oocytes: role of epidermal growth factor-like growth factors in the culture medium

2010 ◽  
Vol 26 (1) ◽  
pp. 76-81 ◽  
Author(s):  
I. Ben-Ami ◽  
A. Komsky ◽  
O. Bern ◽  
E. Kasterstein ◽  
D. Komarovsky ◽  
...  
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Germinal Vesicle ◽  
Epidermal Growth

scholarly journals In-vitro maturation of human germinal vesicle stage oocytes: role of cumulus cells and epidermal growth factor in the culture medium

1998 ◽  
Vol 13 (6) ◽  
pp. 1638-1644 ◽  
Author(s):  
P. T. Goud ◽  
A. P. Goud ◽  
C. Qian ◽  
H. Laverge ◽  
J. Van der Elst ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Epidermal Growth

256 EFFECT OF EPIDERMAL GROWTH FACTOR ON IN VITRO MATURATION OF CYNOMOLGUS MONKEY (MACACA FASCICULARIS) OOCYTES

2008 ◽  
Vol 20 (1) ◽  
pp. 208
Author(s):  
J. Yamasaki ◽  
J. Okahara-Narita ◽  
C. Iwatani ◽  
H. Tsuchiya ◽  
S. Nakamura ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Germinal Vesicle ◽  
Cynomolgus Monkeys ◽  
Feeder Layers ◽  
Maturation Rate ◽  
Epidermal Growth

Collected oocytes include not only mature oocytes (metaphase II: MII), but also immature oocytes (germinal vesicle: GV, and metaphase I: MI). To establish a dependable artificial indoor breeding program in cynomolgus monkeys, we are planning to carry out in vitro maturation (IVM) using GV and MI oocytes. In this study, we attempted to determine whether different types of feeder layers and epidermal growth factor (EGF) were effective for IVM. Cumulus–oocyte complexes (COCs) were collected from ovaries of 4–10-year-old female cynomolgus monkeys stimulated by the combination of FSH (25 IU kg–1 × 9 days) and hCG (400 IU kg–1) (Torii 2000 Primates 39, 399–406). Oocytes were classified by morphological features: oocytes retaining an intact germinal vesicle nucleus (GV); oocytes that had undergone germinal vesicle breakdown without polar body formation (MI); and oocytes with a first polar body (MII). GV and MI oocytes were co-cultured on monkey cumulus cells (MCC), monkey follicular ovarian cells (MFOC), monkey oviductal cells (MOC), or human solubilized amnion product (HSAP), with TCM-199+10% fetal bovine serum containing epidermal growth factor (EGF; 10 ng mL–1 or 20 ng mL–1). The maturation rate from GV to MII oocytes was 6.7% (MCC), 18.0% (MFOC), 35.7% (MOC), and 28.6% (HSAP) (Table 1). Although higher maturity was observed in MOC and HSAP, the effect of EGF was not found in co-cultures using any feeder layers. The maturation rate from MI to MII oocytes was 33.3% (MCC), 27.8% (MFOC), 55.6% (MOC), and 44.0% (HSAP) (Table 1). The highest maturation rate from GV and MI was observed in co-cultures using MOC. The maturation rate from MI to MII oocytes in the presence of 10 ng mL–1 EGF was 75.0% (MCC) and 73.7% (HSAP) (Table 1), whereas the rate in the presence of 20 ng mL–1 EGF was 59.1% (MCC), 64.3% (MFOC), 92.3% (MOC), and 60.0% (HSAP) (Table 1). Thus, the best maturation rate was a co-culture using MOC as a feeder layer with 20 ng mL–1 EGF. According to our results, maturation rate during IVM depends on the cellular type of feeder layers and the concentration of EGF. EGF is especially effective for maturity from MI to MII oocytes, but not from GV to MI or MII oocytes. Thus, IVM should be carred out under optimal culture conditions, including suitable feeder layer and media plus supplements. In the future, it is important that intracytoplasmic sperm injection be carried out using in vitro-matured MII oocytes for establishment of an artificial indoor breeding program in cynomolgus monkeys. Table 1. Number of matured oocytes co-cultured with different feeder layers and EGF

Functions of epidermal growth factor-like growth factors during human urothelial reepithelialization in vitro and the role of erbB2

2002 ◽  
Vol 30 (4) ◽  
pp. 240-247 ◽  
Author(s):  
Bindels E. ◽  
T. van der Kwast ◽  
Izadifar V. ◽  
Chopin D. ◽  
W. de Boer
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Epidermal Growth

scholarly journals 316EFFECTS OF ESTRADIOL-17² AND PROGESTERONE SUPPLEMENT ON THE RESUMPTION OF MEIOSIS OF CANINE OOCYTES MATURED IN VITRO

2004 ◽  
Vol 16 (2) ◽  
pp. 277
Author(s):  
M.K. Kim ◽  
Y.H. Fibrianto ◽  
H.J. Oh ◽  
G. Jang ◽  
K.S. Lee ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Linear Models ◽  
Uv Light ◽  
Germinal Vesicle ◽  
Domestic Animal ◽  
Estradiol 17Β

In the bitch, oocytes are ovulated at the germinal vesicle (GV) stage and mature in the isthmus of the oviduct around 3 days after ovulation, it is not known what elements trigger the release of this meiotic arrest. Canine IVM has shown limited success with maturation rates, usually around 20% (MII) (Farstad W, 2000 Anim. Reprod. Sci. 60–61, 375–387). Estrogen and progesterone are suggested to play a significant role in causing oocyte resumption of meiosis and progression to MII stage. The purpose of this study was to investigate the role of estradiol-17β (E2) and progesterone (P4) during in vitro maturation of canine oocytes in serum-free tissue culture medium (TCM)-199. Canine oocytes collected from bitches were categorized into three groups based on estrous stages, follicular, luteal, or anestrus, at routine ovariohystrectomy. Oocytes were cultured in vitro in TCM-199 supplemented with E2, P4 or E2+P4 according to experimental design at 39°C in 5% CO2 and O2. After 72h of maturation culture, oocytes were denuded, fixed in a 3.7% paraformaldehyde solution for 10min, stained with Hoechst 33342 in glycerol, and observed under the UV light. Three groups of oocytes were cultured in TCM-199 supplemented with different concentrations (0, 0.1, 1.0 or 2.0μgmL−1) of E2 (Experiment 1, n=898, replications: 5) or P4 (0, 0.5, 1.0 or 2.0μgmL−1, Experiment 2, n=734, replications: 5). Multiple comparisons were implemented using Generalized Linear Models in the SAS 8.12 program. The rates of oocyte maturation to MII stage were higher (P<0.05) in follicular stage oocytes cultured with 2μgmL−1 E2 (17.9%) compared to other supplement groups (0 to 7.6%). No differences (P<0.05) in rate of MII stage oocytes among P4 supplement groups were observed. In Experiment 3, to investigate the combined effects of E2 and P4 on in vitro maturation, three groups of oocytes were cultured in TCM-199 supplemented with 2μgmL−1 E2 and various concentration of P4 (0, 0.5, 1.0 or 2.0μgmL−1, Experiment 3, n=1613, replications: 5). The rate of oocyte maturation to MII stage (11.5%) was higher (P<0.05) in follicular stage oocytes cultured with 2μgmL−1 E2+2.0μgmL−1 P4 supplement compared to other supplement groups (0 to 6.4%). In conclusion, the present study demonstrated that E2 supplement in the culture medium increased maturation of canine oocyte to MII stage and that supplement of P4 alone did not promote oocyte maturation. However, P4 supplemented with E2 further promoted oocyte maturation in the follicular stage compared to E2 supplement alone, indicating that P4 acts synergistically with E2 on canine oocyte maturation in the presence of E2. From our results, we conclude that canine oocytes are exposed to high levels of P4 during maturation due to the preovulatory luteinization of canine follicles which gives rise to high intrafollicular as well as intratubal P4 concentrations-this is very different from the situation in oocytes from other domestic animal species. This study was supported by Biogreen 21-1000520030100000.

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