|
KMID : 0358419740170080485
|
|
Korean Journal of Obstetrics and Gynecology
1974 Volume.17 No. 8 p.485 ~ p.491
|
|
|
Effects of Steroid Hormoned on the Germinal Vesicle Break-down of Mouse Oocytes In Vitro (Ⅰ) -On Culture Methods and Effects of Progesterone-
|
|
권혁방/Kwon HB
정순오/조완규/Chung SO/Cho WK
|
|
|
Abstract
|
|
|
|
The present studies have been conducted in an attempt to observe the effect of steroid hormoned on the germinal vesicle break-down(GVBD) of the mouse oocytes in vitro. Follicular oocytes were obtained from the ovaried of 3-to 4-week old. A strain female mice. The basic midium was a modified Krebs-Ringer bicarbonate supplemented with 0.4% bovine werum albumin (BSA). Progesterone or estradiol-17β dissolved in absolute ethanol was added to the medium in desired concentration. Maximum concentration of ethanol was not exceeded by 1% of the medium. The oocytes were liberated in the medium by puncturing the follicles with a sharp needle under a dissecting microscope and only those with an intact germinal vesicle were set in midium for culture. The cultivation was performed by applying two different culture sistems in order to compare their utilizability. One is a paraffin oil method which has been well established for embryo culture since development by Brinster (1963) and the other is a micro tube culture system developed by Cho (1974). Ten to twenty oocytes were placed in the medium set either under paraffin oil or in a micro tube and cultured for a period at 37℃ in an incubator supplied with 5% CO2 in moistened air. After culture, the oocytes were fixed in acetic alcohol for 20-24 hours and stained by 0.5% aceto-alcmoid for microscopical observation of nuclear changes. The results were as follows : 1) When we applied the paraffin oil method which allows the direct contact of the medium with the oil through the surface, the germinal vesicle of the oocytes exposed to progesterone (0.1-100㎍/ml) was broken down within 3 hours and the majority of the oocytes the formed metaphase Ⅱ chromosomes in 16 hours in the incubator. It is clear that the paraffin oil method may not be suitable for the investigation on the fat soluble agents such as steroid hormones on the oocytes behaviour, because the agents in the medium may diffuse through paraffin oil during culture, so that the efficiency of the agents would be less. 2) On the other hand, the break-down of the germinal vesicle of the mouse oocytes was almost completely suppressed as long as they were exposed to progesterone (50㎍/ml) in a micro tube which has been developed for the purpose to study the effect of fat soluble agents dissolved in medium on the oocytes or preimplantation embryos. The system is designed to eliminate direct contact of the medium with the oil by imposing air between two in a micro tube, in order to prevent diffusion of the hormone to oil. It becomes apparent that this culture system could be appropriate for evaluation of the effect of the steroid hormones on the oocytes maturation during culture. 3) Estradiol-17β also seemed to inhibit the break-down of the germinal vesicle of the oocytes, but the inhibitory action was far less than that of progesterone. 4) The inhibitory action of the progesterone was not lessened evev though the oocytes were cultured with cumulus cells, or cultured in medium containing an increased dose of energy sources such as calcium lactate or sodium pyruvate.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
 
|
|
|
Listed journal information
|
|
  
|