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배양중인 생쥐 여포난자의 핵붕괴에 미치는 Steroid Hormone의 영향 ( I )
권혁방(HB Kwon),정순오(SO Chung),조완규(WK Cho) 대한산부인과학회 1974 Obstetrics & Gynecology Science Vol.17 No.8
생쥐여포난자에 미치는 steroid hormone의 영향을 관찰하기 위하여 두 가지 배양 방법을 사 용하여 배양해 본 결과 다음과 같았다. 1) 종래에 사용되어 오던 paraffin oil method를 사용하면 progesterone을 포함한 배양액 (0.1-100㎍/ml)에서 난자들은 3시간 내에 핵붕괴를 일으키고 그 이후의 성숙과정도 대조군 과 같이 정상적으로 유도되었다. 2) 새로 고안된 micro tube culture method를 사용하면 progesterone(50㎍/ml)을 포함한 배 양액에서는 핵붕괴가 억제되었다. 즉 progesterone은 감수분열을 억제하는 작용을 한다. 3) Steroid hormone과 같은 유용성 물질의 영향을 관찰할 때에는 oil과 배양액의 직접 접촉 이 없는 micro tube culture method를 사용해야 그 효과를 측정할 수 있다. 4) Estrogen도 난자의 성숙을 억제하는 경향이 있으나 progesterone 만큼 완전환 효과를 나 타내지 못한다. 5) 에너지원인 pyruvate 등의 농도를 높여주었어도 progesterone의 효과는 거의 완화되지 않았다. 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.
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문화숙(HS Moon),권혁방(HB Kwon),정순오(SO Chung),곽현모(HM Kwak) 대한산부인과학회 1977 Obstetrics & Gynecology Science Vol.20 No.1
백혈구가 여포난자의 성숙에 미치는 영향을 관찰하고저 배양중인 생쥐의 여포난자에 여러 방법으로 백혈구를 처리해 본 결과 다음과 같은 결론을 얻었다. 1. 백혈구를 포함한 혈장이나, 백혈구를 제거한 혈장에서 난자의 제2차 분열 중기까지의 성숙율은 비교적 좋았다. 2. 3×10⁴cell/㎣ 으로 농축된 백혈구는 난자의 성숙을 현저히 억제한다. 3. 생쥐난자의 이종 백혈구에 대한 감수성은 생쥐의 백혈구에 대한 것과 비슷하나 사람의 백혈구에 대한 저항성이 생쥐의 그것에 보다는 약간 강하다. 4. 파괴시키거나 활동성을 약하게 한 사람의 백혈구나 생쥐의 백혈구를 사용한 결과도 난자 성숙 억제 효과는 줄어 들지 않았다. This experiment was undertaken in order to know the effect of leucocytes on the maturation of mouse oocytes in vitro. Leucocytes obtained from heart puncture of mouse(3×10⁴ cells/㎣) inhibited the maturation of mouse oocytes. The egg toxic activity declined with decreasing leucocyte concentration . It was found that egg toxic effect of leucocytes is not species specific. The activity of intact leucocytes or equal numbers of leucocytes that were destroyed was similar and which seems not to be influenced by the physiological state of leucocytes.
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