[논문]돼지 동결 난포란과 이를 이용한 핵이식 배의 체외발생에 관한 연구

이명헌; 이봉구; 김상근

돼지 동결 난포란과 이를 이용한 핵이식 배의 체외발생에 관한 연구
Developmental Rate of Vitrified Porcine Oocytes and Its Application to NT Embryos Constructed by Microinjection of Fibroblast Cells into Vitrified Oocytes 원문보기

韓國受精卵移植學會誌 = Korean journal of embryo transfer, v.20 no.3, 2005년, pp.207 - 215

이명헌 (국립수의과학검역원) , 이봉구 (중부대학교 애완동물자원학과) , 김상근 (충남대학교 수의과대학)

초록
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본 연구는 돼지 난포란의 동결보존 후 생존성과 난자의 활성화 처리에 따른 체외발생율과 이를 이용한 핵 이식배의 체외발생율을 조사하였다. 활성화 처리된 배는 $5\%$ FBS가 첨가된 NCSU 23 배양액으로 $38.5^{\circ}C$, $5\%\;CO_2$와 $95\%$ air의 조건으로 배양하였다. 1. 난포란을 EDS와 $5\%$ PVP로 동결 후 $10\%$ FBS가 첨가된 NCSU 23 배양액으로 $0{\~}10$시간 배양했을 때 체외발생율은 $36.0\%$로서 대조군인 비동결 난포란의 체외발생율 $46.0\%$에 비해 낮았다. 2. Ethanol과 cyclojexamide로 처리 후 42 및 46시간 배양한 배의 분할율은 각각 $33.3\%$, $36.0\%$ 및 $27.1\%$, $30.0\%$로서 대조군의 $8.8\%$, $11.4\%$에 비해 높게 나타났다. 3. 동결 및 비동결 난포란을 이용한 핵이식 배의 융합율과 발생율 간에는 유의한 차이가 없었다. 4. Ethanol과 cyclojexamide로 활성화 처리한 난자를 이용하여 재구축한 핵 이식배의 발생율은 $2.8\%$, $5.3\%$ 및 $1.5\%$, $2.9\%$로서 대조군의 $0.0\%$, $0.0\%$에 비해 높은 발생율을 나타냈다.

주제어

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제안 방법

This study was conducted to investigate the efficacy of vitrification procedure for the cryopreservation of porcine oocytes and the utilization of vitrified oocytes as recipient cytoplasts for somatic cell NT, and observed that porcine oocytes are activated by ethanol and strontium as evaluated by pronuclear formation, and parthenogenetic development. Single fetal donor cells were deposited into the perivitelline space of vitrified enucleation oocytes, followed by electrical fusion and activation.

대상 데이터

, afterwards transferred to a 20 "L drop VS2 solution, and then quickly added to the EDS solution to expose for 1 minute. The oocytes were sealed in a 1.0mm OPP straw(Vajta et al., 1998a) and placed in a LN2 container. Frozen Oocytes were rapidly thawed in a water bath at 30~35 ℃, and then placed in 0.

데이터처리

The results were expressed by treatment as mean土 SD. For comparison of means, Duncans's multiple verification was performed using SAS package of General Linears Model(GLM) procedures(SAS Institute).

성능/효과

1: When the in vitro developmental rates of the -oocytes after being culture for 0~10 hours vitrified with EDS, 5% PVP, NCSU 23 supplemented with 10% FBS were 50.0%, 22.0 %, respectively. This results were lower than the control group(74.
2. Cleavage rates of embryos activated with ethanol and cyclohexamide were superior for ethanol (33.3% and 36.0%), cyclohexamide(27.7 % and 30.0 %) treatments than that of control(8.8% and 11.4%).
3. The fusion and developmental rates to the blastocyst stage between the NT embryos constructed with the vitrified oocytes were no significant differences, and those constructed with non-vitrified control oocytes.
4. Developmental rates to blastocysts of NT embryos constructed with oocytes activated with ethanol and cyclohexamide were superior for ethanol(2.8% and 5.3%), cyclohexamide(1.5 %, 2.9%) treatments than that of control(0.0% and 0.0%).

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