배양액과 산소농도가 돼지난포란의 체외성숙과 배발달에 미치는 영향 = Effects of Different Media and Oxigen Concentrations on In Vitro Maturation and Development of Porcine Follicular oocytes

다국어 초록 (Multilingual Abstract)

The present study was carried out to examine the effect of four different media (NCSU (North Carolina State University)-23, PZM (Porcine Zygotes Medium)-3, PZM-4 and TCM (Tissue Culture Medium)-199) and two oxygen concentrations (39 , 5% O2, 5％ CO2 and 90% N2, 5％ CO2 in air) on in vitro production of porcine IVM/IVF embryos.The results were summarized as follows:1) The rates of GVBD and nuclear maturations were not significantly different (p>0.05) for 44 hours of culture with four media in two oxygen concentrations.2) The rates of polyspermy, penetrated sperm(s) and male and female prouclei formation were not significantly different (p>0.05). among four media in two oxygen concentrations. 3) The cleavage rates were not significantly different (p>0.05) among four media in two oxygen concentrations. At day 7 under gas atmosphere of 5% O2, 5％ CO2 and 90% N2, the blastocyst formation was significantly higher (p<0.05) in PZM-3 (19.9±2.4) than other media. Also, NCSU-23 medium gave high rate of blastocyst formation at day 7 under gas atmosphere of 5％ CO2 in air (p<0.05).4) Based on the result of differential staining of porcine blastocyst at dat 7, inner cell mass cell and total cell numbers were not significantly different (p>0.05) among four media in two oxygen concentrations. However, the observed total cell number was higher in PZM-3 medium (36.8±6.5) than other madia.In conclusion, these results suggested that in vitro production of porcine embryos in PZM-3 medium under a gas atmosphere of 5% O2, 5％ CO2 and 90% N2 was effective on the blastocyst formation rate and total blastocyst cell number.

국문 초록 (Abstract)

본 실험은 돼지체외수정란을 생산하는데 적합한 배양액과 산소조건을 구명하기 위하여 NCSU(North Carolina State University)-23, PZM(Porcine Zygotes Medium)-3, PZM-4 및 TCM(Tissue Culture Medium)-199 배양액과 5% 산소조건(39 , 5% O2, 5％ CO2, 90% N2 및 포화습도) 및 20% 산소조건(39 , 5％ CO2 및 포화습도)에서 돼지 미성숙난포란의 체외성숙과 체외배발달을 실시하였다. 본 연구에서 얻어진 결과를 요약하면 다음과 같다.1) 돼지 난포란을 5 및 20% 산소조건하에서 NCSU-23, PZM-3, PZM-4 및 TCM-199 배양액에 44시간동안 성숙을 유도한 결과 난핵포붕괴율과 핵성숙률에 있어서 산소조건 및 배양액간 유의적(p>0.05)차이는 없었다.2) 체외수정을 실시한 결과 정자침투율, 자·웅전핵형성률, 다정자 침입률 및 평균정자수에 있어서 산소조건 및 배양액간에 유의적인 차이가 없었다.3) 체외수정을 실시한 후, 5 및 20% 산소조건의 NCSU-23, PZM-3, PZM-4 및 TCM-199 배양액에서 배양하여 배발달에 미치는 영향을 조사한 결과, 난할률에 있어서는 유의적(p<0.05)인 차이가 없었으며, 배발달 7일째 배반포기 발달률에 있어서 5% 산소조건에서는 PZM-3 배양액이, 20% 산소조건에서는 NCSU-23 배양액이 유의적(p<0.05)으로 높은 결과를 나타냈다.4) 배발달 7일째 배반포기 배의 세포수를 조사한 결과, 내부세포괴세포수와 총세포수는 산소조건과 배양액간에 통계적인 유의성이 인정되지는 않았으나, 5% 산소조건에서 PZM-3 배양액(36.8±6.5개)이 다소 높은 총세포수를 나타냈다.위의 결과를 종합하면 돼지체외수정란의 생산에는 5% 산소조건에서 PZM-3 배양액에서 배발달을 유도하는 것이 배반포기 발달률과 세포수에 있어 효과적이라고 판단된다.

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