• 한국수정란이식학회지
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• 제9권1호
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• pp.15-21
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• 1994

This experiment was investigated the effect of cell stage of embryos at 48 hours post-insemination On in vitro development of IVF embryos. The ovaries of Korean native cows or heifers were obtained from an abattoir and kept on 25 to 28$^{\circ}C$ and transported to laboratorty within 2 hrs. The oocytes were matured in vitro(IVM) for 24 hrs. in TGM-199 supplemented with 35 $\mu$g/$m\ell$ FSH, 10 $\mu$g/$m\ell$ LH, 1 $\mu$g/$m\ell$ estradiol-17$\beta$ and granulosa cells at 39$^{\circ}C$ under 5% $CO_2$ in air. They were fertilized in vitro(IVF) by epididymal spermatozoa treated with heparin for 24 hrs. , and then the zygotes were co-cultured in vitro(IVC) with bovine oviductal epithelial cells for 7 to 9 days. At 48 hrs. post-insemination, the embryos were classfied into 5 to 8-cell, 3 to 4-cell or 2-cell stage and then were co-cultured in vitro(IVC) with bovine oviductal epithelial cells until the embyos reached blastocyst stage. Embryos developed to blastocyst stage were stained with Hoechst 33342 for cell counting. The embryos of 5 to 8-cell stage at 48 hrs. post-insemination with grade I oocytes were significantly (P<0.05) better developed to blastocysts(63.0%) than 3 to 4-cell(42.0%) and 2-cell stage(2.7%) embryos which delayed in the early cleavage, and those embryos cleaved faster in the very early stage seemed to develop to blastocysts earlier. These results indicate that the embryos cleaved faster at 48 hrs. post-insemination seemed to develop to blastocysts earlier.

• 한국수산과학회지
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• 제19권3호
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• pp.219-226
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• 1986

수산식품단백질의 정확한 품질평가를 위한 새로운 실험 모델을 개발하기 위한 일환으로 전보(Ryu등, 1985)에 이어 6종의 해산 갑각류를 선정하여 비교적 최근에 개발된 four-enzyme digestion technique 및 C-PER assay를 사용하여 이의 품질을 평가하였으며, 또한 이들 technique의 갑각류 단백질에 적용 여부 및 선택성을 검토하였고 그 결과에 영향을 미치는 제요인들을 조사하였다. 시료로 사용된 해산 갑각류는 조단백질이 $85\%$ 이상(건물중량)으로 고급의 단백질원이었으며 이에 조지방 및 조회분을 합하면 $95\%$를 상회하여 이들 세 성분이 주성분이었다. In vitro소화율은 새우류의 경우 $83{\sim}86\%$이었고 어체가 작을 수록 소화율은 높은 반면 trypsin 비소화성물질은 적었다. 생 꽃게육의 소화율은 $80\%$ 정도인 반면, 자숙한 붉은 대게류의 소화율은 $86\%$ 이상으로 자숙에 의한 소화율 증가를 보였고, 부위별로는 집게육의 소화율이 높았다. 일반적으로 생 갑각류의 in vitro 소화율이 낮은 것은 선도저하에 의한 육의 pH변화에 기인된 것으로 생각된다. 새우류 및 게류의 lysinc 함량은 표준 ANRC casein보다 높았으나 다른 필수 아미노산인 Trp, Cys, Met 등은 약간 낮았고 특히 Val, Tyr, Phe 등의 함량은 $50\%$ 정도에 불과하였다. 전반적인 해산 갑각류의 C-PER과 DC-PER은 $2.1{\sim}2.4$정도로 표준단백질 및 다른 어류단백질의 C-PER 및 DC-PER보다 낮았으나 예측소화율은 모두 $90\%$ 이상을 상회하여, 지금까지 알려진 C-PER이 낮은 육단백질의 DC-PER은 훨씬 높다는 일반적인 경향과 상이한 결과를 보여 해산 갑각류 단백질 품질평가시, 소화율은 예측소화율(predicted digestibility) 측정법으로 단백효율비는 보다 정확한 in vitro 소화율 측정법의 개발을 전제로 C-PER technique를 적용함이 바람직할 것 같으며, 보다 정확한 in vitro 소화율 측정에는 선도에 따른 최초의pH, 유리아미노산의 함량 및 TIS 함량 등이 고려된 새로운 model이 개발되어야 할 것으로 생각되었다.

• 대한약학회:학술대회논문집
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• 대한약학회 2005년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.187-189
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• 2005

• 한국가축번식학회지
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• 제20권4호
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• pp.413-421
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• 1997

체외에서 포유동물의 난자와 정자의 배양에 관한 연구는 난자의 성숙과정과 수정현상에 대한 많은 새로운 정보를 제공하였다. 동시에 체외수정의 연구로부터 얻은 결과는 또다른 의문을 제기하였다. 특히 동결융해정액을 이용한 돼지체외수정의 경우 낮은 정자의 침입율과 전핵형성율 및 높은 다정자침입(polyspermy)율은 아직도 해결해야할 문제점으로 남아있다. 돼지난자의 성숙에 관한 연구의 성과는 수정후 낮은 전핵형성율을 개선시켰으나 타동물종에 비하면 아직도 매우 낮게 나타나고 있다. 한편 동결정액의 처리를 위하여 caffeine이나 Ca2+와 같은 물질을 수정용 배지내에 첨가하는 등 수정능력획득의 유기를 위하여 여러 가지 방법이 연구되고 있지만 정자의 침입율은 아직도 낮고, polyspermy의 발생율은 높게 나타내고 있다. 따라서 정자의 침입율을 향상시키고 polyspermy를 억제하기 위하여 난관세포와의 공동배양, 난포액을 첨가한 배양액 내에서 정자의 전배양 및 정자농도의 조절은 매우 효과적인 방법으로 이용되어왔다. 그러나 수정란의 체외생산성 향상과 이와 관련된 연구를 보다 효과적으로 수행하기 위해서는 위에서 지적한 문제에 영향을 미치는 요인에 대한 보다 근본적인 이해가 요구된다.

• 한국동물생명공학회지
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• 제35권1호
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• pp.86-93
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• 2020

The establishment of porcine embryonic stem cells (ESCs) from porcine somatic cell nuclear transfer (SCNT) blastocysts is influenced by in vitro culture day of porcine reconstructed embryo and feeder cell type. Therefore, the objective of the present study was to determine the optimal in vitro culture period for reconstructed porcine SCNT embryos and mouse embryonic fibroblast (MEF) feeder cell type for enhancing colony formation efficiency from the inner cell mass (ICM) of porcine SCNT blastocysts and their outgrowth. As the results, porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days showed significantly increased efficiency in the formation of colonies, compared to those for 7 days. Moreover, MEF feeder cells derived from outbred ICR mice showed numerically the highest efficiency of colony formation in blastocysts produced through in vitro culture of porcine SCNT embryos for 8 days and porcine ESCs with typical ESC morphology were maintained more successfully over Passage 2 on outbred ICR mice-derived MEF feeder cells than on MEF feeder cells derived from inbred C57BL/6 and hybrid B6CBAF1 mice. Overall, the harmonization of porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days and MEF feeder cells derived from outbred ICR mice will greatly contribute to the successful establishment of ESCs derived from porcine SCNT blastocysts.

• Asian-Australasian Journal of Animal Sciences
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• 제29권8호
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• pp.1159-1165
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• 2016

The nutritional value of feed proteins and their utilization by livestock are related not only to the chemical composition but also to the structure of feed proteins, but few studies thus far have investigated the relationship between the structure of feed proteins and their solubility as well as digestibility in monogastric animals. To address this question we analyzed soybean meal, fish meal, corn distiller's dried grains with solubles, corn gluten meal, and feather meal by Fourier transform infrared (FTIR) spectroscopy to determine the protein molecular spectral band characteristics for amides I and II as well as ${\alpha}$-helices and ${\beta}$-sheets and their ratios. Protein solubility and in vitro digestibility were measured with the Kjeldahl method using 0.2% KOH solution and the pepsin-pancreatin two-step enzymatic method, respectively. We found that all measured spectral band intensities (height and area) of feed proteins were correlated with their the in vitro digestibility and solubility ($p{\leq}0.003$); moreover, the relatively quantitative amounts of ${\alpha}$-helices, random coils, and ${\alpha}$-helix to ${\beta}$-sheet ratio in protein secondary structures were positively correlated with protein in vitro digestibility and solubility ($p{\leq}0.004$). On the other hand, the percentage of ${\beta}$-sheet structures was negatively correlated with protein in vitro digestibility (p<0.001) and solubility (p = 0.002). These results demonstrate that the molecular structure characteristics of feed proteins are closely related to their in vitro digestibility at 28 h and solubility. Furthermore, the ${\alpha}$-helix-to-${\beta}$-sheet ratio can be used to predict the nutritional value of feed proteins.

• 한국식품영양과학회지
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• 제14권1호
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• pp.13-22
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• 1985

최대의 in vitro digestibility를 나타내는 조건에서 가열처리한 오징어, 굴, 새우, 명태 및 김을 대상으로 rat-PER, NPR 및 in vitro apparent digestibility를 측정한 결과와 최근 개발된 computed PER(C-PER) 및 discriminant computed PER(DC-PER) technique을 이용하여 계산된 결과를 비교하여, 수산단백질의 품질을 신속하게 측정할 수 있는 가능성을 검토하였다. 오징어, 새우, 멍태의 C-PER은 rat-PER 보다 낮게 계산되었으나, 김은 높게 계산되었으며, 굴은 거의 비슷하게 계산되었다. 또한 이들 시료의 DC-PER은 rat-PER 및 NPR에 C-PER보다 근접된 결과를 보였으며, 단백효율비(PER)와 소화율이 높은 것으로 알려진 굴은 가공저장 중 발생된 산패의 결과로 rat-PER 및 C-PER이 표준단백질인 ANRC casein 보다 훨씬 낮게 계산되었으나, DC-PER은 높게 계산되었다. In vitro 및 in vitro digestibility가 높은 시료의 단백질 품질평가는 DC-PER이 정확하였으며, 품질변화가 심하게 일어난 시료나, 소화율이 낮은 시료는 C-PER technique이 유리할 것으로 생각되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권7호
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• pp.1087-1095
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• 2020

Objective: The present study aimed to evaluate the effects of γ-aminobutyric acid (GABA)-producing bacteria (GPB) on in vitro rumen fermentation and on the growth performance and meat quality of Hanwoo steers. Methods: The effects of GPB (Lactobacillus brevis YM 3-30)-produced and commercially available GABA were investigated using in vitro rumen fermentation. Using soybean meal as a substrate, either GPB-produced or commercially available GABA were added to the in vitro rumen fermentation bottles, as follows: control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB; T3, 2 g/L autoclaved GPB; T4, 5 g/L autoclaved GPB; T5, 2 g/L GABA; and T6, 5 g/L GABA. In addition, 27 Hanwoo steers (602.06±10.13 kg) were subjected to a 129-day feeding trial, during which they were fed daily with a commercially available total mixed ration that was supplemented with different amounts of GPB-produced GABA (control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB). The degree of marbling was assessed using the nine-point beef marbling standard while endotoxin was analyzed using a Chromo-Limulus amebocyte lysate test. Results: In regard to in vitro rumen fermentation, the addition of GPB-produced GABA failed to significantly affect pH or total gas production but did increase the ammonia nitrogen (NH₃-N) concentration (p<0.05) and reduce total biogenic amines (p<0.05). Animals fed the GPB-produced GABA diet exhibited significantly lower levels of blood endotoxins than control animals and yielded comparable average daily gain, feed conversion ratio, and beef marbling scores. Conclusion: The addition of GPB improved in vitro fermentation by reducing biogenic amine production and by increasing both antioxidant activity and NH₃-N production. Moreover, it also reduced the blood endotoxin levels of Hanwoo steers.

• 한국가축번식학회지
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• 제21권3호
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• pp.293-302
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• 1997

Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

• 대한핵의학회지
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• 제9권1호
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• pp.5-9
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• 1975