Cryopreservation of porcine ovarian tissue by vitrification method is a promising approach to preserve genetic materials for future use. However, information is not enough and technology still remains in a challenge stage in pig. Therefore, the objective of present study was to determine possibility of vitrification method to cryopreserve porcine ovarian tissue and to confirm an occurrence of cryoinjuries. Briefly, cryoinjuries and apoptosis patterns in vitrified-warmed ovarian tissue were examined by histological evaluation and TUNEL assay respectively. In results, a damaged morphology of oocytes was detected among groups and the rate was significantly (p < 0.05) lower in vitrification group (25.8%) than freezing control group (67.7%), while fresh control group (6.6%) showed significantly (p < 0.05) lower than both groups. In addition, cryoinjury that form a wave pattern of tissues around follicles was found in the frozen control group, but not in the fresh control group as well as in the vitrification group. Apoptotic cells in follicle was observed only in freezing control group while no apoptotic cell was found in both fresh control and vitrification. Similarly, apoptotic patterns of tissues not in follicle were comparable between fresh control and vitrification groups while freezing control group showed increased tendency. Conclusively, it was confirmed that vitrification method has a prevention effect against cryoinjury and this method could be an alternative approach for cryopreservation of genetic material in pigs. Further study is needed to examine the viability of oocytes derived from vitrified-warmed ovarian tissue.

본 연구는 2017년 국내에서 유통되는 유산균을 함유한 건강기능식품과 가공식품 120건에 대하여 프로바이오틱스 함량, pH와 산도를 분석하였다. 또한, 건강기능식품 중 프로바이오틱스 5개 제품에 대한 보존방법에 따른 변화를 조사하였다. 프로바이오틱스 제품 85건 중 2건이 표시량 이하였고, 가공식품 35건은 모두 적합이었다. 프로바이오틱스 제품의 프로바이오틱스수, pH, 산도 평균은 각각 1.2 × 1010 CFU/g (표시량: 3.4 × 109 CFU/g), 5.35, 1.29%이었으며, 가공식품 중 유산균 함유량 표시 식품 17건의 유산균수 평균은 5.8 × 108 CFU/g이었다. 프로바이오틱스 5제품의 보존온도(−20℃, 4℃, 20℃, 40℃)와 보존기간(1, 3, 6개월) 경과에 따른 프로바이오틱스 함량 변화를 관찰한 결과, 보존기간 1개월에서 6개월 후 프로바이오틱스수 감소율은 평균 59%이었고, 4℃에서 가장 낮은 감소율과 40℃에서 가장 높은 감소율을 나타내었다. 또한, 5개 제품 중 3제품에서 보존기간 1개월에서 3개월 경과 후 40℃에서 70%이상 급격한 균수 감소를 관찰할 수 있었다. 따라서 프로바이오틱스 제품의 보존온도와 보관기간에 따른 변화를 고려할 때 프로바이오틱스 제품은 냉장보관을 권장하며 구입 후 되도록 단기간에 섭취하는 것이 바람직한 것으로 판단된다.

Bovine embryos produced by in vitro maturation, feretilization and development was examined for presevation and transfer. The fertilization medium used BO medium with 5 mM/ caffeine and 10/ heparin and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to 1 cells/ motile sperm during fertilization in vitro. At 8~10 hrs after insemination, the oocytes were transferred into CR1aa medium and cultured for 7 days. Embryos were preserved by vitrification method for transfer. When the embryos of early, blastocyst and expanded blastocyst stages were frozen-thawed, the proportions of embryos with normal morphology 83.6, 88.1 and 85.2%. (중략)