• Title/Summary/Keyword: 유리화동결

Search Result 81, Processing Time 0.023 seconds

Effect of Ethylene Glycol(EG) and Propylene Glycol(PG) on the Viability of Frozen-thawed Primordial Germ Cells(PGCs) on Korean Native Chicken(Ogye) by Vitrification (한국재래닭(오계)의 유리화 동결 시 생존율에 미치는 Ethylene Glycol(EG)과 Propylene Glycol(PG)의 영향)

  • Kim, Hyun;Kim, Dong Hun;Han, Jae Yong;Choi, Sung Bok;Ko, Yeoung Gyu;Do, Yoon Jung;Seong, Hwan-Hoo;Kim, Sung Woo
    • Korean Journal of Poultry Science
    • /
    • v.40 no.3
    • /
    • pp.197-205
    • /
    • 2013
  • This study established a method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for preservation of the species. The purpose of this study was to compare the effects of Ethylene Glycol (EG) and Propylene Glycol (PG) on viability of cryopreserved PGCs with vitrification in Korean Native Chicken (Ogye), and to fine should be find or to the optimal protocol for PGCs freezing. One of the important components of cryopreservation process is cryopreservation medium that plays a vital role in preventing cellular injury during freeze-thawing. Cryoprotective agents have been known to improve cell viability after freeze-thawing. PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents. Gonads were harvested from stage 28 chick embryos and pooled in groups of 10E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments: 2.5% EG, 5% EG, 10% EG, 2.5% PG, 5% PG, 10% PG, and 0% cryoprotectant as a control. Effects of exposure to vitrification solution and vitrification, with different concentrations of the cryoprotectant solution, were examined. After freezing and thawing, survival rates of the frozen-thawed PGCs from the 0, 2.5, 5, 10 and 15% EG plus FBS treatment were 44.24%, 64.51%, 85.63%, 80.51% and 73.52% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05)(85.63% vs 66.81%). Therefore, these systems may contribute in the improvement of cryopreservation for a scarce species in birds preservation. This study established a method for preserving chicken PGC that enables systematic storage and labeling of cryopreserved PGCs in liquid N at a germplasm repository and ease of entry into a database. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

The Evaluation of Various Conditions in the Cryopreservation of Primordial Germ Cells on Korean Native Chicken (Ogye) (한국재래닭(오계)의 원시 생식 세포의 냉동 보존에 있어서 여러 조건의 평가)

  • Kim, Hyun;Cho, Young Moo;Han, Jae Yong;Choi, Sung Bok;Byun, Mi Jeong;Kim, Young Sin;Ko, Yeoung-Gyu;Seong, Hwan-Hoo;Kim, Sung Woo
    • Korean Journal of Poultry Science
    • /
    • v.41 no.4
    • /
    • pp.249-259
    • /
    • 2014
  • Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps and freezing media on the rates of viability of cryopreserved chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos of Korean Ogye (KO) and Commercial breeds (C), using the MACS method were suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG)). Gonads were harvested from stage 28 chick embryos and pooled in groups of 5, 10, 15, 20E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments : 2.5%, 5%, 10%, 15% and 0% cryoprotectant (DMSO, EG, PG) as a control. Effects of exposure to slow freezing and vitrification, with different concentrations of the cryoprotectant solution, were examined. After vitrification and slow freezing, survival rates of the frozen-thawed PGCs from the 10% EG plus FBS treatment were 85.63%, and 66.14% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05) (85.63% vs 66.81%) by vitrification. This study established a method for preserving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at a germplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

Development of In Vitro Produced Bovine Embryos after Vitrification with Various Containers (다양한 Container로 유리화 동결된 체외생산 소 수정란의 발달에 관한 연구)

  • Lee, Y.J.;Ko, D.H.;H.T., Lee;Chung, K.S.
    • Korean Journal of Animal Reproduction
    • /
    • v.25 no.4
    • /
    • pp.371-379
    • /
    • 2001
  • The objective of this study was to establish an effective cryopreservation method of in vitro-produced bovine embryos. For the vitrification, in virtro-produced embryos at 8-cell, morula and blastocyst stages were exposed to freezing solution containing 5.5 M EG (EG 5.5) for 20 sec, loaded on each containers such as EM grid, OPS and Cryo-loop, and then immediately plunged into liquid nitrogen at -196$^{\circ}C$. Thawed embryos were serially diluted in 0.5, 0.25 and 0.125 M sucrose in m-HPBS, each for 1 min, and cultured in CRlaa medium supplemented with 10% FBS. Significant differences in the rates of re-expanded and hatched embryos were not observed among these embryo containers. The total cell number of expanded blastocyst cultured in vitro after vitrification was examined by Hoechst staining. There were no differences between non-vitrified (180.0 $\pm$ 5.4) and vitrified groups (178.0 $\pm$ 7.5). In addition, when the cellular injuries after vitrification were compared by double staining. There were no significant difference in the ratio of live and dead cells between non-vitrified group (176 : 4) and vitrified group (172 : 6). Therefore, these results suggest that bovine embryos can be cryopreserved easily, effectively and successfully by vitrification using various containers, such as EM grid, OPS or Cryo-loop in the presence of EG 5.5 freezing solution.

  • PDF

A Study for the Changes of The Micro Structure by Deterioration Factors in Concrete for Nuclear Power Plant (원전콘크리트의 열화요인에 따른 미세구조의 변화에 대한 연구)

  • Kim, Do-Gyeum;Lee, Jang-Hwa;Lee, Ho-Jae
    • Proceedings of the Computational Structural Engineering Institute Conference
    • /
    • 2010.04a
    • /
    • pp.766-769
    • /
    • 2010
  • 본 논문은 콘크리트 구조물 중 원전구조물에서 열화요인에 따른 미세구조적 변화에 대해서 평가하였다. 이는 원전구조물의 경우 열화현상이 발생하게 되면 일반 구조물에 비해 심각한 영향을 초래하기 때문에 기존의 열화 평가 방법에 의존하기 보다는 미세구조적 관점에서 콘크리트의 열화를 재평가해야 한다. 그에 일환으로 열화 요인 중 동결 융해와 황산염에 대한 미세구조 평가를 실시하였다. 동결융해의 경우, 미세구조적 관점에서의 미세공극의 양이 증가하는 것을 확인하였으나 그 증가 폭이 크지 않음을 알 수 있었으며, 물리적 실험에서도 그 변화가 매우 작음을 확인할 수 있다. 그리고 황산염에서는 초지 침지 구간에서는 플라이 애쉬를 사용한 원전 콘크리트 배합이 콘크리트에 더 유리하게 작용함을 알 수 있다.

  • PDF