• 한국해양생명과학회지
• /
• 제1권1호
• /
• pp.70-78
• /
• 2016

Microalgae are of significant importance for future biotechnological applications. Many microalgae banks or laboratories attempt to maintain various microalgae for further research purposes. Cryopreservation has been preferred to reduce a labor-intensive and costly routine sub-culturing. Cryopreservation can also diminish the genetic drift risk. However, cryopreservation as a long term storage of microalgae method are still in developing progress because it cannot be generalized for all microalgae. Microalgae types, cryoprotectant agents (CPAs) types, freezing and thawing methods are the most important factors that should be considered for cryopreservation. In this short review the basic principles and the current advanced of microalgae cryopreservation methods are discussed with a suggested starting parameters for microalgae cryopreservation.

• ALGAE
• /
• 제21권1호
• /
• pp.125-131
• /
• 2006

Long-term preservation of bloom-forming cyanobacteria was evaluated using cryopreservation and freeze-drying of nine strains belonging to four genera and seven species. All test strains, except Aphanizomenon flos-aquae NIER- 10028, showed partial or complete survival following cryopreservation and freeze-drying. Frozen and freeze-dried strains were preserved for more than two years and were revived monthly. Most strains showed higher post-thaw viability after cryopreservation, especially without cryoprotectant compared to freeze-drying. Microcystis aeruginosa NIER-10010, M. viridis NIER-10020, M. ichthyoblabe NIER-10023, M. novacekii NIER-10029 and Oscillatoria sancta NIER-10027 were revived after 2.5 years of cryopreservation. These results suggest that cryopreservation may be an easy and timesaving long-term preservation method for bloom-forming cyanobacteria.

• 대한한방부인과학회지
• /
• 제32권2호
• /
• pp.119-128
• /
• 2019

Objectives: The aim of this case is to report the effects of herbal medicine on a single woman patient with a low level of AMH (anti-$M{\ddot{u}}llerian$ Hormone) in progress of Oocyte Cryopreservation. Methods: A patient with a low level of AMH had symptom of secondary amenorrhea. For preparing oocyte cryopreservation after a long time of secondary amenorrhea, she was treated by twice a day herb medication for 10 months. And we observed the effects of treatments by improvement of symptoms and following up endometrium ultrasonography. After oocyte cryopreservation, for maintaining her menstruation, she was also treated by twice a day herb medication for two and a half months. Results: After treatments, symptom of amenorrhea was improved and the thickness of endometrium was increased as well as AMH in progress of oocyte cryopreservation. So 20 oocytes could be cryopreserved. Conclusions: This case shows that herbal medicine can be a concurrent method for a single woman patient with secondary amenorrhea in progress of oocyte cryopreservation.

• 한국발생생물학회지:발생과생식
• /
• 제1권1호
• /
• pp.29-36
• /
• 1997

Experiments were performed to study the effects of diluents, cryoprotectant, equilibration time, thawing temperature and addition of BSA and egg yolk. Among the various diluents, Alsever's solution was the best for sperm cryopreservation. A combination of Alsever's solution and 15% ethylene glycol showed the better results than others did. Sperm activity indection and survival rate gradually decreased with the equilibration time. The appropriate thawing temperature was 30 ${\pm}1^{\circ}$C. These results indicate that sperm cryopreservation methods can be developed in tiger puffer.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제15권1호
• /
• pp.23-33
• /
• 2007

A simple and reliable cryo technique using desiccation and slow freezing of winter dormant buds was employed for 238 core collection of mulberry germplasm collected from diverse geographical regions and maintained under tropical conditions in the ex situ field gene bank to develop long-term biodiversity conservation for ensuring sustainable utilization of these valuable resources. Desiccation and freezing tolerance of bud grafts and excised shoot apices in the axillary buds of different Morus species under in vivo and in vitro condition indicated species-specific variation and most of the wild Morus species were found sensitive. In vitro regeneration and cryopreservation($-196^{\circ}C$) protocols using differentiated bud meristem like axillary winter dormant buds were worked out for a wide range of Morus species, land races, wild and cultivated varieties. Successful cryopreservation of mulberry winter dormant buds of different accessions belonging to M. indica, M. alba, M. latifolia, M. cathayana, M. laevigata, M. nigra, M. australis, M. bombycis, M. sinensis, M multicaulis and M. rotundiloba was achieved. Among wild species Morus tiliaefolia, and M. serrata showed moderate recovery after cryopreservation. Survival rates did not alter after three years of cryopreservation of different Morus species. ISSR markers were used to ascertain the genetic stability of cryopreserved mulberry, which showed no difference detected among the plantlets regenerated from frozen apices in comparison to the non-frozen material.

• Reproductive and Developmental Biology
• /
• 제29권3호
• /
• pp.193-199
• /
• 2005

인간의 불임을 극복하기 위한 번식공학 기술의 효율성을 증가시키기 위해 성세포의 동결이 널리 수행되고 있으나 동결 기술의 효율성에 있어서 논란의 여지가 있다. 본 연구에서는 체외수정란을 생산하기 위한 난자세포질내 정자미세주입(ICSI) 시술에 사용되는 정자와 이들 기술을 이용 생산한 체외수정란의 동결이 배 발생 및 임신에 미치는 효과를 조사하였다. ICSI방법으로 체외수정란을 생산하는 경우 정자의 동결이 체외수정, 발생 및 임신에 영향을 미치지 않았으며, 특히 동결융해한 사출 및 정소정자에 의한 체외수정율과 발생율 및 임신율도 차이가 없었다. 한편 체외수정란을 동결하는 경우 완만동결과 초자화동결에 의한 체외수정란의 생존율과 임신율은 차이가 없었으나, 동결수정란은 신선수정란에 비하여 임신율이 유의하게 낮았다(p<0.05). 결론적으로 ICSI에 사용되는 정자와 달리 ICSI에 의해 생산된 수정란을 동결하는 경우 임신율을 저하시킬 수 있다.

• 한국수정란이식학회지
• /
• 제31권4호
• /
• pp.355-365
• /
• 2016

The cryopreservation of sperm has become the subject of research for successful artificial insemination technologies. Antifreeze proteins (AFPs), one of the factors necessary for effective cryopreservation, are derived from certain Antarctic organisms. These proteins decrease the freezing point of water within these organisms to below the temperature of the surrounding seawater to protect the organism from cold shock. Accordingly, a recent study found that AFPs can increase the motility and viability of spermatozoa during cryopreservation. To evaluate this relationship, we performed cryopreservation of boar sperm with AFPs produced in the Arctic yeast Leucosporidium sp. AFP expression system at four concentrations (0, 0.01, 0.1, and $1{\mu}g/ml$) and evaluated motility using computer assisted sperm analysis. DNA damage to boar spermatozoa was measured by the comet assay, and sperm membrane integrity and acrosome integrity were evaluated by flow cytometry. The results showed that motility was positively affected by the addition of AFP at each concentration except $1{\mu}g/ml$ (p<0.001). Although cryopreservation with AFP decreased the viability of the boar sperm using, the tail DNA analyses showed that there was no significant difference between the control and the addition of 0.1 or $0.01{\mu}g/ml$ AFP. In addition, the percentage of live sperm with intact acrosomes showed the least significant difference between the control and $0.1{\mu}g/ml$ AFP (p<0.05), but increased with $1{\mu}g/ml$ AFP (p<0.001). Our results indicate that the addition of AFP during boar sperm cryopreservation can improve viability and acrosome integrity after thawing.

• 한국자원식물학회지
• /
• 제17권2호
• /
• pp.75-81
• /
• 2004

본 연구는 산림 수목 종자를 대상으로 다양한 초저온 동결 조건을 적용하고, 초저온 저장 후에 나타나는 종자의 발아 특성 및 유묘의 생장 특성 조사하기 위해 수행되었다. 유리화 과정을 기초로 한 초저온 저장은 다릅나무 종자의 발아율을 크게 감소시켰으나, 초저온 저장 전 동결보호제 처리와 초저온 저장 후의 빠른 해빙은 종자의 발아율 감소를 완화시켰다. 초저온 저장 전 동결보호제 노출 시간은 종자의 발아율에 영향을 주었다. 즉 동결보호제 노출시간이 길어짐에 따라 종자의 발아율은 감소하였다. 그러나 초저온 저장 기간은 종자의 발아율에 영향을 주지 않았다. 또한 동결보호제의 노출 시간과 초저온 저장 기간은 유묘의 생장 특성에 영향을 주지 않았다. 따라서 수목 종자의 초저온 저장은 조직의 손상이나 변형이 없이 장기간 저장할 수 있는 현지 외 보존 기술로 적절하다고 판단된다. 그러나 산림 종자의 보다 효율적인 초저온 저장을 위해서는 각 수종의 종자 특성에 맞는 동결보호제 및 처리 기술이 개발되어야 할 것으로 판단된다.

• Restorative Dentistry and Endodontics
• /
• 제46권2호
• /
• pp.26.1-26.15
• /
• 2021

Objectives: The aim of the present systematic review was to investigate the cryopreservation process of dental pulp mesenchymal stromal cells and whether cryopreservation is effective in promoting cell viability and recovery. Materials and Methods: This systematic review was developed in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement and the research question was determined using the population, exposure, comparison, and outcomes strategy. Electronic searches were conducted in the PubMed, Cochrane Library, Science Direct, LILACS, and SciELO databases and in the gray literature (dissertations and thesis databases and Google Scholar) for relevant articles published up to March 2019. Clinical trial studies performed with dental pulp of human permanent or primary teeth, containing concrete information regarding the cryopreservation stages, and with cryopreservation performed for a period of at least 1 week were included in this study. Results: The search strategy resulted in the retrieval of 185 publications. After the application of the eligibility criteria, 21 articles were selected for a qualitative analysis. Conclusions: The cryopreservation process must be carried out in 6 stages: tooth disinfection, pulp extraction, cell isolation, cell proliferation, cryopreservation, and thawing. In addition, it can be inferred that the use of dimethyl sulfoxide, programmable freezing, and storage in liquid nitrogen are associated with a high rate of cell viability after thawing and a high rate of cell proliferation in both primary and permanent teeth.

• 한국양식학회지
• /
• 제12권1호
• /
• pp.1-5
• /
• 1999

황복(Takifugu obscurus) 정자의 냉동보존을 위한 기초자료를 얻고자, 냉동보존 조건에 관한 연구를 수행하였다. 황복 정자의 냉동보존을 위한 희석액으로는 다른 여러 가지 희석액에 비해 MFRS가 가장 적합하였으며, 동해방지제로는 DMSO가 glycerol에 비해 효과가 좋은 것으로 나타났다. MFRS를 희석액으로 하고 동해방지제인 DMSO를 최종농도가 5%되도록 첨가하여 정자를 냉동보존하였을 때, 해동정자의 수정률은 74.3±7.5%로 가장 보존효과가 좋았다.