• Journal of Marine Life Science
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• v.1 no.1
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• pp.70-78
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• 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
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• v.21 no.1
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• pp.125-131
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• 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.

• The Journal of Korean Obstetrics and Gynecology
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• v.32 no.2
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• pp.119-128
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• 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.

• Development and Reproduction
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• v.1 no.1
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• pp.29-36
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• 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
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• v.15 no.1
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• pp.23-33
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• 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
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• v.29 no.3
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• pp.193-199
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• 2005

The cryopreservation of germ cells, sperm and embryos, has been largely used to increase the effect of artificial reproductive techniques for human infertility, but the efficiency of germ cell cryopreservation has been conkoversial till now. Thus, the effect of the cryopreservation of human sperm used for ICSI and the effect of the cryopreservation of embryos produced by ICSI on fertilizatiof development and pregnancy were investigated. Sperm freezing did not affect fertilizatiort development and pregnancy rates. Also, there was no significant difference between ejaculated and testicular sperm in ferclizatiort development and pregnancy. Embryo freezing methods, slow freezing and vitrificatior did not differ each other in viability and pregnncy rates. However, ICSI embryo freezing significantly decreased pregnancy rate compared to fresh embryos freezing (p<0.05). In conclusiof this result suggested that cryopreservation of sperm for ICSI did not affect on the resulted embryo development and pregnancy, but ICSI embryo cryopreservation would significantly inhibit pregnancy.

• Journal of Embryo Transfer
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• v.31 no.4
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• pp.355-365
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• 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.

• Korean Journal of Plant Resources
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• v.17 no.2
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• pp.75-81
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• 2004

This study was conducted to investigate effects of cryopreservation by vitrification on the seed germination rate and growth and physiological properties of seedlings of Maackia amurensis. Cryopreservation significantly decreased the germination rate of seeds of M. amurensis, but the reduction of germination rate was mitigated by the treatment of cryoprotectant (plant vitrification solution, PVS2) before plugging into liquid nitrogen and fast thawing rate after cryopreservation. Long-term PVS2 exposure decreased seed germination rate, whereas cryopreservation time didn't have influence on seed germination rate. In addition, growth and physiological properties of seedlings were not affected by PVS2 exposing time and cryopreservation time. Therefore cryopreservation could be widely used as a technique of long-term ex situ conservation without any damage and deterioration of cells or tissues of the forest seeds. However, in order to increase the effect of cryopreservation, we have to develope the lower toxic cryoprotectant and suitable techniques to the structural or chemical properties of a variety of seeds.

• Restorative Dentistry and Endodontics
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• v.46 no.2
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• pp.26.1-26.15
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• 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.

• Journal of Aquaculture
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• v.12 no.1
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• pp.1-5
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• 1999

To obtain fundamental data for sperm cryopreservation in river puffer (Takifugu obscurus), the proper conditions of cryopreservation were investigated. In the sperm cryopreservation of river puffer, marine fish Rinser's solution (MFRS) was found to be good diluent and dimethyl sulfoxide (DMSO) was proved to be superior to glycerol as a cryoprotectant. The highest fertilization rate was achieved when river puffer sperms were cryopreserved with MFRS adding 5％ DMSO.