Optimizing Equilibration Time to Enhance Post-Thaw Viability of Cryopreserved Zebrafish (Danio rerio) Ovarian Fragments.

Histologic and ultrastructural features of cryopreserved ovine ovarian tissue: deleterious effect of 1,2-propanediol applying different thawing protocols

Adding dimethyl sulfoxide (DMSO) to hematopoietic progenitor cells (HPCs) causes an exothermic reaction, potentially affecting their viability. The freezing method might also influence this. The aim was to investigate the effect of 1) precooling of DMSO and plasma (D/P) and white blood cell (WBC)-enriched product, 2) DMSO concentration of D/P, 3) freezing program, and 4) storage method on WBC quality. WBC-enriched product without CD34+ cells was used instead of HPCs. This was divided into six or eight portions. D/P (20 or 50%; precooled or room temperature [RT]) was added to the WBC-enriched product (precooled or RT), resulting in 10% DMSO, while monitoring temperature. The product was frozen using controlled-rate freezing ("fast-rate" or "slow-rate") and placed in vapor-phase or liquid nitrogen. After thawing, WBC recovery and viability were determined. Temperature increased most for precooled D/P to precooled WBC-enriched product, without influence of 20 or 50% D/P, but remained for all variations below 30°C. WBC recovery for both freezing programs was more than 95%. Recovery of WBC viability was higher for slow-rate freezing compared to fast-rate freezing (74% vs. 61%; p < 0.05) and also for 50% compared to 20% D/P (two test variations). Effect of precooling D/P or WBC-enriched product and of storage in vapor-phase or liquid nitrogen was marginal. Based on these results, precooling is not necessary. Fifty percent D/P is preferred over 20% D/P. Slow-rate freezing is preferred over fast-rate freezing. For safety reasons storage in vapor-phase nitrogen is preferred over storage in liquid nitrogen. Additional testing using real HPCs might be necessary.

Objective To evaluate effects of long term storage of semen in liquid nitrogen on the motility, concentration, viability, and abnormality of frozen-thawed. Methods A total of four Friesian Holstein bulls were used for this study. One hundred forty semen straws with produced during period from 2008 to 2013 and stored in the liquid nitrogen at the AI center were used in the research. The sample straw was divided into six groups; each group consist 20 semen straws. For group one all straw semen was produced on the 2013 with storage in liquid nitrogen as long as one year, the group 2, 3, 4, 5, and 6 were produced on the 2012, 2011, 2010, 2009 and 2008 with storage in liquid nitrogen as long as 2, 3, 4, 5, and 6 year, respectively. Results The viability of thawed sperm was not significantly different decreased ( P >0.05) between storage on the 1 year and storage on the 2 years. Whereas, the viability was significantly different ( P P >0.05) on the storage 1, 2 and 3 years. Whereas, the motility was significantly different ( P P >0.05) on the storage 1, 2 and 3 years. Whereas, the abnormality was significantly increased ( P P >0.05) during storage in liquid nitrogen as long as six years. Conclusions Based on the results in these experiments, it may be concluded that concentration sperm during one year storage in liquid nitrogen resulted in similar concentration storage as long as six years. However, the viability and motility sperm thawed storage in liquid nitrogen during six years was lower than storage on the 1 and 2 years.

Complete follicular development and recovery of ovarian function of frozen-thawed, autotransplanted caprine ovarian cortex

Autologous Hematopoietic Progenitor Cell Infusion Adverse Event Associated with Apheresis Product Granulocyte Content

Post-thaw sperm characteristics following long-term storage of boar semen in liquid nitrogen

Effect of reduced glutathione supplementation on cryopreservation induced sperm cryoinjuries in Murrah bull semen

Embryogenic suspension cells of two commercially cultivated aromatic Indica rice varieties, Basmati 385 and Pusa Basmati 1, were cryopreserved using a simple one-step freezing procedure that does not require a controlled-rate freezer. The procedure involves osmotic pre-conditioning of cells with mannitol, addition of a cryoprotectant solution consisting of sucrose, dimethyl sulfoxide, glycerol, proline, and modified R2 medium, cooling to -25°C for 2 h in a freezer, and then storage in liquid nitrogen. After rapid thawing at 45°C, these cultures showed post-thaw cell viability of 5.6 to 10.5% and formed actively dividing, readyto-use cell suspensions in 20-35 d when cultured directly into liquid medium. Plants were regenerated from cell clumps as well as from colonies formed by protoplasts that were isolated from suspension cells re-established from cryopreserved cells, with frequencies higher (54-98%) than, or comparable to, those obtained from three to four-month-old original non-frozen cell cultures. Cell viability and regeneration frequencies of post-thawed Pusa Basmati 1 cultures were similar to those obtained from the suspension cells cryopreserved using the conventional slow-freezing procedure which involves pre-freezing cells to -40°C at the rate of -0.2°C per min prior to immersion in liquid nitrogen. In Basmati 385, however, cells frozen at --25°C showed lower post-thaw cell viability than those preserved using the slow-freezing procedure, but these cells produced cell suspensions that had greater shoot morphogenetic potential. The study indicates the beneficial effect of this simple freezing procedure, not only for preserving desirable cultured cells but also for an enrichment of embryogenic cells.

Survival, pathogenicity, and in vitro growth rate of three isolates of Phytophthora clandestina were tested at monthly intervals for up to 10 months following storage in liquid nitrogen and sterile deionised water on millet seeds at 4°C Storage of cultures in sterile deionised water on millet seeds maintained high recovery rates (75–80%) of all three isolates after 10 months without loss of pathogenicity. The success of liquid nitrogen storage of P. clandestina was variable with only one isolate surviving more than 3 months storage. There was also an effect of the protective solutions used on long term storage and maintenance of pathogenicity. Glycerol (10%) allowed a high recovery rate (89%) of one isolate without loss of pathogenicity for 10 months whilst DMSO (5%) provided a maximum storage period of 3 months with a significant reduction in pathogenicity in the three isolates tested. This study demonstrates that sterile water storage is superior to liquid nitrogen for the medium to long term storage of P. clandestina.

The ability of seed of native Western Australian species to be stored using cryopreservation methods was investigated by subjecting seed of 90 native species representing 84 genera and 33 families to storage in liquid nitrogen. Seed of 68 native Western Australian species were germinated after storage in liquid nitrogen for two weeks following treatments which involved direct plunging into liquid nitrogen or slow cooling at 0.4°C min−1 in 15% dimethyl sulphoxide (DMSO) or slow cooling at 0.4°C min−1 in 35% DMSO. The largest number of species (37) responded positively to direct plunging without pretreatments, with only 10 species responding to slow cooling in 15% DMSO. Thirty one species had enhanced germination and 10 species depressed germination after any of the liquid nitrogen treatments. There were no trends in a species ability to survive liquid nitrogen storage and freezing regime, moisture content, seed size or taxonomic relatedness. However, hard seeded species belonging to the families Caesalpinaceae and Papilionaceae showed a consistently high degree of tolerance to liquid nitrogen storage. Significant physical damage to seed and cotyledons only occurred in Templetonia retusa (Papilionaecae) and this was alleviated by nicking the seed coat. This study indicates that seed of a large proportion of native Western Australian species may be amenable to storage in liquid nitrogen and that at least 40% of the listed rare and endangered species of Western Australia could be maintained in this way.

Glucose/lactate metabolism of cryopreserved intact bovine ovaries as a novel quantitative marker to assess tissue cryodamage

Effect of Long-Term Cryogenic Storage and Conidial Suspending Agents on the Virulence of Beauveria bassiana toward Solenopsis invicta

The viability of 250 basidiomycete strains was determined after a 2-d and then after a 2-year storage under liquid nitrogen using two different freezing protocols. Using an original agar plug protocol (OP), 162 strains (65%) of the 250 strains survived a 2-d storage and 158 strains (63%) survived a 2-year storage in liquid nitrogen. Using a straw protocol (CP), 246 strains (98%) of the 250 strains survived a 2-d storage and 243 strains (97%) a 2-year storage in liquid nitrogen. In addition, other 106 strains were newly estimated using the CP protocol; 104 (98%) of them survived successfully a 2-d storage and 101 (95%) of them survived a 2-year storage in liquid nitrogen. The results indicate that the protocol used for cryopreservation can significantly influence strain survival. Markedly better results were obtained using the CP protocol.

Cryopreservation of freshly isolated synaptosomes prepared from the cerebral cortex of rats

Microbial flora of spermatophores from black tiger shrimp(Penaeus monodon) declines over long-term cryostorage