scholarly journals CRYOPRESERVATION OF DORMANT GRAPE (Vitis sp.) BUDS.

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1090e-1090 ◽  
Author(s):  
Virgil Esensee ◽  
Cecil Stushnoff ◽  
Philip L. Forsline
Keyword(s):  
Liquid Nitrogen ◽  
Electrolyte Leakage ◽  
Slow Freezing ◽  
First Report ◽  
Vitis Sp ◽  
Direct Immersion ◽  
Backup Storage

There is need for backup storage of clonally propagated plant cultivars of numerous taxa. Initial tests, using a protocol developed for dormant apple buds that includes desiccation and slow freezing prior to immersion in liquid nitrogen (-196 C), was not effective with `Valiant' grape. Accordingly, replicates of V. vinifera `Riesling', V. riparia, `Valiant' and a V. amurensis × riparia cross were also tested for survival at –196 C, following desiccation to 25% & 18% water (fwb) and direct immersion into liquid nitrogan. Visual and electrolyte leakage ratings following nine days of dehydration in moist peat were used to assess viability. Direct immersion of desiccated samples resulted in survival for some buds of `Valiant' and a V. amurensis × riparia cross. V. riparia showed some survival when field hydrated and at 25% water, while all buds desiccated to 18% survived. `Riesling' did not survive desiccation, and was killed by all -196 C treatments. The apple protocol was partially effective, in combination with desiccation to 18% in `Valiant' and V. riparia. This is the first report of grape bud survival in liquid nitrogen and more detailed studies are planned.

Preservation of Morphological Integrity and Clot Retraction Activity of Human Platelets After Freezing

1964 ◽  
Vol 11 (01) ◽  
pp. 222-229 ◽  
Author(s):  
Isaac Djerassi ◽  
Albert Roy ◽  
Jorge Alvarado ◽  
Keyword(s):  
Dimethyl Sulfoxide ◽  
Liquid Nitrogen ◽  
Human Platelets ◽  
Slow Freezing ◽  
Critical Conditions ◽  
Plasma Medium ◽  
Clot Retraction ◽  
Controlled Freezing ◽  
Direct Immersion

SummaryHuman platelets frozen at −195° C (liquid nitrogen) retain their morphological integrity and ability to promote clot retraction when 5% dimethyl-sulfoxide and 5% dextrose are added to the suspending plasma medium. Slow freezing was more effective than direct immersion in the liquid nitrogen. Although similar results may be achieved with dimethylsulfoxide alone with rigidly controlled freezing rates, the addition of sugars may permit freezing under less critical conditions.Dimethylsulfoxyd und 5% Dextrose dem Plasmamilieu hinzugefügt werden. Das langsame Einfrieren ist effektiver als das direkte Eintauchen in flüssigen Stickstoff. Obschon ähnliche Resultate mit Dimethylsulfoxyd allein unter exakter Kontrolle der Einfrierungsgeschwindig-keit erreicht werden können, erlaubt die Zugabe von Dextrose ein Einfrieren unter weniger kritischen Bedingungen.

132 SUCCESSFUL EMBRYO TRANSFER OF IN VIVO-PRODUCED RED DEER (CERVUS ELAPHUS) EMBRYOS AFTER CRYOPRESERVATION BY SLOW FREEZING OR VITRIFICATION

2007 ◽  
Vol 19 (1) ◽  
pp. 183
Author(s):  
J. P. Soler ◽  
G. G. Kaiser ◽  
N. Mucci ◽  
L. B. Ferre ◽  
R. H. Alberio
Keyword(s):  
Liquid Nitrogen ◽  
Embryo Transfer ◽  
Cervus Elaphus ◽  
Red Deer ◽  
Slow Freezing ◽  
Alternative Methods ◽  
Embryo Cryopreservation ◽  
Chi Square ◽  
Cryopreserved Embryos

Multiple ovulation and embryo transfer (MOET) programs for red deer (Cervus elaphus) have been established commercially over the last decade, with embryo cryopreservation being a related practice necessary to enhance the use of valuable genetic information. The aim of this work was to establish alternative methods for red deer embryo cryopreservation by using slow freezing with ethylene glycol (SF–EG) and vitrification by open pulled straw (OPS) methods. After surgical flushing of 18 superstimulated donors, 54 transferable embryos were recovered; 28 were transferred fresh to synchronized recipients and the others were cryopreserved by SF–EG (n = 11) or OPS (n = 15), respectively thawed or warmed, and transferred to recipients. Fresh embryos were maintained in Dulbecco's PBS + 20% cow serum (holding medium, HM) until transfer (maximum 3 h after collection). SF–EG cryopreserved embryos were suspended in HM + 1.78 M EG + 0.1 M sucrose + 4 mg mL−1 BSA. After a 10-min equilibration, embryos were loaded individually into 0.25-mL plastic straws and placed into a −7°C methanol bath chamber. After seeding (5 min later), the straws were cooled from −7 to −35°C at a rate of 0.5°C min. Straws were plunged into and stored in liquid nitrogen. Thawing was performed by placing the straws in a 30°C water bath for 30 s; their contents were drained into HM until transfer. Embryos were vitrified using the OPS method with minor modifications. They were first incubated in HM + 1.78 M EG + 1.3 M DMSO for 3 min and then transferred for 25 s into a vitrification solution of HM + 3.56 M EG + 2.6 M DMSO + 0.5 M sucrose. Each embryo was loaded by touching a 1-µL drop with the straw, which was immediately submerged into and stored in liquid nitrogen. Warming was done by placing the narrow end of the straws into HM + 0.25 M sucrose for 5 min. Embryos were then transferred into HM + 0.15 M sucrose for 5 min and finally to HM until transfer. Both types of cryopreserved embryos were transferred a few hours after collection, immediately after thawing or warming. Before embryo transfer, the presence of corpus luteum (CL) of recipients was confirmed by laparoscopic examination. Each embryo was surgically transferred into the apical extreme of the uterine horn ipsilateral to the CL of one recipient. Pregnancy was determined by ultrasonography 41 days after embryo transfer. The pregnancy rate between groups was compared with the chi-square test (P < 0.05). No statistical differences were found between groups (Table 1). Our results show that both vitrification and slow freezing methods with EG are suitable to cryopreserve red deer embryos. Table 1. Pregnancy rates in recipient hinds after transfer of fresh, vitrified, or frozen red deer embryos

scholarly journals Two-Step Freezing Procedure for Cryopreservation of Rumen Ciliates, an Effective Tool for Creation of a Frozen Rumen Protozoa Bank

2003 ◽  
Vol 69 (7) ◽  
pp. 3826-3832 ◽  
Author(s):  
E. Nsabimana ◽  
S. Kišidayová ◽  
D. Macheboeuf ◽  
C. J. Newbold ◽  
J. P. Jouany
Keyword(s):  
Liquid Nitrogen ◽  
Ice Nucleation ◽  
Slow Freezing ◽  
Nucleation Temperature ◽  
Subzero Temperature ◽  
Equilibration Time ◽  
Rumen Protozoa ◽  
Ciliate Species ◽  
Freezing Procedure ◽  
Holding Temperature

ABSTRACT The present study aimed at the long-term storage of rumen protozoa as living cells in liquid nitrogen. The two-step or interrupted slow freezing procedure was used to cryopreserve six of the dominant species of rumen ciliates isolated from monofaunated animals, Dasytricha ruminantium, Entodinium caudatum, Epidinium ecaudatum caudatum, Eudiplodinium maggii, Isotricha prostoma, and Polyplastron multivesiculatum. We optimized the first step in the interrupted slow freezing procedure, from the extracellular ice nucleation temperature to the holding temperature, and studied the effects of the cooling rates on survival. In addition to the nature of the cryoprotectant (dimethyl sulfoxide), the equilibration temperature and equilibration time (25°C and 5 min, respectively), and the holding time at subzero temperature (45 min) recommended previously (S. Kišidayová, J. Microbiol. Methods 22:185-192, 1995), we found that a holding temperature of −30°C, a cooling rate from extracellular ice nucleation temperature to holding temperature of between 1.2°C/min and 2.5°C/min, depending on the ciliate, and rumen juice as the freezing and thawing medium markedly improved the survival rate. Survival rates determined after 2 weeks in liquid nitrogen were 100% for Isotricha, 98% for Dasytricha, 85% for Epidinium, 79% for Polyplastron, 63% for Eudiplodinium, and 60% for Entodinium. They were not significantly modified after a period of 1 year in liquid nitrogen. Four of the five ciliate species cryopreserved for 8 months in liquid nitrogen successfully colonized the rumen when inoculated into defaunated animals. These results have made it possible to set up a bank of cryopreserved rumen protozoa.

Cryopreservation of Human Nasal Ciliated Epithelium

1996 ◽  
Vol 10 (5) ◽  
pp. 291-298 ◽  
Author(s):  
Bin Yang ◽  
Thomas V. McCaffrey ◽  
Eugene B. Kern
Keyword(s):  
Liquid Nitrogen ◽  
Room Temperature ◽  
Storage Time ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Slow Freezing ◽  
Freezing And Thawing ◽  
Ciliary Function ◽  
Nasal Cilia ◽  
Fast Freezing

The purpose of this study was to compare the effects on human nasal cilia of various freezing and thawing methods in order to determine a reliable method of cryopreservation. Ten samples each were preserved by a slow freezing and a fast freezing method. All samples were stored in liquid nitrogen (–196°C) for 1 week. The frozen samples were thawed by one of two methods: 1) rapid thawing—37°C water bath for 3–4 minutes; and 2) slow thawing—room temperature for 15 minutes. Prefreeze and postthaw ciliary beat frequency (CBF) was measured. The slow freezing and fast thawing method (SFFT) resulted in the best viability. An additional 10 samples preserved using the SFFT method stored at –70°C to –90°C did not retain ciliary function. Thirty ciliated samples preserved by the SFFT method were examined after freezing in liquid nitrogen (–196°C) for 1 week, 2 weeks, and 1 month. There was no significant decrease in CBF after cryopreservation at –196°C for 1 week or 2 weeks (P> 0.05). As the storage time increased to 1 month, postthaw CBF decreased 7.25 ± 0.87% when compared to the prefreeze CBF (P < 0.05). We conclude that human nasal cilia preserved by SFFT at –196°C retain activity for up to 1 month.

scholarly journals Physiological, biochemical, and ultrastructural aspects of Coffea arabica L. seeds under different cryopreservation protocols

2021 ◽  
Vol 45 ◽  
Author(s):  
Madeleine Alves de Figueiredo ◽  
Sttela Dellyzete Veiga Franco da Rosa ◽  
Marcela Andreotti Ricaldoni ◽  
Cristiane Carvalho Pereira ◽  
Stefânia Vilas Boas Coelho ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Water Use ◽  
Coffea Arabica ◽  
Cell Structure ◽  
Cell Structures ◽  
Key Factor ◽  
Physiological Quality ◽  
Ultrastructural Characteristics ◽  
Coffea Arabica L ◽  
Direct Immersion

ABSTRACT Cryopreservation is a technique that may potentially conserve the germplasm of species of the Coffea genus for an indeterminate time. The aim of this study was to evaluate the physiological, biochemical and ultrastructural characteristics of cryopreserved seeds of Coffea arabica L., cultivar Catucaí amarelo IAC 62, which was subjected to different protocols regarding dehydration, precooling, cooling, rewarming and cathode water use. According to each protocol, the seeds were subjected to fast or slow drying to moisture contents of 17 or 20% (wet basis), cooled in different ways, and then immersed in liquid nitrogen for 24 hours. Different rewarming times in a water bath were also used. Physiological, biochemical and ultrastructural analyses were performed on the seeds after the cryopreservation steps. Moisture content at a 17% wb is the key factor for the cryopreservation of Coffea arabica L. seeds, which have better physiological quality and better preserved cell structures. Precooling of coffee seeds before immersion in liquid nitrogen does not provide advantages compared to direct immersion. The rewarming times tested (2, 4, and 6 minutes) and cathode water use did not cause changes in the physiological and biochemical quality or in the cell structures of Coffea arabica L. cryopreserved seeds. The pattern of cell structure observed in all seeds indicates that the damage from cryopreservation is less drastic in the cells of the embryos than in those of the endosperm, with the latter less tolerant to the stresses of dehydration, precooling, and rewarming.

scholarly journals Pear Seeds Retain Viability after Liquid Nitrogen Immersion

HortScience ◽  
2001 ◽  
Vol 36 (6) ◽  
pp. 1121-1122 ◽  
Author(s):  
Barbara M. Reed ◽  
Sara Schwanke ◽  
Rebecca Shala
Keyword(s):  
Liquid Nitrogen ◽  
Vapor Phase ◽  
Seed Viability ◽  
The Other ◽  
Physical Damage ◽  
Triphenyltetrazolium Chloride ◽  
Orthodox Seeds ◽  
Direct Immersion

Cryopreservation in liquid nitrogen (LN) is relatively routine for many small, desiccation-tolerant (orthodox) seeds. Seeds of Pyrus species are considered orthodox but have not been evaluated for LN storage. Seeds of freshly collected P. communis L. (`Bosc') were evaluated for germinability and by TZ staining after exposure to four LN treatments: 1) direct immersion and direct removal; 2) direct immersion and 1 minute in LN vapor phase before removal; 3) 2 minutes in vapor phase before immersion and direct removal; and 4) 2 minutes in vapor phase before immersion and 1 minute in vapor phase before removal. Fresh `Bosc' seed viability evaluated by TZ and greenhouse germination tests remained high (83% to 100%) following four types of LN treatments, compared to the controls (77% to 87%). Differences in `Bosc' seed viability were small and TZ results showed no significant differences among the LN treatments. Direct LN immersion and removal resulted in significantly more greenhouse-germinated `Bosc' seeds than the other treatments and fewer control seeds germinated than any LN treated seeds. Fresh `Bosc' seed cryopreserved at 7.9% moisture exhibited high germinability by both TZ and germination tests. LN exposure caused no physical damage to the seeds. Chemical name used: 2,3,5-triphenyltetrazolium chloride (TZ).

scholarly journals Cryopreservation of Shoot Tips and Pollen of Potato

2017 ◽  
Vol 76 (1) ◽  
pp. 75-80
Author(s):  
Paulina Smyda-Dajmund
Keyword(s):  
Liquid Nitrogen ◽  
Shoot Tips ◽  
Popular Method ◽  
Long Term Storage ◽  
Term Storage ◽  
Direct Immersion

Abstract Cryopreservation is a frequently used method of long-term storage of potato meristems and pollen in liquid nitrogen (LN) in temperature of -196°C. This technique allows for theoretically unlimited storage of potato material. The most popular method of potato shoot tips preservation is cryopreservation by the solidification of liquids without crystallization (vitrification).The best method of pollen conservation is its direct immersion in LN. The successful regeneration after vitrification is genotype-dependent, which require optimization of protocol.

scholarly journals First Report of Grapevine (Vitis sp.) Cluster Blight Caused by Fusarium proliferatum in Russia

Plant Disease ◽  
2020 ◽  
Vol 104 (3) ◽  
pp. 991-991 ◽  
Author(s):  
E. G. Yurchenko ◽  
N. V. Savchuk ◽  
E. V. Porotikova ◽  
S. V. Vinogradova
Keyword(s):  
Fusarium Proliferatum ◽  
First Report ◽  
Vitis Sp

scholarly journals Effect of the Duration of Liquid Nitrogen Storage on the Regrowth of Blackberry Cryopreserved by Droplet Vitrification

2017 ◽  
Vol 66 (1-2) ◽  
pp. 44-50
Author(s):  
Tatjana Vujović ◽  
Đurđina Ružić ◽  
Radosav Cerović
Keyword(s):  
Liquid Nitrogen ◽  
Room Temperature ◽  
Shoot Tips ◽  
Lower Survival Rate ◽  
Droplet Vitrification ◽  
Vitrification Solution ◽  
Long Term Preservation ◽  
Direct Immersion

SummaryIn vitro shoot tips of the blackberry cultivar ‘Čačanska Bestrna’ were cryopreserved using the droplet vitrification technique. Upon loading (30 min) in a solution of 1.9 M glycerol and 0.5 M sucrose, the explants were dehydrated for 40 min on ice with the PVS A3 vitrification solution (glycerol 37.5%, dimethyl sulfoxide 15%, ethylene glycol 15% and sucrose 22.5%) and for 40 min at room temperature with the PVS3 solution (glycerol 50% and sucrose 50%). They were subsequently frozen in individual microdroplets of vitrification solution, by direct immersion in liquid nitrogen (LN), and kept therein for 2, 4, 8 and 24 h. The explant rewarming was performed in an unloading solution (0.8 M sucrose) for 30 min at room temperature. The duration of LN exposure did not exert significant effects on the survival and regrowth of explants in both types of vitrification solutions. The survival and regrowth of cryopreserved shoot tips dehydrated with PVS3 solution ranged between 90–95% and 80–90%, respectively. However, dehydration with PVS A3 resulted in a lower survival rate (80–90%) and a considerably lower regrowth rate (55–65%) of explants. Monitoring the shoots regenerated in the in vitro culture revealed their normal capacity for multiplication and rooting in comparison with the controls, which fully confirms the purpose of cryopreservation in the long-term preservation of plant material.

scholarly journals 113COMPARISON OF TWO ETHYLENE GLYCOL EQUILIBRATION TREATMENTS FOR THE QUICK FREEZING OF IN VITRO-PRODUCED BOVINE EMBRYOS

2004 ◽  
Vol 16 (2) ◽  
pp. 178
Author(s):  
A.C. Nicácio ◽  
R. Simões ◽  
C. Yamada ◽  
H.V.A. Caetano ◽  
M.R.B. Mello ◽  
...  
Keyword(s):  
Ethylene Glycol ◽  
Liquid Nitrogen ◽  
Granulosa Cell ◽  
Cell Monolayer ◽  
Slow Freezing ◽  
Bovine Embryos ◽  
Treatment Groups ◽  
Quick Freezing ◽  
Hatching Rates

The aim of this study was to compare two ethylene glycol (EG) equilibration procedures for the quick freezing of in vitro-produced bovine embryos. Cumulus-oocyte complexes (COCs) were collected from slaughterhouse ovaries. COCs were matured in TCM199 containing 10% of bovine fetal serum, LH, FSH and E2, and fertilized. Presumptive zygotes were co-cultured in TCM199 with a granulosa cell monolayer, at 39°C in humidified atmosphere of 5% CO2 in air. Grade 1, expanded blastocysts (n=761) were selected 7 and 9 days after insemination and randomly distributed to one of eight treatment groups. In Equilibration Procedure 1, embryos were exposed to 10% EG for 5 min, and then to 17%, 22% or 28% EG for 60s (respectively referred to as EG 17, EG 22 and EG 28). In Equilibration Procedure 2, embryos were exposed to the same EG solutions as in Equilibration Procedure 1, but the period of exposure was 10min to 10% EG and 30 s to EG 17, EG 22 and EG 28. In Equilibration Procedure 3 (slow-freezing controls), embryos were exposed to 10% EG for either 5 or 10min and then cryopreserved by slow-freezing method at 1.2°C/min. In all treatment groups, EG solutions were prepared in PBS+0.2% BSA, and embryos were exposed to EG solutions at 22°C. Embryos were loaded into 0.25mL straws and heat-sealed. Straws were cooled in liquid nitrogen vapor for 2min, and then plunged into and stored in liquid nitrogen. Straws were thawed in room temperature air for 10s, and then in 25°C water for 20s. Thawed embryos were diluted by transferring them into 0.5ml of PBS+0.2% BSA+0.3M sucrose for 3min, and then 0.5mL of PBS+0.2% BSA for 3min. Embryos were co-cultured on granulosa cell monolayer in TCM199 and evaluated after 24h for blastocyst re-expansion (EXP), and again at 48, 72 and 96h for hatching (HAT). A total of 724 in vitro-produced bovine blastocysts were used as controls to determine hatching rates. The results are presented in the table. Embryos exposed to 10% EG for 10min (Equilibration Procedure 1) yielded significantly higher rates of blastocyst re-expansion and hatching when compared to embryos exposed for 5min (Equilibration Procedure 2, P&lt;0.05). These results suggest that quick freezing of in vitro-derived bovine embryos may be an alternative to vitrification; however, additional studies are needed to optimize cryopreservation protocols and increase post-thaw survival. This project was supported by FAPESP (01/11266-4) Table 1 Effect of equilibration procedure on in vitro re-expansion and hatching rates of embryos cryopreserved by slow and quick freezing methods

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