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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Day 7 cow embryos were frozen in 1.5 M-DMSO in PBS at 0.3 degrees C/min to -36 degrees C and at 0.1 degrees C/min between -36 and -60 degrees C before being plunged directly into liquid nitrogen. They were subsequently thawed (rapidly to -50 degrees C, at 4 degrees C/min from -50 to -10 degrees C, and rapidly again) to room temperature. Embryonic viability was tested by four different transfer techniques. Maximum pregnancy rate (8/12) was obtained with surgical transfer immediately after thawing and dilution of DMSO.
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PMID:The viability of deep-frozen cow embryos. 63 24

Preimplantation stage (16-celled and morula) rabbit embryos were successfully frozen to -196 degrees C. The cooling rate (from a room temperature to 0 degrees C), the presence of the mucin layer surrounding embryos, the ice-seeding treatment and the thawing procedure were examined to determine their effects on the survival of the frozen embryos of Japanese white, New Zealand white and Dutch-Belted rabbits. A high proportion (51%; 16-celled, 69%; morula) of Dutch-Belted rabbit embryos developed in vitro, when they were frozen to -196 degrees C, applying the ice-seeding at -4 degrees C in the presence of 12.5% DMSO, after being cooled to 0 degrees C at the rate of 7-9 degrees C/min, and were diluted by a stepwise addition of 4 different strength PBS on thawing. The highest rate of in vitro development (81%; Japanese white, 75%; New Zealand white, 82%; Dutch Belted embryos) was obtained when the morula stage embryos were frozen to -196 degrees C applying seeding at -4 degrees C after being cooled to 0 degrees C at the rate of 1 degrees C/2.5 min and were diluted, on thawing, by stepwise addition of 6, 3 and 1% DMSO solution and a culture medium. No great difference was found in the survival rate between the embryos covered with the mucin layer and those which had not the coat. All the embryos frozen without applying seeding treatment failed to develop in vitro after being thawed and diluted. Nine out of 27 does each of which received 6 reimplantations of the embryos frozen-thawed became pregnant and were found to be carrying 37 normal fetuses on the 12th day of pregnancy.
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PMID:Survival of 16-celled and morula stage rabbit embryos frozen to -196 degrees C,. 103 62

The discovery of glycerol as an effective cryoprotectant for spermatozoa led to research on cryopreservation of embryos. The first successful offspring from frozen-thawed embryos were reported in the mouse and later in other laboratory animals. Subsequently, these techniques were applied to domestic animals. Research in cryopreservation techniques have included studies concerning the type and concentration of cryoprotectant, cooling and freezing rates, seeding and plunging temperatures, thawing temperatures and rates, and methods of cryoprotectant removal. To date, successful results based on pregnancy rates have been obtained with cryopreserved cow, sheep, goat, and horse embryos but no success has been reported in swine. Post-thaw embryo survival has been shown to be dependent on the initial embryo quality, developmental stage, and species. The freezing techniques most frequently used in research and by commercial companies are identified as "equilibrium" cryopreservation. In this technique the embryos are placed in a concentrated glycerol solution (1.4 M in PBS supplemented with BSA) at room temperature and the glycerol is allowed to equilibrate for a 20-min period. During the cooling process the straws are seeded (-4 to -7 degrees C) and cooling is continued at a rate of 0.3 to 0.5 degree C/min to -30 degrees C when bovine embryos may be plunged into LN2. Sheep embryos are successfully frozen with ethylene glycol (1.5 M) or DMSO (1.5 M) rather than with glycerol. Horse embryos have been frozen in 0.5 rather than 0.25 cc straws but with cooling rates and seeding and plunging temperatures similar to those used with bovine embryos. Swine embryos have shown a high sensitivity to temperature and cryoprotectants probably due to their high lipid content and a temperature decrease to 15 or 10 degrees C causes a dramatic increase in the percentage of degenerated embryos. However, a recent study has shown that hatched pig blastocysts survived exposure below 15 degrees C. Recent research has shown that embryos may also be frozen by a "nonequilibrium" method. This rapid freezing by vitrification consists of dehydration of the embryo at room temperature by a very highly concentrated vitrification media (3.5 to 4.0 M) and a very rapid freeze that avoids the formation of ice allowing the solution to change from a liquid to a glassy state. Vitrification solutions consist of combinations of sucrose, glycerol, and propylene glycol. With this technique, 50% pregnancy rates have been reported with the bovine blastocyst.
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PMID:Status of cryopreservation of embryos from domestic animals. 160 26

Spermatozoa from cauda epididymis of mature mice were suspended in preservation solution (Dulbecco's PBS containing raffinose in combination with glycerol, DMSO or skim milk as freezing protective agents). The suspension was frozen by the dry ice-alcohol method and preserved for 1-120 days in liquid nitrogen (-196 degrees C). Highest sperm viability after thawing was obtained with a combination of 10% raffinose and 5% glycerol or with a combination of 10% raffinose and 10% DMSO. These frozen thawed sperm were found to have fertilizing capacity when used for in vitro fertilization. The 2-cell embryos obtained through the above procedures developed into normal pups at a high rate when transferred into the oviducts of pseudopregnant female mice.
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PMID:[Production of normal young following transfer of mouse embryos obtained by in vitro fertilization using cryopreserved spermatozoa]. 230 89

Murine embryos of mice of four different inbred strains and one hybrid strain were evaluated for their ability to survive quick freezing by post-thaw in vitro development. The embryos were transferred to an equilibration medium [10% 1,2-propanediol and 20% glycerol in modified PBS (mPBS)] for 10 minutes and frozen in a vitrification medium (25% glycerol and 25% 1,2-prapanediol in mPBS) by direct lowering into liquid nitrogen. Following thawing at 30 degrees C, dilution in 1 M sucrose in mPBS and washing in mPBS the embryos were cultured, and development was evaluated 24-28 hours later. The number of fertilized eggs obtained by superovulation differed among the strains. The survival rates evaluated by in vitro cultivation of the post-thawed inbred embryos varied from 50-85% depending on the genotype, whereas the normal live offspring from transfer of frozen-thawed embryos to recipient females confirms that the quick freezing method is an applicable method for storage of genetically defined mouse strains and stocks. The quick freezing technique was applied on 4- and 8-cell (day-3) mouse embryos of hybrids. The in vitro development of frozen thawed 4- and 8-cell embryos (23% and 21% respectively) was found to be significantly lower than that of frozen thawed morulae (89%). Permeation in glycerol-solutions before equilibration significantly increased survival of 4- and 8-cell embryos (66% and 77% respectively). By the use of dimethylsulfoxid (DMSO) in the permeation solutions an even higher survival rate was obtained in the cryopreservation of 8-cell mouse embryos (95%).(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Quick freezing of mouse embryos: freezing of inbred strains and 2- and 4-cell embryos by vitrification. 297 55

As determined by luciferase-luciferin, we recently found that the H2-blocker CIM considerably increased the ATP release from fMLP-stimulated PMN. This observation correlates well with our previous [1] regarding the enhancement of superoxide output (chemiluminescence) in in human neutrophils. by CIM plus fMLP. In order to compare the ATP release from PMN of different donors, a standard procedure has been developed consisting of the determination of the ATP present initially in the cell suspension (without stimulation), ATP release after stimulation with fMLP, and ATP release in the presence of CIM plus fMLP. The whole ATP content per neutrophil was determined after ultrasonication of the cells as well. The mean value of the initially present ATP was 0.45 x 10(-17) mol/cell in the suspension. Stimulation with fMLP plus CIM yielded within 5-10 minutes considerably higher ATP amounts than fMLP alone. The corresponding and statistically significantly different mean values were 2.46 x 10(-17) mol/PMN (s.d. = 1.047) and 1.38 x 10(-17) mol/PMN (s.d. = 0.55), respectively. The whole ATP per neutrophil was found to be 1.22 x 10(-15) mol (mean; s.d. = 0.60) and thus, the stimulation with CIM plus fMLP released about 2.0 per cent, with fMLP alone about 1.0 per cent of the whole ATP. CIM without fMLP did not enhanced the ATP release during the reaction time applied. On the other hand, fMLP-stimulated, lucigenin-amplified chemiluminescence determinations were carried out in the presence of CIM as well; contrarily to our previous method, CIM was dissolved in PBS without DMSO, because DMSO inhibited the chemiluminescence slightly.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Enhancement by cimetidine of chemotactic peptide-stimulated ATP release and chemiluminescence in human neutrophils. 317 91

We have studied the methods of cryopreservation and dilution of the unfertilized and fertilized eggs of ICR mouse. To improve the survival rate, we examined 8 types of methods with various combination of cryoprotectants, dilutions and cooling rates. The highest survival rate for unfertilized eggs was obtained when 1.5 M dimethylsulfoxide (DMSO) + 0.25 M sucrose was added to PBS as a cryoprotectant and when the eggs were diluted by adding PBS with 0.5 M sucrose at room temperature at 5-minute intervals in 5 steps. The total survival rate was 45.4% (p less than 0.01). In the case of fertilized eggs, the highest survival rate (72.6%) was obtained with 10% glycerol + 0.25 M sucrose as a cryoprotectant and a One-step dilution method. The most effective cooling rate was 0.3 degrees C per minute for either unfertilized or fertilized eggs.
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PMID:[Effects of cryoprotectants and sucrose dilution on the survival of unfertilized and fertilized mouse eggs after freezing and thawing]. 337 73

One hundred sixty-two embryos were collected from superovulated crossbred beef cattle 7 to 8 d after the onset of estrus. Embryos were frozen in modified Dulbecco's phosphate-buffered saline supplemented with 20% heat-inactivated fetal calf serum (PBS + FCS) and dimethylsulfoxide (DMSO), which was added in three steps to a final concentration of 1.5 M. Embryos were placed in .25 ml of 1.5 M DMSO in PBS + FCS in 1-ml glass ampules and cooled at 1.0 C/min from ambient temperature to -7 C, seeded and then cooled at .3 C/min to -19, -26, -33, -38, -43, -50 or -57 C before immersion (plunging) in liquid nitrogen. Ampules were thawed in 25 C water, and DMSO was removed in six steps at .25 M increments. 10 min/step. After removal of DMSO, embryos were cultured 24 h in PBS + FCS and then fixed and stained. Just after thawing, embryos for which slow cooling was terminated at -50 C were of lower (P less than .05) morphological quality than other groups. After removal of cryoprotectant, embryos from both the -19 and -50 C treatments had deteriorated more (P less than .05) than had embryos from other treatments. After 24-h culture, embryos slow-cooled to -19, -26 and -50 C had a lower rate of survival (P less than .05) than did embryos from -33, -38, -43 and -57 C temperatures. Embryos slow-cooled to -33, -38 and -43 C showed a higher percentage of healthy nuclei than did embryos slow-cooled to -19, -26 and -50 C.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Effect of slow cooling end point temperature on survival of frozen bovine embryos. 404 43

Anterior pituitaries from primiparous lactating C3H/He mice cultured in the medium containing 0.1% dimethyl sulfoxide (DMSO) for 48 hours and pituitaries from lactating mice given subcutaneous injections of 0.05 ml DMSO twice daily for two days and once on the morning of the third day were used in the studies of the in vitro and the in vivo effects of DMSO, respectively. Phosphate buffer saline was used in the control. Synthesis and release of growth hormone and prolactin were estimated by the incorporation of [14C]leucine into each hormone during three hours' incubation of the pituitaries pre-exposed to DMSO or PBS. The values in the medium represented released hormone and sum of the values in the medium and the pituitary represented the synthesized hormone. DMSO stimulated synthesis of GH and synthesis and release of prolactin in vitro. Meanwhile, in the vivo study, synthesis of GH and prolactin were lower in the DMSO-injected mice than in the control. The results suggest that the effects of DMSO on the pituitary secretion of GH and prolactin are adverse in vitro and in vivo. In vivo exposure of pituitary to DMSO resulted in the suppression of lactation.
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PMID:The in vitro and in vivo effects of dimethyl sulfoxide on the pituitary secretion of growth hormone and prolactin in mice. 619 45

Mouse morulae were frozen rapidly to -196 degrees C in the presence of 1.0-2.5 M-DMSO by a 3-step procedure; the samples were seeded at -4 to -8 degrees C, held at -20 degrees C in an ethanol bath for 10 min, suspended over liquid nitrogen at approximately -100 degrees C for 10 min and then plunged directly into liquid nitrogen at -196 degrees C. The cooling rate between -20 and -75 degrees C was approximately 17 degrees C/min. In all concentrations of DMSO significantly higher proportions of embryos developed to fully expanded blastocysts after 48 h in culture after rapid thawing (360 degrees C/min) than after slow thawing (25 degrees C/min). The highest survival rates were obtained for the embryos frozen rapidly in the presence of 1.5 and 2.0 M-DMSO (36 and 53% respectively). Various methods for removal of DMSO (2.0 M) were tested with the 3-step freezing and rapid thawing procedures. The best results for development to fully expanded blastocysts were obtained with PBS + 2.0 M DMSO + 0.5 M-sucrose (2 min) followed by PBS + 0.5 M-sucrose (2 min) at room temperature (82%) and with stepwise dilution in PBS at 30 degrees C (70%). When 26 embryos developed to blastocysts in culture after rapid freezing and thawing were transferred into 2 recipients, 11 newborn young (42%) were obtained.
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PMID:Survival of mouse embryos frozen and thawed rapidly. 740 Oct 45


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