共查询到20条相似文献,搜索用时 31 毫秒
1.
Seki S Kouya T Hara T Valdez DM Jin B Kasai M Edashige K 《The Journal of reproduction and development》2007,53(3):597-604
Movement of water and cryoprotectants through the plasma membrane needs to be accelerated for successful cryopreservation of zebrafish oocytes/embryos, which are much larger than their mammalian counterparts. Aquaporin-3 is a water/solute channel that can transport not only water but also various cryoprotectants. In this study, we attempted to increase the permeability of immature zebrafish oocytes at stage III to water and cryoprotectants by exogenous expression of rat aquaporin-3. Immature zebrafish oocytes were injected with rat aquaporin-3 cRNA and cultured for 5-12 h. Permeability to water and cryoprotectants was then determined based on changes in the volumes of the oocytes in a hypertonic sucrose solution and various cryoprotectant solutions at 25 C. The permeability to water of the aquaporin-3 cRNA-injected oocytes was three times higher than that of intact and water-injected oocytes. The permeability of the aquaporin-3 cRNA-injected oocytes to ethylene glycol, glycerol, propylene glycol, and DMSO was also 2-4 times higher than that of intact oocytes. Thus, the permeability of immature zebrafish oocytes to water and cryoprotectants was enhanced by exogenous expression of aquaporin-3. Cryopreservation of teleost oocytes may be realized through a further increase in permeability. 相似文献
2.
Pedro PB Yokoyama E Zhu SE Yoshida N Valdez DM Tanaka M Edashige K Kasai M 《The Journal of reproduction and development》2005,51(2):235-246
To assess the permeability of mouse oocytes and embryos, matured oocytes and embryos at various stages of development were placed in five cryoprotectant solutions at 25 C for 25 min. From the cross-sectional areas of the oocytes/embryos, the relative change in volume was analyzed. In oocytes, shrinkage was least extensive and recovery was quickest in the propylene glycol solution, showing that propylene glycol permeates the oocytes most rapidly. Dimethyl sulfoxide, acetamide, and ethylene glycol permeated the oocytes slightly more slowly than propylene glycol. The oocytes in glycerol shrunk extensively and then expanded marginally, indicating slow permeation. The volume changes of 1-cell and 2-cell embryos were similar to those of oocytes, showing little change in permeability. In 8-cell embryos, the volume recovered much faster than in the earlier stages especially in glycerol and acetamide. In morulae, the volume recovery was much faster in glycerol and in ethylene glycol; in ethylene glycol, the extent of shrinkage was small and the recovery was fast, indicating an extremely rapid permeation. Although the permeability of oocytes/embryos generally increased as embryo development proceeded, the degree of increase varied greatly among the cryoprotectants. Interestingly, the volume change in propylene glycol was virtually unaffected by the stage of development. Such information will be valuable for determining a suitable protocol for the cryopreservation of oocytes/embryos at different stages of development. 相似文献
3.
A.I. Moore MS PhD E.L. Squires PhD J.E. Bruemmer MS PhD J.K. Graham PhD 《Journal of Equine Veterinary Science》2006,26(5):215-218
Cryosurvival of cells is reduced if the cooling rate used is suboptimal. If cells cool too rapidly, intracellular water will freeze, causing intracellular ice crystals. However, if spermatozoa are cooled too slowly, excessive cellular dehydration occurs, causing irreversible damage to cellular compartments. In addition, cryoprotectants are added to the freezing diluent to protect cells from damage during cryopreservation. This study was conducted to determine the optimal cooling rate for stallion spermatozoa frozen in the presence of three different cryoprotectants. Spermatozoa were frozen in a skim milk, egg yolk diluent containing 4% glycerol, and ethylene glycol or dimethyl formamide at 10 different cooling rates ranging from 5°C/min to 50°C/min. The percentage of viable spermatozoa was higher for spermatozoa cooled at 10°C/min than at 50°C/min (P < .05). Spermatozoa frozen using glycerol as the cryoprotectant had higher percentages of motile and progressively motile spermatozoa compared with spermatozoa frozen using the other two cryoprotectants (P < .05). In conclusion, the cryosurvival of stallion spermatozoa is similar when cooling rates of 5°C/min to 45°C/min are used, and when 4% cryoprotectant is used, glycerol is a more effective cryoprotectant than ethylene glycol or dimethyl formamide. 相似文献
4.
Cryopreservation of Sheep Primordial Follicles 总被引:3,自引:0,他引:3
CA Amorim D Rondina CM Lucci A Giorgetti JR de Figueiredo PBD Gonçalves 《Reproduction in domestic animals》2007,42(1):53-57
The aim of this study was to evaluate the efficiency of 1 M dimethylsulphoxide (DMSO), ethylene glycol (EG), propylene glycol (PROH) and glycerol (GLY) to cryopreserve primordial follicles. The first evaluation was performed soon after cryopreservation and the second evaluation after 4 days of in vitro culture, using the cryoprotectants that allowed the higher results (higher follicular survival rate) after cryopreservation. The results after follicular isolation (control) and cryopreservation using 1 M DMSO, EG, PROH and GLY showed that the mean number (+/- SEM) of live follicles per millilitre was 3204 (100%) +/- 319.27, 2798 (87%) +/- 239.14, 2492 (78%) +/- 345.8, 448 (14%) +/- 46.3 and 208 (7%) +/- 75.26, respectively. Higher follicular survival was reported when DMSO and EG were used. Control follicles and follicles cryopreserved with these two cryoprotectants were cultured and the percentage of follicular survival was 55% (control), 42% (EG) and 34% (DMSO). Similar results were found between control and follicles cryopreserved with EG. In conclusion, 1 M EG is the most effective cryoprotectant to preserve primordial follicles isolated from ovaries of sheep. 相似文献
5.
Comparison of Ethylene Glycol and Propylene Glycol for the Vitrification of
Immature Porcine Oocytes
Tamás SOMFAI Michiko NAKAI Fuminori TANIHARA Junko NOGUCHI Hiroyuki KANEKO Naomi KASHIWAZAKI István EGERSZEGI Takashi NAGAI Kazuhiro KIKUCHI 《The Journal of reproduction and development》2013,59(4):378-384
Our aim was to optimize a cryoprotectant treatment for vitrification of immature porcine
cumulus-oocyte complexes (COCs). Immature COCs were vitrified either in 35% ethylene
glycol (EG), 35% propylene glycol (PG) or a combination of 17.5% EG and 17.5% PG. After
warming, the COCs were in vitro matured (IVM), and surviving oocytes were
in vitro fertilized (IVF) and cultured. The mean survival rate of
vitrified oocytes in 35% PG (73.9%) was higher (P<0.05) than that in 35% EG (27.8%).
Oocyte maturation rates did not differ among vitrified and non-vitrified control groups.
Blastocyst formation in the vitrified EG group (10.8%) was higher (P<0.05) than that in
the vitrified PG group (2.0%) but was lower than that in the control group (25.0%).
Treatment of oocytes with 35% of each cryoprotectant without vitrification revealed a
higher toxicity of PG on subsequent blastocyst development compared with EG. The
combination of EG and PG resulted in 42.6% survival after vitrification. The maturation
and fertilization rates of the surviving oocytes were similar in the vitrified, control
and toxicity control (TC; treated with EG+PG combination without cooling) groups.
Blastocyst development in the vitrified group was lower (P<0.05) than that in the
control and TC groups, which in turn had similar development rates (10.7%, 18.1% and
23.3%, respectively). In conclusion, 35% PG enabled a higher oocyte survival rate after
vitrification compared with 35% EG. However, PG was greatly toxic to oocytes. The
combination of 17.5% EG and 17.5% PG yielded reasonable survival rates without toxic
effects on embryo development. 相似文献
6.
Jiménez-Trigos E Naturil-Alfonso C Vicente JS Marco-Jiménez F 《Reproduction in domestic animals = Zuchthygiene》2012,47(3):472-478
Although much progress has been made in oocyte cryopreservation since 1971, live offspring have only been obtained in a few species and in rabbits. The aim of our study was to evaluate the effect of vitrification and slow freezing on the meiotic spindle, cortical granule (CG) distribution and their developmental competence. Oocytes were vitrified in 16.84% ethylene glycol, 12.86% formamide, 22.3% dimethyl sulphoxide, 7% PVP and 1% of synthetic ice blockers using Cryotop as device or slow freezing in 1.5 m PROH and 0.2 m sucrose in 0.25 ml sterile French mini straws. Meiotic spindle and CG distribution were assessed using a confocal laser-scanning microscope. To determine oocyte competence, in vitro development of oocytes from each cryopreservation procedure was assessed using parthenogenesis activation. Our data showed that oocytes were significantly affected by both cryopreservation procedures. In particular, meiotic spindle organization was dramatically altered after cryopreservation. Oocytes with peripheral CG distribution have a better chance of survival in cryopreservation after slow-freezing procedures compared to vitrification. In addition, slow freezing of oocytes led to higher cleavage and blastocyst rates compared to vitrification. Our data showed that, in rabbits, structural alterations are more evident in vitrified oocytes than in slow-frozen oocytes, probably as a consequence of sensitivity to high levels of cryoprotectants. Slow-freezing method is currently the recommended option for rabbit oocyte cryopreservation. 相似文献
7.
Effects of five cryoprotectants on proliferation and differentiation‐related gene expression of frozen‐thawed bovine calf testicular tissue 下载免费PDF全文
Hao Li Yi‐Lin Bian Nicola Schreurs Xiao‐Gang Zhang Sayed Haidar Abbas Raza Qian Fang Li‐Qiang Wang Jian‐Hong Hu 《Reproduction in domestic animals》2018,53(5):1211-1218
The cryopreservation of testicular tissue is a potential method for preserving male fertility. However, the effect of cryopreservation on bovine calf testicular tissue is scarce. This study investigated the effect of different cryoprotectants on bovine calf testicular tissue at the molecular level. Testicular tissue from ten immature bovine calves (6 months) was collected after slaughter and cryopreserved in an extender containing different concentrations of the following five cryopreservation solutions (CP): bovine serum albumin (BSA) with 5% dimethyl sulfoxide (DMSO), trehalose with 5% DMSO, DMSO and glycerol and ethylene glycol (EG). After 7‐day cryopreservation, the expression levels of three spermatogonial stem cell (SSC)‐related genes, octamer‐4 (OCT4), KIT ligand (MGF/SCF) and kit oncogene (C‐KIT), were investigated by quantitative PCR (qPCR). The cell viability was highest for the tissues preserved with 30 mg/ml BSA (77.82% ± 1.22) and 40 mg/ml trehalose (74.23% ± 1.16) compared with other groups (p < 0.05), and the level of expression of the three genes was highest with 30 mg/ml BSA (p < 0.05). Compared with other CPs, the 30 mg/ml BSA and 40 mg/ml trehalose have the better cryopreserve protection. The 30 mg/ml BSA is the most viable media for the cryopreservation of testicular tissue from cattle. 相似文献
8.
A. Massip 《Reproduction in domestic animals》2001,36(2):49-55
This review contains two parts. The first part is devoted to the significant steps in cryopreservation of mammalian embryos with emphasis on cattle and sheep that serve as models of reference. These steps are: (1) shortening of cooling and warming processes; (2) addition and dilution of cryoprotectant in one step; (3) introduction of plastic straw as a freezing and dilution container; (4) the choice of ethylene glycol as the quite universal cryoprotectant because of its low toxicity and high permeability; (5) vitrification, a cryopreservation method which enable passage from the liquid to the solid state by extreme elevation of viscosity due to high concentration of cryoprotectants and very rapid cooling. There are several vitrification solutions which contain dimethyl sulphoxide, glycerol, ethylene glycol, or a mixture of them, as basic cryoprotectants. The second part considers some factors affecting the efficiency of cryopreservation concerning (i) the origin of embryos and (ii) the stage of development and species. The origin of embryos (in vivo versus in vitro): in vitro embryos show a chilling and freezing sensitivity associated with their lipid content which can be modified by the culture conditions. Both conventional freezing and vitrification have been used and it seems that vitrification is more adapted to in vitro embryos when some modifications of initial protocols are carried out, particularly the rate of cooling. Thus considerable progress has been achieved by using the open pulled straw method of Vajta which enables the use of a minimum volume of freezing medium (0.5 μl) and a very high cooling rate that permits rapid traversal of the damaging temperature zone, corresponding to chilling sensitivity. The stage of development and species: not only are there differences between species at the same stage of development but in the same species all stages of development do not survive equally under the same freezing protocol. In cattle for example, oocytes and early stages of development in vivo or in vitro do not survive whereas compacted morulae and blastocysts survive very well. In the pig hatched blastocysts survive better than the other stages. Horse embryos have special characteristics that pose problems for successful freezing. In conclusion, a lot of work remains to be done to define fundamental characteristics of embryos of certain species (pig, horse) and of embryos of some stages or of oocytes. 相似文献
9.
Jiang BIAN Tao LI Chenhui DING Weijie XIN Bo ZHU Canquan ZHOU 《The Journal of reproduction and development》2013,59(3):288-295
To completely avoid ice crystal formation and thus get a higher survival rate,
vitrification methods have been commonly used for cryopreservation of oocytes and embryos.
However, currently used vitrification methods for oocytes and embryos are not suitable for
the cryopreservation of preantral follicles (PFs). In the present study, stainless steel
mesh was fabricated into mini mesh cups to vitrify isolated PFs. Moreover, isolated
follicles were encapsulated and then subjected to vitreous cryopreservation to facilitate
in vitro culture/maturation of follicles after warming. The results
showed that the percentages of viable follicles did not differ significantly between the
vitrification group and fresh group soon after warming (81.25% vs.
85.29%, P>0.05) and after a 7-day culture period (77.78% vs. 83.33%,
P>0.05). No difference in mean follicular diameter was observed between cryopreserved
and fresh follicles when cultured in vitro. Transmission electron
microscopic analysis revealed that vitreous cryopreservation could maintain the
ultrastructure of follicles in alginate beads. In conclusion, the present vitrification
method could efficiently cryopreserve isolated human ovarian follicles encapsulated by
calcium alginate, which could be put into immediate use (in vitro
culture/ maturation) after warming. However, more follicles and some detailed biochemical
analyses are required to further investigate the effects of vitrification on the long-term
growth of human encapsulated PFs. 相似文献
10.
Kohaya N Fujiwara K Ito J Kashiwazaki N 《The Journal of reproduction and development》2011,57(6):675-680
Unfertilized oocytes are one of the most desired germ cell stages for cryopreservation because these cryopreserved oocytes can be used for assisted reproductive technologies, including in vitro fertilization (IVF) and intracytoplasmic sperm injection. However, in general, the fertility and developmental ability of cryopreserved oocytes are still low. The aim of the present study was to improve vitrification of mouse oocytes. First, the effects of calcium and cryoprotectants, dimethyl sulfoxide and ethylene glycol (EG), in vitrification medium on survival and developmental ability of vitrified oocytes were evaluated. Oocytes were vitrified by a minimal volume cooling procedure using different cryoprotectants. Most of the vitrified oocytes were morphologically normal after warming, but their fertility and development were low independently of calcium and cryoprotectants. Second, the effect of cumulus cells on ability of oocytes to be fertilized and develop in vitro was examined. The fertility and developmental ability of denuded oocytes (DOs) after IVF were reduced compared with cumulus-oocyte complexes (COCs) both in fresh and cryopreserved groups. Vitrified COCs showed significantly (P<0.05) higher fertility and ability to develop to the 2-cell and blastocyst stages than those of vitrified DOs with cumulus cells and vitrified DOs alone. The vitrified COCs developed to term at a high success rate equivalent to the rate obtained with IVF using fresh COCs. Taken together, the current results clearly demonstrate that, in the presence of surrounding cumulus cells, matured mouse oocytes vitrified using calcium-free media and EG retain their developmental competence. These findings will contribute to improve oocyte vitrification in not only experimental animals but also clinical application for human infertility. 相似文献
11.
Jin B Yamasaki C Yamada N Seki S Valdez DM Kasai M Edashige K 《The Journal of reproduction and development》2008,54(4):265-269
To improve the cryopreservation protocol for mouse sperm, we attempted to estimate the type and extent of cryoinjury at various steps of the process. First, we demonstrated that mouse sperm are sensitive to chilling at -15 C and that the sensitivity is dependent on the length of exposure. To estimate cryoinjuries, sperm suspensions were ice-seeded at -5 or -15 C, frozen with liquid nitrogen (LN(2)) gas and then frozen in LN(2). In one experiment, sperm seeded at -5 C were cooled slowly to -15 C before deep freezing. At various steps of the cryopreservation process, the sperm were warmed and their viability was assessed based on motility and the integrities of the plasma membrane and acrosome. The motility of frozen-thawed sperm was higher on seeding at -5 C (28%) than at -15 C (9%). The motility did not decrease when the sample was transferred from LN(2) gas to LN(2). To estimate cryoinjury of sperm, we presumed the viability of frozen sperm to be decreased by chilling, hypertonic stress and formation of intracellular ice. When the sperm suspension was cooled and seeded at -5 C, the motility decreased by 25% due to hypertonic stress. When the sperm were cooled in LN(2) gas, the motility decreased by 17% with the formation of intracellular ice. When the sperm were cooled to -15 C, the motility decreased by 51% from chilling. After seeding, the motility decreased by 18% due to formation of intracellular ice and by 7% due to hypertonic stress. Considering the results, it would be preferable to seed samples at a higher temperature to prevent intracellular ice from forming and to cool seeded samples rapidly enough to minimize chilling injury and hypertonic stress, but not too rapidly to allow intracellular ice to form. 相似文献
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13.
KGhM Mahmoud TH Scholkamy YF Ahmed GE Seidel Jr MF Nawito 《Reproduction in domestic animals》2010,45(4):565-571
This study was designed to evaluate effects of different combinations of cryoprotectants on the ability of vitrified immature buffalo oocytes to undergo in vitro maturation. Straw and open‐pulled straw (OPS) methods for vitrification of oocytes at the germinal vesicle stage also were compared. The immature oocytes were harvested from ovaries of slaughtered animals and were divided into three groups: (i) untreated (control); (ii) exposed to cryoprotectant agents (CPAs); or (iii) cryopreserved by straw and OPS vitrification methods. The vitrification solution (VS) consisted of 6 m ethylene glycol (EG) as the standard, control vitrification treatment, and this was compared with 3 m EG + 3 m dimethyl sulfoxide (DMSO), 3 m EG + 3 m glycerol, and 3 m DMSO + 3 m glycerol. Cryoprotectants were added in two steps, with the first step concentration half that of the second (and final) step concentration. After warming, oocyte samples were matured by standard methods and then fixed and stained for nuclear evaluation. Rates of MII oocytes exposed to CPAs without vitrification were lower (54.3 ± 1.9% in EG, 47.5 ± 3.4% in EG + DMSO, 36.8 ± 1.2% in EG + glycerol and 29.9 ± 1.0% in DMSO + glycerol; p < 0.05) than for the control group (79.8 ± 1.3%). For all treatments in each vitrification experiment, results were nearly identical for straws and OPS, so all results presented are the average of these two containers. The percentages of oocytes reaching telophase‐I or metaphase‐II stages were lower in oocytes cryopreserved using all treatments when compared with control. However, among the vitrified oocytes, the highest maturation rate was seen in oocytes vitrified in EG + DMSO (41.5 ± 0.6%). Oocytes cryopreserved in all groups with glycerol had an overall low maturation rate 19.0 ± 0.6% for EG + glycerol and 17.0 ± 1.1% for DMSO + glycerol. We conclude that the function of oocytes was severely affected by both vitrification and exposure to cryoprotectants without vitrification; the best combination of cryoprotectants was EG + DMSO for vitrification of immature buffalo oocytes using either straw or OPS methods. 相似文献
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Yamaji Y Seki S Matsukawa K Koshimoto C Kasai M Edashige K 《The Journal of reproduction and development》2011,57(3):403-408
Previously, we showed that the exogenous expression of aquaporin 3 (AQP3), an aquaglyceroporin, improved the tolerance of mouse oocytes to vitrification with a glycerol-based solution. In the present study, we examined conditions suitable for the expression of AQP3 and the ability of vitrified oocytes to develop in vitro and in vivo after fertilization. After only partial remove of cumulus cells, immature mouse oocytes (germinal vesicle stage) were injected with 5, 10 or 20 pg of AQP3 cRNA and cultured for 12 h for maturation. When matured oocytes were vitrified with a glycerol-based solution, 57-61% survived regardless of the amount of cRNA injected (5-20 pg). By contrast, no oocytes injected with water (control) survived. When the zona pellucida was removed from the vitrified oocytes and the oocytes were then fertilized in vitro and cultured, the proportions that were fertilized and developed into blastocysts were higher when the amount of cRNA injected was 5 pg than 10-20 pg. When 16 blastocysts were transferred to a pseudopregnant mouse, 5 developed to term, demonstrating that oocytes vitrified after injection of AQP3 cRNA retained the ability to develop to term. The water-permeability of cRNA-injected oocytes was higher than that of control oocytes from the maturing stage to the 1-cell zygote stage, whereas glycerol-permeability was higher only at metaphase II. This indicates that AQP3 was expressed for a relatively short period of time. These results suggest that the transient expression of water/cryoprotectant channels is effective for cryopreserving cells that have low membrane-permeability, such as mammalian oocytes. 相似文献
16.
Hochi S 《The Journal of reproduction and development》2003,49(1):13-21
Factors affecting sensitivity of preimplantation embryos and follicular oocytes to cryopreservation were analyzed in the equine and bovine species. (1) Survival of equine blastocysts after two-step freezing in the presence of glycerol as the cryoprotective agent (CPA) was influenced by development of the embryonic capsule. The use of ethylene glycol (EG) with sucrose as CPAs improved the post-thaw survival of blastocysts and made it possible to transfer the embryos into recipient mares without removing the CPAs. In addition, early blastocysts cryopreserved by vitrification could develop both in vitro and in vivo when the embryos were exposed to vitrification solution in a stepwise manner. The vitrification procedure was also applied to the relatively large expanded blastocysts. (2) Bovine embryos produced in vitro have been considered to be highly sensitive to the process of cryopreservation. To solve this problem, Day-7 blastocysts produced in a serum-free system were cooled at 0.3 C/min rather than 0.6 C/min before being plunged into liquid nitrogen, resulting in no loss of the post-thaw viability. The supplementation of LAA in IVM/IVF media or IVC medium was effective in producing pronuclear-stage zygotes or morula-stage embryos relatively tolerable to freezing, respectively. (3) Transmission electron microscopic observation of immature equine oocytes showed that cellular injury occurred near the sites of gap-junctions between cumulus cells and the oocyte. In cattle, higher fertilization rates of oocytes were obtained when the oocytes were subjected to cryopreservation at an intermediate stage during IVM (GVBD for freezing, Met-I for vitrification). Vitrification of bovine Met-II oocytes in open-pulled glass capillaries, characterized by an ultra-rapid cooling rate (3,000-5,000 C/min), was found to avoid any harmful influence of vitrification and warming. 相似文献
17.
研究了麦管和OPS管法冷冻以及OPS法中保护剂种类对牛卵母细胞体外成熟及孤雌胚发育的影响。结果发现,OPS管冷冻牛卵母细胞形态正常率、卵裂率、囊胚率极显著高于麦管法(P<0.05)。在OPS法中,应用两种保护剂冷冻后,卵母细胞形态正常率、卵裂率、囊胚率差异极显著(P<0.01);采取38℃与25℃温度平衡,冷冻后卵母细胞各发育指标差异不显著(P>0.05);采用4℃平衡,冷冻后的卵母细胞激活后没有出现囊胚,各发育指标极显著降低(P<0.01)。结果表明,OPS法可以有效地保护卵母细胞,保证其后孤雌激活;采用低温平衡会对孤雌发育的囊胚阶段有较大影响。 相似文献
18.
抗冻蛋白(antifreeze proteins,AFPs)是一类能抑制冰晶生长并提高机体抗冻功能的蛋白质。AFPs来源广泛,在动物、植物、细菌、真菌等不同生物体内都存在。目前,研究较多、有望被推广应用于畜牧生产实践的主要是具有热滞活性和抑制冰晶重结晶活性的鱼类AFPs。鱼类AFPs的种类主要包括AFPⅠ、AFPⅡ、AFPⅢ、AFPⅣ和抗冻糖蛋白(AFGP)。尽管不同鱼类AFPs都具有抗冻功能,但是它们的组成和结构差异很大,而且对应的基因在核苷酸序列上几乎没有相似性。鱼类AFPs抗冻机制主要包括吸附-抑制学说、刚体能量学说、包合物锚定学说、亲和相互作用偶联团聚学说等。在人类、鱼类、家畜和小鼠等精液的低温保存中,鱼类AFPs可通过维持细胞膜完整性、抑制冰晶生长及减少DNA损伤等保护精子;在卵母细胞、精原细胞、红细胞及动物胚胎的低温保存中,鱼类AFPs可通过阻止冰晶化及抗氧化功能等改善保存效果。作者主要概述了鱼类AFPs种类及其结构特征和作用机制,总结了鱼类AFPs在精液、卵母细胞、精原细胞、红细胞及胚胎低温保存中的应用进展,以期为鱼类AFPs作为候选抗冻剂应用于种质资源保护和畜牧业生产提供科学依据。 相似文献
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FC Varago VS Moutacas BC Carvalho RV Serapião F Vieira H Chiarini‐Garcia FZ Brandão LS Camargo M Henry MA Lagares 《Reproduction in domestic animals》2014,49(5):839-844
The aim of this work was to evaluate the efficiency of the cryoprotectants dimethylformamide and ethylene glycol for cryopreservation of ovine embryos using vitrification and conventional freezing. The recovered embryos were distributed randomly in three treatment groups: Gr. 1: conventional freezing (n = 44), Gr. 2: vitrification with ethylene glycol (n = 39) and Gr. 3: vitrification with dimethylformamide (n = 38). Quality of fresh embryos in control group as well as of frozen and vitrified embryos was examined by three methodologies: staining with propidium iodide and Hoechst 33258 and evaluation under fluorescent microscopy, evaluation of re‐expansion and hatching rates after culture, and determination of apoptotic index with TUNEL technique. It was established that re‐expansion rate in all treatment groups was similar. In the same time, hatching rates were higher in Gr. 1 (40.5%) and Gr. 2 (35.3%) in comparison with Gr. 3 (15.5%, p < 0.05). The number of dead cells in vitrified embryos of Gr. 2 and Gr. 3 was higher (42.6 ± 26.2 and 63.2 ± 34.65, respectively) in comparison with Gr. 1 (conventional freezing, 10.1 ± 8.5, p < 0.05). Embryos vitrified with dimethylformamide included the same quality of apoptotic cells that Gr. 1 (conventional freezing) and fresh embryos. In conclusion, the dimethylformamide and ethylene glycol used as cryoprotectant to vitrify ovine embryos, in the concentrations and exposition time tested in this work, were not as efficient as the conventional freezing for cryopreservation of ovine embryos Thus, the conventional freezing with ethylene glycol was the most efficient method to cryopreserve ovine embryos in comparison with vitrification. 相似文献