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1.
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
2.
Tatsuya NAKANO Mizuki KONO Kazuki SEGAWA Satoshi KUROSAKA Yoshiharu NAKAOKA Yoshiharu MORIMOTO Tasuku MITANI 《The Journal of reproduction and development》2021,67(2):123
Methylglyoxal (MG) is a precursor for the generation of endogenous advanced glycation end-products involved in various diseases, including infertility. The present study evaluated the motility and developmental competence after in vitro fertilization of mouse sperm which were exposed to MG in the capacitation medium for 1.5 h. Sperm motility was analyzed using an SQA-V automated sperm quality analyzer. Intracellular reactive oxygen species (ROS), membrane integrity, mitochondrial membrane potential, and DNA damage were assessed using flow cytometry. The matured oocytes were inseminated with MG-exposed sperm, and subsequently, the fertilization and embryonic development in vitro were evaluated in vitro. The exposure of sperm to MG did not considerably affect the swim-up of sperm but resulted in a deteriorated sperm motility in a concentration-dependent manner, which was associated with a decreased mitochondrial activity. However, these effects was not accompanied by obvious ROS accumulation or DNA damage. Furthermore, MG diminished the fertilization rate and developmental competence, even after normal fertilization. Collectively, a short-term exposure to MG during sperm capacitation had a critical impact on sperm motility and subsequent embryonic development after fertilization. Considering that sperm would remain in vivo for up to 3 days until fertilization, our findings suggest that sperm can be affected by MG in the female reproductive organs, which may be associated with infertility. 相似文献
3.
In vitro growth of immature oocytes provides opportunities to increase gametic resources and
to understand the mechanisms underlying oocyte development. Many studies on the in vitro
growth of oocytes have been reported thus far; however, only a few cases have been reported, which
demonstrated that oocytes can support full-term development after in vitro fertilization. Our
research group recently found that culture of mouse neonatal primordial follicles increased the birthrate;
however, the establishment of an in vitro system that can completely mimic follicle or oocyte
growth in vivo and control oogenesis remains an ongoing challenge. 相似文献
4.
Shun TAKEO Daichi SATO Koji KIMURA Yasunori MONJI Takehito KUWAYAMA Ryoka KAWAHARA-MIKI Hisataka IWATA 《The Journal of reproduction and development》2014,60(2):92-99
The aim of the present study was to address the effect of resveratrol-mediated
upregulation of sirtuin 1 (SIRT1) during oocyte maturation on mitochondrial function, the
developmental ability of oocytes and on mechanisms responsible for blockage of polyspermic
fertilization. Oocytes collected from slaughterhouse-derived ovaries were cultured in
TCM-199 medium supplemented with 10% FCS and 0 or 20 µM resveratrol (Res). We examined the
effect of Res on SIRT1 expression in in vitro-matured oocytes (Exp 1);
fertilization and developmental ability (Exp 2); mitochondrial DNA copy number (Mt
number), ATP content and mitochondrial membrane potential in matured oocytes (Exp 3); and
the time required for proteinase to dissolve the zona pellucida following in
vitro fertilization (as a marker of zona pellucida hardening), as well as on
the distribution of cortical granules before and after fertilization (Exp 4). In Exp 1,
the 20 µM Res treatment upregulated protein expression of SIRT1 in oocytes. In Exp 2, Res
treatment improved the ratio of normal fertilization and the total cell number of
blastocysts. In Exp 3, Res treatment significantly increased the ATP content in matured
oocytes. Additionally, Res increased the overall Mt number and mitochondrial membrane
potential, but the effect was donor-dependent. In Exp 4, Res-induced zona hardening
improved the distribution and exocytosis of cortical granules after in
vitro fertilization. In conclusion, Res improved the quality of oocytes by
improving mitochondrial quantity and quality. In addition, Res added to the maturation
medium enhanced SIRT1 protein expression in oocytes and improved fertilization via
reinforcement of the mechanisms responsible for blockage of polyspermic fertilization. 相似文献
5.
Ayumi HASEGAWA Keiji MOCHIDA Toshiko TOMISHIMA Kimiko INOUE Atsuo OGURA 《The Journal of reproduction and development》2014,60(3):187-193
Successful in vitro fertilization (IVF) in mice has been achieved using spermatozoa at concentrations
specifically optimized for the experimental conditions, such as species and source of spermatozoa. Although IVF in mice is
mostly performed using about 80–500 µl drops, it is expected that the number of spermatozoa used for insemination can be
reduced by decreasing the size of the IVF drops. The present study was undertaken to examine the extent to which the number
of spermatozoa used for IVF could be reduced by using small droplets (1 µl). We devised the experimental parameters using
frozen–thawed spermatozoa from C57BL/6 mice in anticipation of broader applications to other mouse facilities. We found that
as few as 5 spermatozoa per droplet could fertilize oocytes (1 or 3 oocytes per droplet), although the fertilization rates
were low (13–15%). Practical fertilization rates (> 40%) could be achieved with frozen-thawed C57BL/6J spermatozoa, which
are sensitive to cryopreservation, when 20 sperm per droplet were used to inseminate 3 oocytes. Even with spermatozoa from a
very poor quality suspension (10% motility), about 25% of oocytes were fertilized. Our calculations indicate that the number
of inseminated spermatozoa per oocyte can be reduced to 1/96–1/240 by this method. In two separate embryo transfer
experiments, 60% and 47%, respectively, of embryos developed to term. Our microdroplet IVF method may be particularly
advantageous when only a limited number of motile spermatozoa are available because of inadequate freezing-thawing or genetic
reasons. 相似文献
6.
Fan ZQ Li XW Liu Y Meng QG Wang YP Hou YP Zhou GB Zhu SE 《The Journal of reproduction and development》2008,54(2):107-112
This study was performed to investigate the effect of partial zona pellucida incision by piezo micromanipulation (ZIP) on the in vitro fertilizing ability of stored mouse spermatozoa. The storage conditions were optimized by storing the mouse epididymides at 4 C in mineral oil or in the mouse body for up to 4 days after death, and the retrieved spermatozoa were used to fertilize fresh oocytes. No significant difference was observed in fertilization rates between the treatments when epididymides were stored for up to 2 days, but the fertilization rates in mineral oil were higher (P<0.05) than those in the mouse body at 3 (41.4 vs. 16.2%) and 4 days (26.0 vs. 15.8%). Spermatozoa retrieved from epididymides stored in mineral oil were then used to fertilize fresh and vitrified oocytes with or without ZIP treatment. The fertilization rates of the ZIP fresh oocytes were higher than those of the zona-intact oocytes at each time point (1 to 4 days). After ZIP, the fertilization rates of spermatozoa stored for 1 and 2 days (91.2 and 86.6%, respectively) were similar (P>0.05) to that of fresh spermatozoa (91.9%). In regard to vitrified oocytes, the fertilization rates of zona-intact and ZIP oocytes using fresh spermatozoa were 46.7 and 84.7%, while the fertilization rates of vitrified ZIP oocytes using spermatozoa stored for 1 to 4 days ranged from 49.3 to 79.6%. When 2-cell embryos derived from ZIP fresh and vitrified oocytes inseminated with 2 day-stored spermatozoa were transferred into recipient females, 47.9 and 15.0% of the embryos developed to term, respectively. These results indicate that storing mouse epididymides at 4 C in mineral oil is more suitable than storage in the mouse body and that the ZIP technique improves the in vitro fertilizing ability of stored mouse spermatozoa in fresh oocytes and significantly increases the fertilization rate of vitrified oocytes with fresh spermatozoa. 相似文献
7.
Sadamasa ISHIKAWA Kou HIRAGA Yuuki HIRADATE Kentaro TANEMURA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(6):725-728
Acetamiprid (ACE) and imidacroprid (IMI) are known neonicotinoid insecticides with strong
affinities for the insect-selective nicotinic acetylcholine receptor. These provide insect
control by hyperstimulating insect nerves and are used for agricultural pest management.
However, it has also been reported that ACE and IMI affect mammalian reproductive
function. We determined the effects of ACE and IMI on the in vitro
maturation of porcine oocytes. Significant decreases in nuclear maturation rates were
observed in the ACE or IMI-exposed groups. Also, in matured oocytes from the ACE or
IMI-exposed groups, irregular chromosomes were observed. Our results suggest that ACE and
IMI exposure was detrimental to porcine oocytes and the extent of the effects depends on
the concentration of exposure. 相似文献
8.
Kou HIRAGA Yumi HOSHINO Kentaro TANEMURA Eimei SATO 《The Journal of reproduction and development》2013,59(4):405-408
Localization patterns of lipid droplets in the cytoplasm of porcine oocytes were
evaluated as a novel marker for in vitro maturation (IVM) of oocytes with
high developmental competence. Porcine oocytes were cultured in TCM-199, which is a
complete synthetic medium, for 44 h at 38.5 C. Localization patterns were divided into 2
classes: lipid droplets localized uniformly in the whole cytoplasm (class I) and those
that were centrally located (class II). After IVM in TCM-199, 60% of matured oocytes
exhibited the class II pattern. To investigate the relation between the distribution of
lipid droplets and the developmental rate of the oocyte, the developmental rates of class
I and class II oocytes were compared after in vitro fertilization (IVF).
Class II oocytes showed a significantly higher rate of blastocyst development than class I
oocytes. These results suggest that porcine oocytes with high developmental competence can
be selected based on the localization patterns of lipid droplets. 相似文献
9.
Naomi NAKAGATA Toru TAKEO Kiyoko FUKUMOTO Yukie HARUGUCHI Tomoko KONDO Yumi TAKESHITA Yuko NAKAMUTA Tomoko UMENO Shuuji TSUCHIYAMA 《The Journal of reproduction and development》2014,60(2):168-171
Sperm cryopreservation has been widely adopted for maintenance of the genetically
engineered mouse (GEM). The cryopreserved sperm are being exchanged among many institutes
worldwide. However, the recipients are not always able to obtain high fertilization rates
with the frozen sperm shipped from senders. In this study, we cryopreserved mouse sperm
via various methods and performed in vitro
fertilization (IVF) in which the combination of methyl-beta-cyclodextrin for sperm
preincubation and reduced glutathione for insemination was used (the MBCD-GSH IVF). In
addition, frozen sperm sent from the Jackson Laboratory (USA) were thawed and used for IVF
in the same manner. The fertilization rates of both the sperm cryopreserved
via the methods applied in some countries and the cryopreserved GEM
sperm improved when used with the MBCD-GSH IVF method. Therefore, we strongly believe that
the MBCD-GSH IVF method brings about relatively high fertilization rates with any strain
of frozen mouse sperm. 相似文献
10.
Studies were conducted to examine the possibility of preserving slaughterhouse‐derived buffalo ovaries at 4°C for 0 (control), 12 and 24 h to maintain the developmental competence of the oocytes (experiment 1), to assess the effect of incubation temperature during oocyte maturation on rates of in vitro maturation (IVM) and in vitro fertilization (IVF) of buffalo oocytes and embryo development (experiment 2), and to examine the effect of storage at 25°C for 0 (control), 4 and 8 h of frozen–thawed buffalo sperm and BO and H‐TALP as sperm processing and fertilization media on cleavage and embryo development in vitro of buffalo oocytes (experiment 3) in order to optimize the IVF technology in buffalo. Results suggested that storage of ovaries at 4°C for 12 or 24 h significantly (p < 0.05) reduced the developmental potential of oocytes. Incubation temperatures during the IVM influenced the fertilization rate but had no significant effect on maturation and subsequent embryo development. The incubation temperature of 38.5°C during IVM was found to be optimum for embryo production in vitro. Storage of frozen–thawed sperm at 25°C for 8 h significantly (p < 0.05) decreased its ability to cleave the oocytes. Sperm processed in BO medium had significantly (p < 0.05) higher ability to cleave the oocytes than the H‐TALP medium. 相似文献
11.
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
12.
Oocyte and Embryo Quality: Effect of Origin, Culture Conditions and Gene Expression Patterns 总被引:3,自引:0,他引:3
P Lonergan D Rizos A Gutierrez-Adan T Fair MP Boland 《Reproduction in domestic animals》2003,38(4):259-267
In general, the majority of immature bovine oocytes fail to develop to the blastocyst stage following maturation, fertilization and culture in vitro. The evidence suggests that while culture conditions during in vitro embryo production can impact on the developmental potential of the early embryo, the intrinsic quality of the oocyte is the key factor determining the proportion of oocytes developing to the blastocyst stage. In addition, evidence suggests that the period of post‐fertilization embryo culture is the most critical in determining blastocyst quality. This paper reviews the current literature, with emphasis on the bovine model, demonstrating evidence for an effect of oocyte origin and/or in vitro maturation conditions on the developmental capacity and gene expression patterns in the oocyte. Furthermore, the well‐documented effects of post‐fertilization culture environment on embryo gene expression and quality are highlighted. 相似文献
13.
Developmental competence of in vitro matured ovine oocytes vitrified in solutions with different concentrations of trehalose
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Batool Sanaei Bahar Movaghar Mojtaba Rezazadeh Valojerdi Bita Ebrahimi Masood Bazrgar Mehdi Hajian Mohammad H. Nasr‐Esfahani 《Reproduction in domestic animals》2018,53(5):1159-1167
This study aimed to determine the optimum concentration of trehalose in solutions used for vitrification of in vitro matured (IVM) ovine oocytes. IVM oocytes were randomly divided into four experimental (vitrified) and one control (fresh) groups. Experimental groups were treated with different concentrations (0.0, 0.25, 0.5 and 1.0 M) of trehalose. After warming, some viable oocytes were exposed to 0.25% pronase to test zona pellucida hardening, whereas the others were fertilized and cultured in vitro for 8 days to evaluate their developmental competence. Blastocysts quality was assessed by differential staining and TUNEL test. Survival and developmental rates of oocytes vitrified in the presence of 0.5 M trehalose were significantly higher than those of the other vitrified groups. Furthermore, there was a significant difference between fresh and vitrified groups in total blastocyst rate. Analysis of blastocysts quality also revealed a significant difference between the group treated with 0.5 M trehalose and other groups in terms of apoptotic index. Furthermore,zona pellucida digestion time period was longer in trehalose‐free (0.0 M) group compared to other groups. In conclusion, we found that IVM ovine oocytes vitrified in solutions containing 0.5 M trehalose are fertilization‐competent and are able to produce good‐quality blastocysts with an apoptotic index comparable to that of the fresh oocytes. Therefore, 0.5 M may be considered the optimum concentration of trehalose to be used in solutions prepared for vitrification of oocytes. 相似文献
14.
Keisuke KOYAMA Sung-Sik KANG Weiping HUANG Yojiro YANAGAWA Yoshiyuki TAKAHASHI Masashi NAGANO 《The Journal of reproduction and development》2014,60(2):136-142
The objective of this research was to clarify the aging-related changes in in
vitro-matured bovine oocytes. Firstly, we examined the fertilization and
embryonic development of bovine oocytes after 22 and 30–34 h of in vitro
maturation (IVM). The oocytes after 30–34 h of IVM (penetrated by sperm at around 40 h
after starting IVM) showed a lower developmental rate to blastocysts (P<0.01), although
normal fertilization rates were similar regardless of IVM duration. In the next
experiment, reactive oxygen species (ROS), mitochondrial activity and ATP content in
oocytes after 20, 30 and 40 h of IVM were examined. The lowest level of ROS was found in
the group subjected to 30 h of IVM. The mitochondrial activity and ATP content in the
group subjected to 40 h of IVM were higher than in the group subjected to 20 h of IVM
(P<0.01), and those in the group subjected to 30 h of IVM showed intermediate values.
Thereafter, the mitochondrial activities at 3 days after in vitro
fertilization in embryos derived from the oocytes subjected to 22 and 34 h of IVM were
evaluated. In the group subjected to 34 h of IVM, high-polarized mitochondria were
frequently observed at the periphery of blastomeres. The present results suggest that high
mitochondrial activity observed in oocytes after prolonged IVM culture and localization of
high-polarized mitochondria at the periphery of blastomeres during early embryonic
development may be associated with the low developmental competence in aged bovine
oocytes. 相似文献
15.
Seon-Hyang KIM Ming-Hui ZHAO Shuang LIANG Xiang-Shun CUI Nam-Hyung KIM 《The Journal of reproduction and development》2015,61(4):261-268
Cathepsin B, a lysosomal cysteine protease of the papain family, has recently been implicated in the quality and developmental competence of bovine preimplantation embryos. In this study, to determine whether inhibition of cathepsin B activity can improve porcine oocyte maturation and early embryo developmental competence, we supplemented in vitro maturation or embryo culture media with E-64, a cathepsin B inhibitor. Cathepsin B activity was high in poor quality germinal vesicle stage oocytes, but no differences in mRNA expression or protein localization were observed between good and poor quality oocytes, which were categorized based on morphology. Following treatment with 1 μM E-64, cathepsin B activity sharply decreased in 4-cell and blastocyst stage embryos. E-64 had no effect on cell number but significantly (P < 0.05) increased blastocyst formation and decreased the number of apoptotic cells in blastocysts. It also significantly (P < 0.05)
enhanced mitochondrial membrane potential in blastocysts, reducing the release of cytochrome c and resulting in decreased expression of Caspase-3 and Caspase-9. In conclusion, inhibition of cathepsin B activity in porcine parthenotes using 1 μM E-64 resulted in attenuation of apoptosis via a reduction in the release of cytochrome c from mitochondria. 相似文献
16.
A comparative study of Sephadex, glass wool and Percoll separation techniques on sperm quality and IVF results for cryopreserved bovine semen 总被引:1,自引:0,他引:1
Hae-Lee Lee Sue-Hee Kim Dong-Beom Ji Yong-Jun Kim 《Journal of veterinary science (Suw?n-si, Korea)》2009,10(3):249-255
The aim of this study was to compare the effects of spermatozoa separation techniques on sperm quality and in-vitro fertilization (IVF) results for cryopreserved bovine semen. Sephadex, glass wool and Percoll gradient separation techniques were used for sperm separation and sperm motility, morphology and membrane integrity were evaluated before and after separation. Also, cleavage and blastocyst developmental rate were investigated after IVF with sperm recovered by each separation technique. The motility of samples obtained by the three separation techniques were greater compared to the control samples (p < 0.05). The percentage of spermatozoa with intact plasma-membrane integrity, identified by 6-carboxyfluoresceindiacetate/propidium iodide fluorescent staining and the hypo-osmotic swelling test, was highest in the glass wool filtration samples (p < 0.05). The cleavage and blastocyst rate of total oocytes produced from glass wool filtration samples were also higher than the control and Sephadex filtration samples (p < 0.05), but were not significantly different from Percoll separation samples. However, a significantly greater number of cleaved embryos produced by glass wool filtration developed to blastocyst stage than those produced by Percoll separation (p < 0.05). These results indicate that spermatozoa with good quality can be achieved by these three separation techniques and can be used for bovine IVF. In particular, it suggests that glass wool filtration would be the most effective method of the three for improving sperm quality and embryo production for cryopreserved bovine spermatozoa. 相似文献
17.
Sung-Sik KANG Keisuke KOYAMA Weiping HUANG Yinghua YANG Yojiro YANAGAWA Yoshiyuki TAKAHASHI Masashi NAGANO 《The Journal of reproduction and development》2015,61(2):99-105
The present study aimed to establish an efficient system for bovine embryo production by in vitro fertilization (IVF) that can achieve stable normal fertilization and blastocyst developmental rates in any bull without optimization of the sperm concentration in IVF medium. We examined the effects of a PHE mixture (20 μM D-penicillamine, 10 μM hypotaurine and 1 μM epinephrine), theophylline (2.5 mM), and sperm concentration (1, 2 or 5 × 106 cells/ml) on fertilization and blastocyst developmental rates. High cleavage rates (78.3 to 92.4%) and blastocyst developmental rates (31.9 to 62.0%) at day 7 were obtained in the presence of PHE and theophylline in IVF medium with a sperm concentration of 2 × 106 cells/ml using sperm from 9 bulls. In addition, the synergistic effect of PHE and theophylline on normal fertilization (2 pronuclei) was clarified at 12 h after IVF with a sperm concentration of 1 × 106 cells/ml. Moreover, high
linearity, high flagellar beat cross frequency, and low amplitude of lateral head of motile sperm were found by computer-assisted sperm analysis. In conclusion, the combination of the PHE mixture and theophylline synergistically accelerates sperm motility and sperm penetration of bovine oocytes. Theophylline activates sperm motility with increasing intracellular cAMP. However, PHE prevents an excessive increase of cAMP and maintains sperm motility without hyperactivation. When the combination of PHE and theophylline is added to IVF medium at a sperm concentration of 2 × 106 cells/ml, we can achieve stable normal fertilization and blastocyst development in any bull. 相似文献
18.
Developmental competence and glutathione content of maternally heat-stressed mouse oocytes and zygotes 总被引:4,自引:0,他引:4
Takaya MATSUZUKA Manabu OZAWA Miho HIRABAYASHI Atsuko USHITANI Yukio KANAI 《Animal Science Journal》2004,75(2):117-124
The loss of developmental competence and the glutathione (GSH) content of maternally heat‐stressed mouse oocytes and zygotes were determined. In experiment 1, zygotes were collected from female mice that were heat‐stressed at 35°C for 10 h after hCG injection (oocyte maturation stage), or for 12 h on Day 1 of pregnancy (zygote stage), followed by in vitro culture. To minimize the effects of heat stress on the fertilization process, heat‐stressed oocytes that were fertilized in vitro were also included in this experiment. In experiment 2, heat‐stressed oocytes and zygotes were assayed for GSH content. The application of heat stress to the oocytes resulted in a significant decrease in the percentage of zygotes that developed to morulae or blastocysts, both for naturally fertilized oocytes (56.9% for heat‐stressed vs 85.4% for control) or in vitro‐fertilized oocytes (54.5%vs 73.6%). In the heat‐stressed zygotes, the disruption of embryonic development was more drastic (24.3%vs 90.3%), with the majority of zygotes being arrested at the two‐cell stage. In contrast, the GSH content decreased significantly in heat‐stressed zygotes, but not in heat‐stressed oocytes. These results demonstrate that the loss of developmental competence of early embryos is associated with a decrease in the GSH content of maternally heat‐stressed zygotes, but not of maternally heat‐stressed oocytes. 相似文献
19.
Budiyanto AGUNG Takeshige OTOI Dai-ichiro FUCHIMOTO Shoichiro SENBON Akira ONISHI Takashi NAGAI 《The Journal of reproduction and development》2013,59(2):103-106
This study was conducted to assess the fertilization and development of porcine oocytes
matured in a solo follicular fluid (pFF) using different in vitro culture
systems and insemination periods. Cumulus-oocyte complexes (COCs), follicular cells (FCs),
and pFF were collected from the follicles of ovaries. The pFF was used as a maturation
medium (MpFF) after supplementation with follicle stimulating hormone (FSH) and
antibiotics. The COCs were matured in a 15 ml test tube containing 3.5 ml of MpFF with FCs
(5.2 × 106 cells/ml; rotating culture system) or 2 ml of MpFF without FCs in a
35-mm petri dish (static culture system) for 44 to 48 h. After maturation culture, oocytes
were co-incubated with frozen-thawed spermatozoa for 5 h and then cultured for 7 days. The
total mean rates of sperm penetration, normal fertilization, male pronucleus (MPN)
formation, cleavage, and development to the blastocyst stage of oocytes after insemination
were significantly higher (P<0.01) in the rotating culture system than in the static
culture system. In conclusion, compared with the static culture system, the rotating
culture system is adequate for the production of developmentally competent porcine oocytes
when MpFF is used as a maturation medium. 相似文献
20.
Toshinori OIKAWA Tomoko ITAHASHI Takashi NUMABE 《The Journal of reproduction and development》2016,62(1):11-16
The purpose of this study was to determine whether dithiothreitol (DTT) treatment of sperm and ethanol
activation improve embryo production by intracytoplasmic sperm injection (ICSI). Further, we compared ICSI
with standard in vitro fertilization (IVF) in oocytes obtained from cattle. We demonstrated
that DTT reduced the disulfide bond in the bovine sperm head. Using oocytes obtained from a slaughterhouse,
ICSI-DTT treatment without ethanol showed the highest rate of blastocyst formation. We applied these results
to fertilization using ovum pick-up (OPU). Eleven Japanese black cattle served as donors for OPU plus standard
IVF (OPU-IVF). Of them, four donors with low embryo development rates were selected to determine whether
embryo development was enhanced by OPU plus ICSI (OPU-ICSI). We assessed effects on embryo development
following IVF and ICSI in oocytes obtained using OPU. Blastocyst rates were significantly higher for OPU-ICSI
than for OPU-IVF. Our results suggest that OPU-ICSI improves the blastocyst development rate in donors with
low embryo production compared with the standard OPU-IVF. 相似文献