共查询到20条相似文献,搜索用时 609 毫秒
1.
Ji-Yei CHOI Jung-Taek KANG Sol-Ji PARK Su-Jin KIM Joon-Ho MOON Islam M. SAADELDIN Goo JANG Byeong-Chun LEE 《The Journal of reproduction and development》2013,59(5):450-456
One of the factors that impairs in vitro produced porcine embryos
is the oxidative stress that is mainly caused by the imbalance between reactive
oxygen species (ROS) generation and antioxidants activity, especially that of
glutathione (GSH). Here, we examined the effect of 7,8-dihydroxyflavone (7,8-DHF), a
kind of flavonoid antioxidant, on porcine oocyte maturation and its developmental
competence. Porcine oocytes were cultured in media supplemented with 0, 1, 5 and 10
μM 7,8-DHF during both in vitro maturation (IVM) and in
vitro culture (IVC) after parthenogenetic activation. Maturation of
oocytes was evaluated based on first polar body (PB) extrusion and intracellular GSH
level, and developmental competence was assessed through observing cleavage and
blastocyst formation. In each step, the levels of intracellular GSH and ROS were
assessed by fluorescence intensity, and the apoptosis-related gene expression was
examined using semiquantitative RT-PCR. The group treated with 1 μM 7,8-DHF during
IVM and IVC showed increased cytoplasmic maturation and reached the blastocysts stage
(36.1%) at a higher rate than the other groups (24.7, 16.0 and 10.3% for 0, 5 and 10
μM, P<0.05). In that group, the intracellular GSH level was significantly
increased while ROS generation was significantly decreased after IVM and IVC
(P<0.05). Moreover, it showed high expression of an anti-apoptotic gene
(BCL2L1) and low expression of a pro-apoptotic gene
(BAK1) (P<0.05). In conclusion, treatment with 1 μM 7,8-DHF
during IVM and IVC showed an anti-apoptotic effect by increasing intracellular GSH
synthesis and scavenging ROS and therefore improved the developmental competence of
porcine embryos. 相似文献
2.
Weiping HUANG Masashi NAGANO Sung-Sik KANG Yojiro YANAGAWA Yoshiyuki TAKAHASHI 《The Journal of reproduction and development》2014,60(1):9-13
The objective of this study was to clarify the effects of prematurational culture
(pre-IVM) supplemented with 3-isobutyl-1-methylxanthine (IBMX) on nuclear and cytoplasmic
maturation of in vitro-grown bovine oocytes. In experiment 1, oocytes (95
μm in diameter) derived from early antral follicles (0.5–1 mm in diameter) were cultured
for 12 days for in vitro growth (IVG). IVG oocytes with a normal
appearance were subjected to examinations of diameter and chromatin structure in the
germinal vesicle (GV) before IVM. In addition, percentages of metaphase II (M II) were
examined after IVM. Regardless of pre-IVM, the mean diameters of IVG oocytes were about
115 μm. The proportions of GV3 (50.0%) and M II stages (80.1%) of IVG oocytes with pre-IVM
were higher than those without pre-IVM (28.0 and 49.4%, respectively). In experiment 2,
the fertilizability and developmental competence of IVG oocytes were examined. Regardless
of pre-IVM, the normal fertilization rates of IVG oocytes were similar (around 70%) but
were lower than that of in vivo-grown oocytes (88.0%). Cleavage and
blastocyst rates of IVG oocytes with pre-IVM (63.0 and 26.1%, respectively) were higher
than those without pre-IVM (45.8 and 12.7%, respectively). The blastocyst rate based on
cleaved IVG oocytes with pre-IVM (41.7%) was similar to that of in
vivo-grown oocytes (48.7%), although the cleavage rate of IVG oocytes with
pre-IVM was lower than that of in vivo-grown oocytes. In conclusion,
pre-IVM with IBMX improved the maturational and developmental competences of IVG oocytes,
probably due to promotion of their chromatin transition and synchronization of meiotic
progression. 相似文献
3.
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
4.
Misa HOSOE Nao YOSHIDA Yutaka HASHIYADA Hidetoshi TERAMOTO Toru TAKAHASHI Sueo NIIMURA 《The Journal of reproduction and development》2014,60(4):268-273
Fetal bovine serum (FBS) has been widely used as a supplement in the maturation medium of bovine oocytes in
vitro. However, serum contains many undefined factors and is potentially infectious to humans and animals. As a serum
replacement, we evaluated the feasibility of using the silk protein, sericin, derived from the cocoons of silkworm. To examine the
rates of oocyte maturation and fertilization, cumulus-oocyte complexes were cultured in TCM-199 supplemented with 0.01%, 0.05%,
0.1% or 0.15% sericin or 5% FBS. The sizes of the perivitelline space that might relate to polyspermy, the expressions of
Has2 and CD44 mRNA, the amount of hyaluronan (hyaluronic acid: HA) contained in the oocytes
and the rates of blastocyst formation following insemination were then compared between the oocytes cultured with 0.05% sericin
and 5% FBS, because the polyspermy rates in oocytes cultured with 0.05% sericin were significantly lower than in those cultured
with 5% FBS. After in vitro maturation (IVM), the mean size of the perivitelline space was significantly greater
in oocytes cultured with sericin than in those cultured with FBS, although the rates of nuclear maturation, fertilization and
blastocyst formation of oocytes under both IVM conditions were not significantly different. The expression of HAS2 and
CD44 mRNA and the amount of HA in the denuded oocytes cultured with 0.05% sericin were significantly greater than in
those cultured with FBS. These results indicate the feasibility of sericin as an alternative protein supplement for IVM in bovine
oocytes. 相似文献
5.
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
6.
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
7.
Agnieszka Nowak Joanna Kochan Krzysztof Papis Adam Okólski 《Journal of Equine Veterinary Science》2014
The aim of the present study was to determine the vitality and developmental competence of equine oocytes after in vitro maturation (IVM) and vitrified by Rapid-i method. In experiment 1, oocytes after IVM were vitrified using media: EquiPro VitKit (group 1) or medium containing 18% Ficoll, 40% ethylene glycol, and 0.3 M sucrose (group 2). For evaluation of toxicity effect, oocytes were exposed to media without a plug to liquid nitrogen. To evaluate viability, oocytes were stained with fluorescein diacetate and ethidium bromide. In experiment 2, oocytes after IVM and vitrification were activated by 7.5 μM ionomicin in TCM 199 (5 minutes) combined with 2 mM 6-DMAP in TCM 199 with 10% fetal bovine serum (4.5 hours). Survival rate was: 63% in group 1 (n = 54), 55% in group 2 (n = 69), and 73.2% (n = 56) in the control group. After parthenogenetic activation, 10.2% (n = 49) of 2–4 blastomeres were observed. This percentage was lower than in the nonvitrified group: 38.5% (n = 53). 相似文献
8.
Vibuntita CHANKITISAKUL Nutthee AM-IN Theerawat THARASANIT Tamas SOMFAI Takashi NAGAI Mongkol TECHAKUMPHU 《The Journal of reproduction and development》2013,59(1):66-71
Failure of male pronucleus formation has hampered the success of intracytoplasmic sperm
injection (ICSI) in swamp buffalo. The aim of the present study was to improve male
pronucleus formation by pretreating sperm with various chemicals before ICSI. In
Experiments1 and 2, sperm were treated according to one of the following protocols: (1)
0.1% Triton-X 100 (TX) for 1 min, (2) 10 µM calcium ionophore (CaI) for 20 min, (3)
freezing and thawing (FT) without any cryoprotectant, or (4) no treatment (control). These
sperm treatment groups then either did or did not receive additional sperm treatment with
5 mM dithiothreitol (DTT) for 20 min. Acrosomal integrity (Experiment 1) and DNA
fragmentation (Experiment 2) were evaluated in the sperm before ICSI. In Experiment 3,
oocytes matured in vitro were subjected to ICSI using pretreated sperm as
described above and then were cultured either with or without activation. The TX- and
CaI-treated sperm caused an increase in the number of acrosome-loss sperm, whereas the FT
treatment and control increased the proportion of acrosome-reacted sperm (P<0.05). The
DNA fragmentation did not differ among treatments (P>0.05). At 18 h post-ICSI,
pronucleus (PN) formation was found only in activated oocytes. The majority of the
activated ICSI oocytes contained intact sperm heads. Normal fertilization was observed in
the CaI and FT treatment groups and control group when sperm were treated with DTT before
ICSI. In conclusion, DTT treatment of sperm with reacted acrosomes before ICSI together
with activation of the ICSI oocytes is important for successful male pronucleus
formation. 相似文献
9.
10.
Sandra Soto‐Heras Maria‐Gracia Catal Montserrat Roura Irene Menndez‐Blanco Anna‐Rita Piras Dolors Izquierdo Maria‐Teresa Paramio 《Reproduction in domestic animals》2019,54(2):381-390
Melatonin enhances in vitro embryo development in several species by improving the oocyte developmental competence during in vitro maturation (IVM). Melatonin has a wide range of actions, from scavenging reactive oxygen species (ROS) to regulating gene expression, and it can also act by way of melatonin receptors. The aim of this study was to determine the mechanism of action of melatonin during the IVM of juvenile goat oocytes and the role of the membrane receptors. Melatonin receptor 1 was immunolocalized in cumulus cells and oocytes before and after 24 hr of IVM. The effect of melatonin on oocyte developmental competence was tested in three experimental IVM groups: (a) control, (b) 10?7 M melatonin, and (c) 10?7 M melatonin +10?7 M luzindole (an inhibitor of both melatonin receptors). After IVM oocytes were assessed for ROS levels, mitochondrial activity, adenosine 5′‐triphosphate (ATP) concentration and relative gene expression (ACTB, SLC1A1, SOD1, GPx1, BAX, DNMT1, GCLC and GDF9). IVM‐oocytes were in vitro fertilized and cultured under conventional conditions. Blastocyst rate and quality (differential cell count) were assessed at 8 days post‐fertilization. Melatonin decreased ROS levels, increased mitochondrial activity and ATP content and increased blastocyst quality compared to control group (55.8 vs. 30.4 inner cell mass ICM, p < 0.05). There was no effect on the relative gene expression due to treatment with melatonin. In conclusion, we have showed that melatonin improves oocyte developmental competence in juvenile goats by reducing ROS levels and improving mitochondrial activity. 相似文献
11.
《中国畜牧杂志》2019,(10)
本研究旨在探讨锌对牛卵母细胞体外成熟及体外受精胚胎发育的影响。首先使用锌螯合剂TPEN去除锌,并检测缺锌对牛卵母细胞体外成熟的影响;然后在体外成熟液中分别添加0(对照组)、0.4、0.8、1.2、1.6μg/mL硫酸锌,探索不同浓度硫酸锌对体外成熟及后续胚胎发育的影响。结果表明:锌元素在体外成熟液中的含量低于牛卵泡液和颈静脉血清(P<0.05);去除体外成熟液中的锌后牛卵母细胞的体外成熟效率下降(P<0.05),且具有时间依赖性,补充适宜浓度硫酸锌后成熟效率恢复;向体外成熟液中添加硫酸锌并未对卵母细胞体外成熟效率产生显著影响,但添加0.8μg/mL硫酸锌成熟后的卵母细胞中活性氧含量显著降低,后续体外受精胚胎的囊胚发育效率显著提高;RT-qPCR分析结果显示,与对照组相比,添加0.8μg/mL硫酸锌成熟后的卵母细胞中抗氧化基因SOD1、CAT、TXN1、PRD1和卵丘扩展基因PTX3、TSG6的表达水平均提高(P<0.05)。研究表明,添加0.8μg/mL硫酸锌可以通过提高卵母细胞内抗氧化酶基因的表达水平,降低卵母细胞内活性氧含量,促进卵丘扩展,从而提高卵母细胞成熟质量和体外受精胚胎的发育效率。 相似文献
12.
Experimental approach to prezygotic chromosome screening using only a single pair of gametes in
mice
Hiroyuki WATANABE Atsushi KOHDA Hiroyuki TATENO 《The Journal of reproduction and development》2015,61(6):511-518
During in vitro embryo production, chromosome screening is essential to prevent pregnancy
losses caused by embryonic chromosome aberrations. When the chromosome screening is completed before
fertilization, gametes are effectively utilized as genetic resources. The aim of this study was to investigate
whether chromosome screening of gametes accompanied by fertilization would be feasible using a single mouse
spermatozoon and oocyte. Metaphase II oocytes were divided into a cytoplast and a karyoplast. For genome
cloning of the gametes, androgenic and gynogenic embryos were produced by microinjection of sperm into
cytoplasts and parthenogenetic activation of karyoplasts, respectively. Pairs of blastomeres from androgenic
and gynogenic embryos were fused electrically to produce diploid embryos, which were transferred into
pseudopregnant surrogate mothers to examine fetal development. Blastomeres from androgenic and gynogenic
embryos were individually treated with calyculin A—a specific inhibitor of type 1 and 2A protein
phosphatases—for 2 h to induce premature chromosome condensation. Thereafter, chromosome analysis of
blastomeres, reflecting the genetic constitution of individual spermatozoa and oocytes, was performed, and we
confirmed that most of the androgenic and gynogenic 2-cell embryos had a haploid set of chromosomes in their
sister blastomeres. The reconstructed embryos from blastomeres of androgenic and gynogenic 2-cell embryos
could be implanted and develop into live fetuses, albeit at low efficiency. This study indicates that
prezygotic chromosome screening and embryo production using a single pair of gametes may be practicable. 相似文献
13.
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
14.
试验旨在探讨藏红花素(crocin)的抗氧化应激作用对小鼠卵母细胞体外成熟及后续胚胎发育能力的影响。在体外成熟(IVM)培养液中添加不同浓度藏红花素(0、5、10、15、20、25、30 μmol/L),小鼠卵母细胞在体外成熟培养12 h后,检测卵母细胞第一极排出情况、卵母细胞胞质内活性氧(ROS)和谷胱甘肽(GSH)含量,并进行体外受精(IVF);体外受精后6 h统计受精率,24 h统计卵裂率,96 h统计囊胚率。结果显示,与对照组相比,10、15 μmol/L藏红花素显著提高了卵母细胞第一极体排出率(P<0.05);当藏红花素浓度继续增加时,卵母细胞的第一极体排出率下降,30 μmol/L藏红花素显著降低了卵母细胞第一极体排出率(P<0.05);5、10、15 μmol/L藏红花素均显著降低了卵母细胞ROS含量(P<0.05),10、15 μmol/L藏红花素均显著提高了卵母细胞GSH含量(P<0.05)。与对照组相比,10 μmol/L藏红花素组受精率、卵裂率、囊胚率差异均不显著(P>0.05),15、20、25、30 μmol/L藏红花素均显著降低受精率和囊胚率(P<0.05),对卵裂率影响不显著(P>0.05)。结果表明,在小鼠卵母细胞体外成熟培养液中添加10 μmol/L藏红花素可以显著增加第一极体排出率,显著降低卵母细胞ROS含量、提高卵母细胞GSH含量,但对受精后的胚胎发育无显著影响。 相似文献
15.
为了初步了解组织型纤溶酶原激活剂(tissue-plasminogen activator,tPA)在牛卵丘-卵母复合体体外成熟进程中的潜在作用,试验运用实时定量RT-PCR技术分别检测体外成熟过程中不同时间段(0、8、16、24 h)以及添加不同浓度FSH成熟培养16 h后的牛卵丘细胞和卵母细胞中tPA基因相对表达变化,观察添加不同浓度FSH成熟培养16 h后卵丘细胞膨胀程度差异,并统计卵母细胞第一极体排出情况。结果显示,卵丘-卵母复合体在体外成熟初期,tPA mRNA相对表达水平在体外成熟16和24 h的卵丘细胞和卵母细胞中显著高于0和8 h组;在添加不同浓度FSH (0、0.01、0.1和1 IU/mL)体外成熟16 h的各处理组,卵丘细胞中tPA mRNA相对表达量随FSH添加浓度的升高而升高,卵丘细胞的扩散程度亦随之增加;同时tPA mRNA相对表达量在0.01 IU/mL FSH添加组卵母细胞中显著高于其他各FSH添加组。综合以上研究结果,本研究认为,牛卵丘细胞中tPA基因的表达受FSH正向调节;tPA可能促进了卵丘细胞在体外成熟过程中的扩散;同时,tPA在卵母细胞中的表达是否与其第一极体排出有关,需进一步试验验证。 相似文献
16.
Kanchana PUNYAWAI Nitira ANAKKUL Kanokwan SRIRATTANA Yoshio AIKAWA Siwat SANGSRITAVONG Takashi NAGAI Kei IMAI Rangsun PARNPAI 《The Journal of reproduction and development》2015,61(5):431-437
This study was designed to compare the efficiency of the Cryotop method and that of two methods that employ a micro volume air cooling (MVAC) device by analyzing the survival and development of bovine oocytes and blastocysts vitrified using each method. In experiment I, in vitro-matured (IVM) oocytes were vitrified using an MVAC device without direct contact with liquid nitrogen (LN2; MVAC group) or directly plunged into LN2 (MVAC in LN2 group). A third group of IVM oocytes was vitrified using a Cryotop device (Cryotop group). After warming, vitrified oocytes were fertilized in vitro. There were no significant differences in cleavage and blastocyst formation rates among the three vitrified groups, with the rates ranging from 53.1% to 56.6% and 20.0% to 25.5%, respectively; however, the rates were significantly lower (P < 0.05) than those of the fresh control group (89.3% and 43.3%, respectively) and the
solution control group (87.3% and 42.0%, respectively). In experiment II, in vitro-produced (IVP) expanded blastocysts were vitrified using the MVAC, MVAC in LN2 and Cryotop methods, warmed and cultured for survival analysis and then compared with the solution control group. The rate of development of vitrified-warmed expanded blastocysts to the hatched blastocyst stage after 24 h of culture was lower in the MVAC in LN2 group than in the solution control group; however, after 48–72 h of culture, the rates did not significantly differ between the groups. These results indicate that the MVAC method without direct LN2 contact is as effective as the standard Cryotop method for vitrification of bovine IVM oocytes and IVP expanded blastocysts. 相似文献
17.
Teewara Phongnimitr Yuanyuan Liang Kanokwan Srirattana Kanchana Panyawai Nucharin Sripunya Chatchai Treetampinich Rangsun Parnpai 《Animal Science Journal》2013,84(11):719-725
In this study, the effects of the addition of L‐carnitine in in vitro maturation (IVM) medium for bovine oocytes on their nuclear maturation and cryopreservation were investigated; they were matured in IVM medium supplemented with 0.0, 0.3, 0.6 and 1.2 mg/mL of L‐carnitine (control, 0.3, 0.6 and 1.2 groups, respectively) and some of them were vitrified by Cryotop. Moreover, the effects of L‐carnitine during in vitro fertilization (IVF) and in vitro culture (IVC) on the developmental potential and quality of IVF embryos were also examined. A significantly higher maturation rate of oocytes was obtained for 0.3 and 0.6 mg/mL groups compared with the control (P < 0.05). The blastocyst formation rate in the 0.6 group was significantly improved, whereas the rate in the 1.2 group was significantly decreased when compared with the control group (P < 0.05). No significant difference was found in embryo development between the control and the L‐carnitine group after oocyte vitrification. Supplementation of IVF and IVC media with L‐carnitine had no effect on development to the blastocyst stage of IVM oocytes treated with 0.6 mg/mL L‐carnitine. In conclusion, the supplementation of L‐carnitine during IVM of bovine oocytes improved their nuclear maturation and subsequent embryo development after IVF, but when they were vitrified the improving effects were neutralized. 相似文献
18.
【目的】研究柠檬苦素(limonin,Lim)对小鼠卵母细胞体外成熟(IVM)及后续体外受精(IVF)胚胎发育潜能的影响,旨在为体外成熟培养系统的优化提供参考。【方法】在小鼠体外成熟培养液中添加不同浓度的Lim(0、10、20、50 μmol/L),成熟培养12 h后统计小鼠卵母细胞第一极体(PBI)排出率,筛选体外成熟培养液中添加Lim的最适浓度;在体外成熟培养液中添加最适浓度的Lim,以0 μmol/L Lim为对照组,成熟培养12 h,通过免疫荧光染色检测活性氧(ROS)、谷胱甘肽(GSH)以及线粒体膜电位(MMP)水平;通过实时荧光定量PCR检测卵母细胞抗氧化及凋亡相关基因的mRNA表达水平。将最适Lim组及对照组卵母细胞体外成熟24 h后进行体外受精,于体外受精24 h和3.5 d分别统计胚胎卵裂率和囊胚率,并用Fluorescein-dUTP和Hoechst 33342染色分别检测囊胚总细胞数及囊胚内凋亡细胞比率。【结果】与0 μmol/L Lim组相比,20 μmol/L Lim组小鼠卵母细胞PBI排出率显著升高(P<0.05),后续试验均用20 μmol/L Lim进行处理。与对照组组相比,20 μmol/L Lim组小鼠卵母细胞内ROS水平显著降低(P<0.05),GSH、MMP水平均显著增加(P<0.05),抗氧化相关基因(GPx3、CAT和Prdx3)、抗凋亡相关基因(Bcl-2、Bcl-xl)表达水平均显著上调(P<0.05),促凋亡相关基因(Caspase-3)表达水平显著下调(P<0.05);体外受精胚胎的卵裂率、囊胚率、囊胚总细胞数均显著增加(P<0.05),囊胚内细胞凋亡比率显著下降(P<0.05)。【结论】在体外成熟培养液中添加20 μmol/L Lim可以通过抑制氧化应激和细胞凋亡、增加MMP水平提高小鼠卵母细胞质量,从而提高体外受精胚胎的发育潜力。 相似文献
19.
Michel KERE Chawalit SIRIBOON Neng-Wen LO Ngoc Tan NGUYEN Jyh-Cherng JU 《The Journal of reproduction and development》2013,59(1):78-84
In this study, a dose-response assessment was performed to understand the relation
between supplementation of media with L-ascorbic acid or vitamin C and porcine oocyte
maturation and the in vitro development of parthenotes (PA) and handmade
cloned (HMC) embryos. Various concentrations (0, 25, 50 and 100 µg/ml) of vitamin C
supplemented in in vitro maturation (IVM) and culture (IVC) media were
tested. None of these vitamin C additions affected nuclear maturation of oocytes, yet
supplementation at 50 µg/ml led to significantly increased intracellular glutathione (GSH)
levels and reduced reactive oxygen species (ROS). When cultured in IVM- and/or
IVC-supplemented media, the group supplemented with 50 µg/ml of vitamin C showed improved
cleavage rates, blastocyst rates and total cell numbers per blastocyst (P<0.05)
compared with other groups (control, 25 µg/ml and 100 µg/ml). In contrast, supplementation
with 50 µg/ml vitamin C decreased (P<0.05) the apoptosis index as compared with the
groups supplemented with 100 µg/ml. In addition, even with a lower blastocyst rate to
start with (37.6 vs. 50.3%, P<0.05), supplementation of HMC embryos
with vitamin C ameliorated their blastocyst quality to the extent of PA embryos as
indicated by their total cell numbers (61.2 vs. 59.1). Taken together, an
optimized concentration of vitamin C supplementation in the medium not only improves
blastocyst rates and total cell numbers but also reduces apoptotic indices, whereas
overdosages compromise various aspects of the development of parthenotes and cloned
porcine embryos. 相似文献
20.
Transporting bovine oocytes in a medium supplemented with different macromolecules and antioxidants: Effects on nuclear and cytoplasmic maturation and embryonic development in vitro 下载免费PDF全文
M Ambrogi PC Dall'Acqua NAS Rocha‐Frigoni BCS Leão GZ Mingoti 《Reproduction in domestic animals》2017,52(3):409-421
We investigated whether supplementing the medium used to transport bovine oocytes with different macromolecules [foetal calf serum (FCS) or bovine serum albumin (BSA)] or a mixture of antioxidants (cysteine, cysteamine and catalase) affects their nuclear and cytoplasmic maturation and thereby affects their subsequent embryonic development and cryotolerance. Oocytes were transported for 6 hr in a portable incubator and then subjected to standard in vitro maturation (IVM) for 18 hr. The oocytes in the control groups were cultured (standard IVM) for 24 hr in medium containing 10% FCS (Control FCS) or 10% FCS and the antioxidant mixture (Control FCS+Antiox). The intracellular concentrations of reactive oxygen species (ROS) at the end of IVM period were lower in the oocytes subjected to simulated transport in the presence of a macromolecular supplement or the antioxidant mixture than that of the control group (FCS: 0.62 and BSA: 0.66 vs. Control FCS: 1.00, p < .05; and Transp: 0.58 and Transp Antiox: 0.70 vs. Control FCS: 1.00, p < .05). After IVM, the mitochondrial membrane potentials of the transported oocytes were lower than those of the non‐transported oocytes (FCS: 0.41 and BSA: 0.57 vs. Control FCS: 1.00, p < .05; and Transp: 0.48 and Transp Antiox: 0.51 vs. Control FCS: 1.00 and Control Antiox: 0.84, p < .05). The blastocyst formation rates (36.9% average) and the re‐expansion rates of vitrified‐warmed blastocysts (53%, average) were unaffected (p > .05) by the treatments. In conclusion, supplementing the medium in which bovine oocytes are transported with antioxidants or different macromolecules did not affect their in vitro production of embryos or their cryotolerance. 相似文献