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1.
In vitro Development of Buffalo Oocytes in Media-containing Fluids from Different Size Class Follicles 总被引：1，自引：0，他引：1
S Nandi HM Raghu BM Ravindranatha PSP Gupta PV Sarma 《Reproduction in domestic animals》2004,39(1):33-38
Studies were conducted to investigate the effect of supplementation of fluid from different sized class [small (SFF, < 3 mm), medium (MFF, 3-8 mm) and large (LFF, > 8 mm)] of normal and cystic (CFF) ovarian follicles in oocyte culture media on oocyte maturation rate and embryo development in vitro and to test the efficacy of follicular fluid (FF) from different size classes as a whole oocyte maturation medium. Results suggested that FF were capable of developing buffalo oocytes to embryonic stage in vitro although its efficacy was lower than that of serum. Regardless of high maturation rates after in vitro maturation (IVM) in media containing FF or IVM in whole FF, low blastocyst rates were obtained after in vitro fertilization (IVF) and culture of embryos. Follicular fluid from small follicles had significantly (p < 0.05) higher potential of developing buffalo oocytes to embryonic stage in vitro than that from medium and large follicles. Cystic FF was not capable of supporting development of buffalo oocytes in vitro. 相似文献
2.
A. Krogences E. Ropstad T Nilsen
. Pedersen M. Forsberg 《Acta veterinaria Scandinavica》1993,34(2):211
In vitro maturation of oocytes has been described for several species, but to our knowledge this is the first attempt to mature oocytes from reindeer in vitro. Reindeer are seasonal breeders with their main period of breeding activity starting in the beginning of October. Little knowledge exists about in vivo oocyte maturation, fertilization and development of the early embryonic stages after fertilization. The aim of the present study was to examine whether reindeer oocytes collected out of breeding season could be matured in vitro in conventional medias used for bovine oocytes. 相似文献
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This communication describes the in vitro maturation and in vitro fertilization of bovine follicular oocytes. Further development of the embryos was achieved by using a granulosa cell culture system. The in vitro development of oocytes to morula/blastocyst stages obtained from individual cows was compared to the results of pooled simultaneous cultured oocytes and to our over-all results of this method. While there were no statistical differences in the developmental rates between these three groups (individual cows: 28.1%, simultaneous pool: 34.0%, over-all results: 32.7%) marked differences were found between the 22 animals investigated separately. These results indicate that there were great individual variabilities due to the oocyte population comparable to the variations in ovarian response to superovulation. 相似文献
5.
本研究观察了不同超排处理方法对羔山羊卵母细胞体外成熟及体外受精的影响，并将体外受精胚胎进行了移植，以便探讨利用羔山羊卵母细胞生产体外受精胚胎的技术途径。对２～４月龄羔山羊用ＦＳＨ、ＦＳＨ＋ＬＨ（静脉注射）和ＦＳＨ＋ＬＨ（肌肉注射）三种方法进行超排处理，平均分别回收１６．０、２０．３和１５．０枚卵母细胞。将这些卵母细胞进行２０～２４ｈ的成熟培养后，其成熟率分别为４５．５％，４４．４％和７６．５％。成熟卵母细胞经体外受精，体外发育培养后其２～４细胞胚胎的发生率分别为５．６％ａ、６．７％和３１．７％ｂ（ｂ，ｖｓ，ａ，Ｐ＜０．０５）。将９枚２～４细胞期胚胎移植给６只受体母羊，结果有２只妊娠并产羔２只。 相似文献
6.
《Domestic animal endocrinology》1994,11(1):59-86
Crossbred heifers (n = 103) were synchronized to estrus with prostaglandin (PGF2α) and superovulated with follicle stimulating hormone (FSH-P). Animals were ovariectomized every 12 hr after the PGF2α injection (n = 7 to 9/time) up to 108 hr to monitor the follicular, hormonal, and oocyte changes associated with follicular development and ovulation. Twenty-eight animals were implanted with Norgestomet implants 12 hr before PGF2α and ovariectomized at 72, 84, 96, and 108 hr post PGF2α injection to monitor effects of progesterone and suppression of the luteinizing hormone (LH) surge on oocyte maturation and quality. Follicular fluid was collected and analyzed for progesterone, estradiol, prolactin, and glycosaminoglycan content in conjunction with cumulus maturation and nuclear stage of oocyte maturation. Analysis of in vivo matured oocytes by in vitro fertilization was carried out at 60, 72, 84, and 96 hr post PGF2α and in vitro matured oocytes at 12 to 108 hr post PGF2α. No developmental changes in cumulus cells surrounding the oocyte of small follicles was noted (≤ 4 mm dia) indicating a static population. Medium (> 4 ≤ 8 mm) and large size (> 8 mm) follicles developed to the corona radiata and loose cumulus stages in animals in which an LH surge was detected but cumulus status remained primarily in the tight cumulus stage for animals without an LH surge. The estradiol-to-progesterone ratio for tight cumulus (TC), corona radiata (CR), and loose cumulus (LC) stages was 1.8 ± .1, 1.0 ± .1, and .4 ± .2, respectively (P < .01). Nuclear maturation of oocytes in small follicles from animals without a detectable LH surge seem to indicate early maturation (48 to 72 hr post PGF2α) in conjunction with a high percent of degenerate oocytes not seen in animals exhibiting an LH surge. Oocytes from medium size follicles matured to germinal vesicle breakdown (GVBD) and early meiosis (metaphase I; MI) stages of development in all treatments. Most oocytes were degenerate in Norgestomet-implanted animals. Oocytes from large follicles (> 8 mm dia) from animals exhibiting an LH surge were in MI and metaphase II (MII) stages (48 to 84 hr post PGF2α) in preparation of ovulation whereas oocytes from animals not exhibiting an LH surge had oocytes that early matured to MII (48 to 72 hr post PGF2α), later regressing to degenerate oocytes (84 to 108 hr). Follicular progesterone, estradiol, and prolactin increased with oocyte maturation, particularly in medium and large follicles. In vivo matured oocytes for fertilization (60, 72, 84, and 96 hr post PGF2α) were nude (from the oviduct) and primarily CR from follicles. Tubal oocytes (37%) were fertilized more frequently by a single sperm than follicular oocytes (14.3%; P < .01) and single sperm penetration peaked at 72 hr post PGF2α. Follicular hormone concentrations were not related to sperm penetration. Oocytes (n = 101) matured in vivo had lower fertilization potential from ovaries producing < 14 or > 50 follicles (39.3%) as compared to 21 to 45 aspirated follicles (68.2%; P < .05), with a peak penetration at 32 follicles (86.7% penetration). No treatment differences (LH surge or no detectable LH surge) were noted in relation to in vivo matured oocytes. Oocytes with single sperm penetration had the lowest estradiol/progesterone ratio of 2.2 vs polyspermic penetration of 13.7. 相似文献
7.
A study was conducted opportunistically to evaluate the potential of rescuing immature oocytes from the ovaries of the Sumatran rhinoceros postmortem. Recovered oocytes (n = 30) were placed in maturation culture for 36 hr and inseminated with frozen-thawed homologous spermatozoa. After culture, evaluation of nuclear maturation status revealed that a large number of oocytes were degenerated (n = 21), but nine oocytes were assessed at the germinal vesicle (n = 3), metaphase I (n = 3), and metaphase II (n = 3) stages. Frozen-thawed Sumatran rhinoceros spermatozoa were capable of binding to the zona pellucida of in vitro matured oocytes, but no fertilization or cleavage resulted. In conclusion, relatively large numbers of oocytes can be obtained by ovarian follicular aspiration postmortem in the Sumatran rhinoceros, and some of these oocytes are capable of achieving nuclear maturation in vitro. However, additional studies are required to improve maturation success and achieve fertilization in culture. 相似文献
8.
为了解水牛卵母细胞和体外受精（IVF）胚胎早期发育过程中端粒酶的活性变化,本研究利用端粒重复序列扩增法（TRAP）进行了水牛未成熟卵母细胞,成熟卵母细胞和2～4细胞,8～16细胞,桑椹胚以及囊胚各阶段的早期胚胎端粒酶活性的测定。依据电泳条带在成像系统下的光密度值,计算端粒酶的相对活性（RTA）。结果发现,未成熟卵母细胞端粒酶活性比成熟卵母细胞高（P〈0.05）,受精后2～4和8～16细胞胚胎端粒酶活性相对较低,桑椹胚端粒酶活性明显升高（P〈0.05）,囊胚阶段达到最高水平。通过对水牛不同发育阶段胚胎细胞数计数及单细胞相对端粒酶活性的分析比较结果显示,卵母细胞的单细胞端粒酶活性最高,囊胚阶段的最低。单细胞端粒酶活性从未成熟卵母细胞到IVF囊胚阶段呈逐渐降低的趋势。这些结果表明,水牛卵母细胞及早期胚胎的端粒酶活性变化与其成熟、发育阻断及全能性的逐步降低有关。 相似文献
9.
Hui PENG Xiujiao LIN Fang LIU Cheng WANG Wenchang ZHANG 《The Journal of reproduction and development》2015,61(6):559-564
Nlrp9a, Nlrp9b and Nlrp9c are preferentially expressed in
oocytes and early embryos in the mouse. Simultaneous genetic ablation of Nlrp9a and
Nlrp9c does not affect early embryonic development, but the function of
Nlrp9b in the process of oocyte maturation and embryonic development has not been
elucidated. Here we show that both Nlrp9b mRNA and its protein are expressed in ovaries and
the small intestine. Moreover, the NLRP9B protein was restricted to oocytes in the ovary and declined with
oocyte aging. After ovulation and fertilization, NLRP9B protein was found in preimplantation embryos. Confocal
microscopy demonstrated that it was mainly localized in the cytoplasm in the oocytes and blastomeres. Thus,
this protein might play a role in oocyte maturation and early embryonic development. However, knockdown of
Nlrp9b expression in GV-stage oocytes using RNA interference did not affect oocyte
maturation or subsequent parthenogenetic development after Nlrp9b-deficient oocytes were
activated. Furthermore, Nlrp9b knockdown zygotes could reach the blastocyst stage after being
cultured for 3.5 days in vitro. These results provide the first evidence that the NLRP9B
protein is dispensable for oocyte maturation and early embryonic development in the mouse. 相似文献
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The effect of source of cumulus-oocytes-complexes (COCs), maturation and fertilization conditions on developmental competence of dromedary embryos was examined. Thirty-six adult females were superovulated with equine Chorionic Gonadotropin (eCG) injection (3500 IU, IM) and divided in three groups of 12 females each. Group 1 provided 138 COC's collected from follicles >or= 5 mm 10 days after stimulation prior hCG treatment and matured in vitro for 30 h. Group 2 provided 120 in vivo matured oocytes which were aspirated from their follicles 20 h after hCG (3000 IU, IV) given on day 10 follow eCG injection. Group 3 provided 65 in vivo matured/fertilized oocytes. Females in Group 3 received hCG on day 10 following eCG treatment and then were mated 24 h later. Fertilized oocytes were collected from the oviducts of females 48-h post-mating. Quality of the oocytes was assessed after in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC) of COCs. All cultures were performed in three replicates (n = 3) at 38.5 degrees C, under 5% CO(2) and high humidity (>95%). Only COCs with cumulus and homogenous (dark) cytoplasm were used. Nuclear maturation rate for Groups 1 and 2 was determined by epifluorescence microscopy in a sample of COCs (n = 30) denuded, fixed and stained with Hoechst 33342. To study the viability of obtained embryos, hatched blastocysts from each group were transferred to recipients followed by pregnancy diagnosis using ultrasonography at 15, 60 and 90 days. The percentage of COCs reaching metaphase II (MII) after 30 h of maturation was slightly but not significantly higher for in vivo matured oocytes (28/30; 93%) than those in vitro matured (25/30; 84%). The total rate of cleavage (2 cells to blastocyst stage) was not different for the three groups. However, significantly (p < 0.05) more blastocyst and hatched blastocysts were obtained from in vivo matured and in vivo fertilized oocytes (Group 3; 52% and 73%) than from in vitro fertilized oocytes whether they were matured in vitro (Group 1; 35% and 32%) or in vivo (Group 2; 32% and 45%). Pregnancy rates were not significantly different amongst all groups for the three first months following embryo transfer. All pregnancies were lost after day 90 follow transfer except for in vivo matured and in vivo matured/fertilized groups. Only in vivo matured/in vitro fertilized and in vivo matured/fertilized produced embryos continued normal development until term and resulted in the birth of normal and healthy live calves. Six claves (29%; 6/21) were born from Group 3 and one (8%; 1/13) calf was born from Group 2. This study shows that the IVC system used is able to support camel embryo development. However, developmental competence and viability of dromedary embryos may be directly related to the intrinsic quality (cytoplasmic maturation) of oocytes. 相似文献
12.
Chlorogenic acid supplementation during in vitro maturation improves maturation,fertilization and developmental competence of porcine oocytes 
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下载免费PDF全文 T‐V Nguyen F Tanihara LTK Do Y Sato M Taniguchi M Takagi T Van Nguyen T Otoi 《Reproduction in domestic animals》2017,52(6):969-975
Chlorogenic acid (CGA) is a quinic acid conjugate of caffeic acid, and a phytochemical found in many fruits and beverages that acts as an antioxidant. The present study investigated the effects of CGA supplementation during in vitro maturation (IVM), on in vitro development of porcine oocytes, to improve the porcine in vitro production (IVP) system. Oocytes were matured either without (control) or with CGA (10, 50, 100 and 200 μM). Subsequently, the matured oocytes were fertilized and cultured in vitro for 7 day. The rates of maturation, fertilization and blastocyst formation of oocytes matured with 50 μM CGA were significantly (p < .05) higher than those of the control oocytes. Hydrogen peroxide (H2O2) is one of the reactive oxygen species and induces DNA damage in porcine oocytes. When oocytes were matured with 1 mM H2O2 to assess the protective effect of CGA, 50 μM CGA supplementation improved the maturation rate and the proportion of DNA‐fragmented nuclei in oocytes compared with control oocytes matured without CGA. Moreover, when oocytes were matured with either 50 μM CGA (control) or caffeic acid (10, 50 and 100 μM), the rates of maturation, fertilization and the blastocyst formation of oocytes matured with 50 μM CGA were similar to those of oocytes matured with 10 and 50 μM caffeic acid. Our results suggest that CGA has comparable effects to caffeic acid, and IVM with 50 μM CGA is particularly beneficial to IVP of porcine embryos and protects oocytes from DNA damage induced by oxidative stress. Supplementation of CGA to the maturation medium has a potential to improve porcine IVP system. 相似文献
13.
Fertilization and blastocyst development in oocytes obtained from prepubertal and adult pigs 总被引：2，自引：0，他引：2
Polyspermic fertilization and embryo quality are important issues for the in vitro production of pig embryos. We hypothesized that oocyte donor (prepubertal gilt vs. sow) affects polyspermy and blastocyst development in vitro and that the sexual maturity of the oocyte donor affects the response to sperm concentration in the fertilization medium. In Exp. 1, oocytes of sows and gilts were mounted and stained 12 h after insemination to provide fertilization data. In Exp. 2, putative embryos were developed in vitro to 144 h post-insemination before mounting. In both experiments, cumulus-oocyte complexes (COC) were collected from ovaries of prepubertal gilts and adult sows. Sperm were added after maturation of COC for 40 to 44 h. Sperm from two boars at 0.5 to 5.0 x 10(6) sperm/mL was used for insemination. More (P < 0.01) monospermic fertilizations were observed in oocytes derived from gilts than for oocytes from sows. There were fewer (P < 0.02) penetrated sperm per fertilized oocyte in oocytes from gilts compared with sows. There were effects of semen donor (boar) on the percentage of monospermic (P < 0.01) and polyspermic (P < 0.002) fertilizations, and on the number of penetrated sperm/fertilized oocyte (P < 0.02). In Exp. 2, cleavage and blastocyst formation was evaluated at 2 and 6 d postinsemination, respectively. More (P < 0.001) blastocysts developed from sow-derived oocytes than from gilt-derived oocytes. More (P < 0.05) total cells per blastocyst were observed in embryos from sow-derived oocytes than from gilt-derived oocytes. Semen donor affected the percentage of oocytes cleaving (P < 0.02), and a boar x sperm concentration interaction affected (P < 0.05) the incidence of blastocyt formation. Results indicate that sexual maturity of the donor is not responsible for the high incidence of polyspermy in porcine in vitro fertilization. However, blastocyst development is improved by the use of oocytes from sows rather than from prepubertal gilts. 相似文献
14.
Agung B Otoi T Wongsrikeao P Taniguchi M Shimizu R Watari H Nagai T 《The Journal of reproduction and development》2006,52(1):123-127
It has been suggested that the maturational stage of oocytes at time of insemination influences the sex ratio of resulting embryos. However, there are very few reports concerning the relationship between the maturation culture period of oocytes and the sex ratio of resulting embryos. The objective of this study was to investigate the effects of in vitro maturation culture period for bovine oocytes on the sex ratio of in vitro produced blastocysts using a novel technique of loop-mediated isothermal amplification (LAMP). Cumulus-oocyte complexes were collected from the ovaries of slaughtered cows, and then matured in vitro for various periods (16, 22, 28, and 34 h). After maturation culture for each period, the oocytes were inseminated with frozen-thawed spermatozoa, and then cultured in vitro. Blastocysts were harvested on Day 7 after insemination, and the sex of the embryos was examined using the LAMP method. The rates of oocytes matured to the metaphase II stage were significantly lower (P < 0.05) in the 16-h maturation group than in the other groups. The proportion of blastocyst formation after insemination was significantly higher (P < 0.05) in the 22-h maturation group than in the other groups. The proportion of male blastocysts increased with the increase in maturation culture period. The proportion of male blastocysts derived from oocytes matured for 34 h was significantly higher (P < 0.05) than from oocytes matured for 16 and 22 h. These results indicate that the sex ratio of in vitro fertilized embryos is apparently influenced by the maturation culture period of the oocytes. 相似文献
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SHI Xian XIONG Xian-rong LAN Dao-liang HU Jia-jia CAI Wen-yi ZHANG Da-wei LI Jian 《中国畜牧兽医》2017,44(1):155-160
This study was designed to investigate the effect of different types and different concentrations of sugar on in vitro maturation(IVM) and developmental competence of yak oocytes, for being further research and optimization culture system of yak oocytes for efficient maturity yak oocytes and productivity of embryos. Immature yak oocytes were matured in vitro on culture medium with different concentrations (0,5 and 10 mmol/L) of glucose and sucrose in incubator for 24 h or 2 h pretreament with sugar and 22 h without sugar. Subsequently, then the maturation of oocytes,the cleavage rates and blastocyst formation rates after in vitro fertilization(IVF) were evaluated. The results showed that a medium with 5 and 10 mmol/L glucose IVM could significantly increase the yak oocytes maturation and cleavage (P<0.05), and the highest blastocyst formation rates in 10 mmol/L glucose group was significantly higher than 0 mmol/L glucose (P<0.05).10 mmol/L sucrose could increase significantly the nucleus maturation rates (P<0.05),and there was no significant difference of the blastocyst formation rates after IVF between 0 and 10 mmol/L sucrose (P>0.05). Furthermore, the nucleus maturation rates,IVF cleavage rates and blastocyst formation rates of yak oocytes which pretreated with 10 mmol/L glucose were the highest in these groups, and were higher than 0 mmol/L glucose (P<0.05). It manifested that the appropriate concentration of sugar could improve the quality of yak oocytes and embryos in vitro developmental competence, so it influenced in vitro development of yak oocytes indirectly. 相似文献
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Nguyen Viet LINH Kazuhiro KIKUCHI Michiko NAKAI Fuminori TANIHARA Junko NOGUCHI Hiroyuki KANEKO Thanh Quang DANG-NGUYEN Nguyen Thi MEN Nguyen VAN HANH Tamas SOMFAI Bui Xuan NGUYEN Takashi NAGAI Noboru MANABE 《The Journal of reproduction and development》2013,59(6):549-556
Mitochondria are reported to be critical in in vitro maturation of
oocytes and subsequent embryo development after fertilization, but their contribution
for fertilization has not been investigated in detail. In the present study, we
investigate the contribution of mitochondria to fertilization using reconstructed
porcine oocytes by fusion of ooplasmic fragments produced by serial centrifugations
(centri-fusion). Firstly, we evaluated the characteristics of ooplasmic fragments.
Three types of fragments were obtained by centrifugation of porcine oocytes matured
in vitro for 46 h: brownish (B), transparent (T) and large (L)
fragments containing both B and T parts in a fragment. The production efficiencies of
these types of fragments were 71.7, 91.0 and 17.8 fragments/100 oocytes,
respectively. In experiments, L fragments were excluded because they contained both
brownish and transparent components that were apparently intermediate between B and T
fragments. Observations by confocal microscopy after staining with MitoTracker Red
CMXRos® and transmission electron microscopy revealed highly condensed
active mitochondria in B fragments in contrast to T fragments that contained only
sparse organelles. We reconstructed oocytes by fusion of a karyoplast and two
cytoplasts from B and T fragments (B and T oocytes, respectively). The B oocytes
showed higher sperm penetration (95.8%) and male pronuclear formation rates (94.2%)
by in vitro fertilization than T oocytes (66.7% and 50.0%,
respectively). These results suggest that the active mitochondria in oocytes may be
related to their ability for fertilization. 相似文献
19.
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
20.
G.R. Holyoak S. Wang G. Liu T.J. Bunch R.C. Evans T.D. Bunch 《Journal of veterinary pharmacology and therapeutics》1998,21(2):92-98
Ceftiofur sodium is a third-generation cephalosporin antibiotic with broad spectrum bactericidal activity against Gram-positive and Gram-negative bacteria including the beta-lactamase producing strains. In this study, we use in vitro techniques to examine the effects of low and high levels of ceftiofur sodium on the development of bovine oocytes/embryos during oocyte maturation, oocyte fertilization and embryo culture. A total of 8590 oocytes was used in six independent experiments, each in a randomized complete block design. Each replication within each experiment consisted of oocytes from the same abattoir collection of ovaries. There was no difference in embryo development when oocytes were exposed to ceftiofur sodium during oocyte maturation or fertilization at low (10 and 50 μg/mL) or high (100 and 200 μg/mL) concentrations. However, when fertilized oocytes were exposed to concentrations 50 μg/mL during culture, ceftiofur sodium significantly retarded embryo development (e.g. the numbers of ova developing to the morula and blastocyst stages were reduced, and a large proportion of embryos were blocked at the 8-cell stage). We conclude that ceftiofur sodium does not appear to have detrimental effects on oocyte maturation and fertilization. However, long term exposure to high dosages of ceftiofur sodium during post-fertilization culture adversely affects embryo development in vitro. 相似文献