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1.
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
2.
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
3.
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
4.
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
5.
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
6.
Yamato MIZOBE Saori KURINO Yoshiaki SATA Hironori MORI Mitsutoshi YOSHIDA Kazuchika MIYOSHI 《Animal Science Journal》2010,81(4):453-460
Whether high osmolarity of a culture medium at the early culture stage affects the development of pig oocytes and miniature pig somatic cell nuclear transfer (SCNT) embryos activated by ultrasound was examined. When oocytes were cultured in modified porcine zygote medium‐3 (mPZM‐3) with increased NaCl to 138 mmol/L (mPZM‐3+NaCl; 326 mOsm) or 50 mmol/L sucrose (mPZM‐3+sucrose; 318 mOsm) for the first 2 days and then cultured in normal mPZM‐3 (273 mOsm) for 5 days, the cleavage and blastocyst formation rates were significantly (P < 0.05) higher than those of oocytes cultured in mPZM‐3 for 7 days. The cleavage and blastocyst formation rates of SCNT embryos cultured in mPZM‐3+NaCl for the first 2 days and then cultured in mPZM‐3 for 5 days were also significantly (P < 0.05) higher than those of embryos cultured in mPZM‐3 for 7 days. These results showed that the high osmolarity of a culture medium induced by increasing NaCl concentration during the first 2 days improves the development of pig oocytes and miniature pig SCNT embryos activated by ultrasound. 相似文献
7.
Jungmin Son Don Buddika Oshadi Malaweera Eunsong Lee Sangtae Shin Jongki Cho 《Journal of veterinary science (Suw?n-si, Korea)》2013,14(3):315-321
This study was conducted to establish an in vitro maturation (IVM) system by selection of efficient porcine serum during porcine in vitro production. To investigate the efficient porcine serum (PS), different types of PS [newborn pig serum, prepubertal gilt serum (PGS), estrus sow serum, and pregnancy sow serum] were used to supplement IVM media with or without gonadotrophin (GTH) and development rates of parthenogenetic activation (PA) and in vitro fertilization (IVF) embryos were then compared. The maturation rates of the PGS group was significantly higher when GTH was not added. Additionally, during development of PA embryos without GTH, the PGS group showed significantly higher cleavage and blastocyst formation rates. Moreover, the cleavage rates of IVF embryos were significantly higher in the PGS group, with no significant differences in the blastocyst formation. However, when GTH was supplemented into the IVM media, there were no significant differences among the four groups in the cleavage rates, development rates of the blastocyst, and cell number of the blastocyst after PA and IVF. In conclusion, PGS is an efficient macromolecule in porcine IVM, and GTH supplementation of the IVM media is beneficial when PS is used as macromolecule, regardless of its origin. 相似文献
8.
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
9.
This study examined effects on the developmental competence of pig oocytes after somatic cell nuclear transfer (SCNT) or parthenogenetic activation (PA) of : 1) co-culturing of oocytes with follicular shell pieces (FSP) during in vitro maturation (IVM); 2) different durations of maturation; and 3) defined maturation medium supplemented with polyvinyl alcohol (PVA; control), pig follicular fluid (pFF), cysteamine (CYS), or β-mercaptoethanol (β-ME). The proportion of metaphase II oocytes was increased (p < 0.05) by co-culturing with FSP compared to control oocytes (98% vs. 94%). However, blastocyst formation after SCNT was not improved by FSP coculture (9% vs. 12%). Nuclear maturation of oocytes matured for 39 or 42 h was higher (p < 0.05) than that of oocytes matured for 36 h (95-96% vs. 79%). Cleavage (83%) and blastocyst formation (26%) were significantly higher (p < 0.05) in oocytes matured for 42 h than in other groups. Supplementation of a defined maturation medium with 100 µM CYS or 100 µM β-ME showed no stimulatory effect on oocyte maturation, embryo cleavage, or blastocyst formation after PA. β-ME treatment during IVM decreased embryo cleavage after SCNT compared to pFF or PVA treatments, but no significant difference was found in blastocyst formation (7-16%) among the four treatment groups. The results indicated that maturation of oocytes for 42 h was beneficial for the development of SCNT embryos. Furthermore, the defined maturation system used in this study could support in vitro development of PA or SCNT embryos. 相似文献
10.
为探讨表皮生长因子(epidermal growth factor,EGF)的添加浓度及脱卵丘细胞时间对猪卵母细胞体外成熟及孤雌胚胎体外发育的影响.试验通过在体外成熟液中添加不同浓度(0、10、15、20、30、40 ng/mL)的EGF来研究其对培养44 h的卵母细胞成熟率以及孤雌胚胎发育的影响;在培养开始后的不同时间(18、24、38、44 h)进行脱卵丘细胞处理来研究不同时间脱卵丘处理对培养44 h的卵母细胞成熟率以及孤雌胚胎发育的影响.结果表明,成熟培养基中添加10 ng/mL EGF能显著提高卵母细胞的卵裂率和囊胚率(P <0.05).共培组和独培组卵母细胞培养18 h后脱卵丘细胞成熟率均低于44 h,但差异不显著(P >0.05);共培组卵母细胞培养18 h后脱卵丘细胞的卵裂率和囊胚率显著高于培养44 h(P <0.05);独培组卵母细胞培养18 h后脱卵丘细胞的卵裂率与44 h无显著差异(P >0.05),但囊胚率显著高于培养44 h后脱卵丘细胞(P <0.05).添加10 ng/mL EGF对猪卵母细胞体外成熟及孤雌胚胎体外发育较好;卵母细胞培养18 h后脱卵丘细胞可提高孤雌胚胎早期发育能力. 相似文献
11.
S.K. Das Aditya K. SharmaSushil K. Mohapatra Vishu BhatiaA. Chatterjee A.K. Mohanty 《Livestock Science》2013,152(1):88-93
The purpose of this study was to determine the effect of oviduct specific proteins as a media supplement for in vitro embryo development in cattle. The proteins were extracted from oviducts of cows and precipitated by ammonium sulfate (30%, 40%, 50% and 60%) followed by dialysis in 50 mM Tris–HCl (pH 7.0) buffer. The dialyzed proteins were fractionated into acidic, basic and neutral fractions using SP sephadex cation exchange and DEAE sephadex anion exchange column chromatography respectively. Cow oviduct specific proteins (cOSPs) constituting all the extracted proteins were used as media supplement in three different concentrations (10, 50 and 100 μg/ml) for in vitro maturation, fertilization and culture (IVMFC) of cow oocytes. Acidic, basic and neutral (unbound) fractions were also used as media supplement in three different concentrations (10, 30 and 50 μg/ml) for IVMFC. Cumulus oocytes complexes were collected from slaughterhouse ovaries, washed thoroughly and cultured in maturation media for 24 h in 5% CO2 at 38.5 °C with maximum humidity. In vitro matured oocytes were co-incubated with in vitro capacitated sperm in Fert-BO media at 38.5 °C for 18 h in 5% CO2. The fertilized oocytes were washed and cultured in embryo development media for cleavage. After 40–42 h cleavage was observed and embryos were put in the replacement media for further development. The cleavage rates (%) for cOSPs were observed as 68.24±2.46, 69.28±2.05, 61.77±0.93 and 42.62±1.31 at concentrations of 0, 10, 50 and 100 μg/ml respectively. Rates of blastocyst stage development were 14.49±3.61, 21.17±2.77, 14.66±1.06 and 11.98±1.84. These results indicate that addition of cOSP at10 μg/ml increased blastocyst formation as compared to other concentrations (0, 50 and 100 μg/ml). Although acidic, basic and neutral fractions seemed to have no major effect on cleavage rate, but both acidic and neutral fraction of oviduct specific proteins improved the cleavage rate at 30 μg/ml concentration and basic fraction improved the blastocyst formation at 10 μg/ml concentration. 相似文献
12.
Yan Jun HUAN Jiang ZHU Bing Teng XIE Jian Yu WANG Shi Chao LIU Yang ZHOU Qing Ran KONG Hong Bin HE Zhong Hua LIU 《The Journal of reproduction and development》2013,59(5):442-449
The efficiency of cloning by somatic cell nuclear transfer (SCNT) has remained low.
In most cloned embryos, epigenetic reprogramming is incomplete, and usually the
genome is hypermethylated. The DNA methylation inhibitor 5-aza-2’-deoxycytidine
(5-aza-dC) could improve the developmental competence of cow, pig, cat and human SCNT
embryos in previous studies. However, the parameters of 5-aza-dC treatment among
species are different, and whether 5-aza-dC could enhance the developmental
competence of porcine cloned embryos has still not been well studied. Therefore, in
this study, we treated porcine fetal fibroblasts (PFF) that then were used as donor
nuclei for nuclear transfer or fibroblast-derived reconstructed embryos with
5-aza-dC, and the concentration- and time-dependent effects of 5-aza-dC on porcine
cloned embryos were investigated by assessing pseudo-pronucleus formation,
developmental potential and pluripotent gene expression of these reconstructed
embryos. Our results showed that 5-aza-dC significantly reduced the DNA methylation
level in PFF (0 nM vs. 10 nM vs. 25 nM
vs. 50 nM, 58.70% vs. 37.37%
vs. 45.43% vs. 39.53%, P<0.05), but did not
improve the blastocyst rate of cloned embryos derived from these cells. Treating
cloned embryos with 25 nM 5-aza-dC for 24 h significantly enhanced the blastocyst
rate compared with that of the untreated group. Furthermore, treating cloned embryos,
but not donor cells, significantly promoted pseudo-pronucleus formation at 4 h post
activation (51% for cloned embryos treated, 34% for donor cells treated and 36% for
control, respectively, P<0.05) and enhanced the expression levels of pluripotent
genes (Oct4, Nanog and Sox2) up to
those of in vitro fertilized embryos during embryo development. In
conclusion, treating cloned embryos, but not donor cells, with 5-aza-dC enhanced the
developmental competence of porcine cloned embryos by promotion of pseudo-pronucleus
formation and improvement of pluripotent gene expression. 相似文献
13.
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
14.
A Mukherjee D Kumar KP Singh MS Chauhan SK Singla P Palta RS Manik 《Reproduction in domestic animals》2010,45(6):1118-1121
Comet assay was used in the present study to examine DNA damage to buffalo oocytes and embryos during in vitro culture. Embryos were produced in vitro from oocytes obtained from slaughterhouse ovaries in presence of cysteamine (IVM and IVC media supplemented with 50 and 100 μm , respectively) or in its absence (controls). Compared to controls, cysteamine supplementation increased (p < 0.01) cleavage rate and proportion of oocytes that developed to 8‐ to 16‐cell stage. The incidence of DNA damage was lower (p < 0.01) in cysteamine group than that in controls at 8‐ to 16‐ (19.3 ± 4.24 vs 72.0 ± 5.22%) but not in 2‐cell stage embryos (11.7 ± 5.63 vs 20.8 ± 5.49%) or in mature oocytes (5.3 ± 3.43 vs 10.3 ± 4.73%). The tail length, which indicates magnitude of DNA damage, was shorter (p < 0.01) in cysteamine group than in controls in mature oocytes (25.5 ± 0.5 vs 36.0 ± 0.71 pixels) and 8‐ to 16‐cell stage (49.2 ± 1.64 vs 152.7 ± 1.28 pixels) but not in 2‐cell stage embryos (36.3 ± 1.54 vs 36.4 ± 0.75 pixels). Also, exposure of oocytes/embryos to UV radiation or H2O2 caused extensive DNA damage. In conclusion, these results suggest that oocytes/embryos suffer from DNA damage during progress of in vitro culture, which can be partly ameliorated by cysteamine supplementation of culture media. 相似文献
15.
Charley-Lea POLLARD Zamira GIBB Azelle HAWDON Aleona SWEGEN Christopher G. GRUPEN 《The Journal of reproduction and development》2021,67(5):319
In vitro maturation (IVM) is an important reproductive technology used to produce embryos in vitro. However, the developmental potential of oocytes sourced for IVM is markedly lower than those matured in vivo. Previously, NAD+-elevating treatments have improved oocyte quality and embryo development in cattle and mice, suggesting that NAD+ is important during oocyte maturation. The aim of this study was to examine the effects of nicotinic acid (NA), nicotinamide (NAM) and nicotinamide mononucleotide (NMN) on oocyte maturation and subsequent embryo development. Porcine oocytes from small antral follicles were matured for 44 h in a defined maturation medium supplemented with NA, NAM and resveratrol or NMN. Mature oocytes were artificially activated and presumptive zygotes cultured for 7 days. Additionally, oocytes were matured without treatment then cultured for 7 days with NMN. Supplementing the IVM medium with NA improved maturation and blastocyst formation while NAM supplementation improved cleavage rates compared with untreated controls. Supplementing the IVM or embryo culture media with NMN had no effect on maturation or embryo development. The results show that supplementing the maturation medium with NA and NAM improved maturation and developmental potential of porcine oocytes. 相似文献
16.
Shun TAKEO Hiroya GOTO Takehito KUWAYAMA Yasunori MONJI Hisataka IWATA 《The Journal of reproduction and development》2013,59(2):174-179
Age-associated deterioration in both the quality and quantity of mitochondria occurs in
older women. The main aim of this study was to examine the effect of age on mitochondrial
DNA copy number (mtDNA number) in early developmental stage bovine embryos as well as the
dynamics of mtDNA number during early embryo development. Real-time PCR was used to
determine mtDNA number. In vitro-produced embryos 48 h after insemination
derived from Japanese black cows, ranging in age from 25 to 209 months were categorized
based on their cleavage status. There was an overall negative relationship between the age
of the cow and cleavage status, to the extent that the ratio of embryos cleaved over the
4-cell stage was greater in younger cows. The mtDNA number did not differ among the
cleaved status of embryos. In the next experiment, oocytes collected from each donor cow
were divided into 2 groups containing 10 oocytes each, in order to compare the mtDNA
number of mature oocytes and early developmental stage embryos within individuals. Upon
comparing the mtDNA number between oocytes at the M2 stage and early developmental stage
48 h post insemination, mtDNA number was found to decrease in most cows, but was found to
increase in some cows. In conclusion, age affects the cleaving ability of oocytes, and
very old cows (> 180 months) tend to have lower mtDNA numbers in their oocytes. The
change in mtDNA number during early development varied among individual cows, although
overall, it showed a tendency to decrease. 相似文献
17.
Chommanart THONGKITTIDILOK Theerawat THARASANIT Nucharin SONGSASEN Thanida SANANMUANG Sirirak BUARPUNG Mongkol TECHAKUMPHU 《The Journal of reproduction and development》2015,61(4):269-276
This study examined the influence of EGF on the expression of EGF receptors (EGFR) and developmental competence of embryos cultured individually versus those cultured in groups. Cat oocytes were in vitro matured and fertilized (IVM/IVF), and cleaved embryos were randomly assigned to one of seven culture conditions: one group each in which embryos were subjected to group culture supplemented with or without 5 ng/ml EGF and five groups in which embryos were subjected to single-embryo culture supplemented with EGF (0, 5, 25, 50 or 100 ng/ml). Morulae, blastocysts and hatching blastocysts were assessed at days 5 and 7; post IVF, respectively, and total blastocyst cell numbers were assessed at day 7. Relative mRNA expressions of EGFR of 2–4-cell embryos, 8–16-cell embryos, morulae and blastocysts cultured in groups or singly with or without EGF supplementation were examined. OCT3/4 and Ki67 in blastocysts derived from the group
or single-embryo culture systems with or without EGF supplementation were localized. A higher rate of embryos cultured in groups developed to blastocysts than individually incubated cohorts. Although EGF increased blastocyst formation in the single-embryo culture system, EGF did not affect embryo development in group culture. Expression levels of EGFR decreased in morulae and blastocysts cultured with EGF. An increased ratio of Ki67-positive cells to the total number of cells in the blastocyst was observed in singly cultured embryos in the presence of EGF. However, EGF did not affect the expression of OCT3/4. These findings indicate that EGF enhanced developmental competence of cat embryos cultured singly by stimulating cell proliferation and modulating the EGFR expression at various developmental stages. 相似文献
18.
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