Objective:To evaluate changes of feline(Felis catus)oocytes proteins during in vitro maturation by using the proteomic approach.Methods:Immature oocytes(germinal vesicle)isolated from female cats were cultured and col...Objective:To evaluate changes of feline(Felis catus)oocytes proteins during in vitro maturation by using the proteomic approach.Methods:Immature oocytes(germinal vesicle)isolated from female cats were cultured and collected at 0 h and 24 h.After collection,oocytes were investigated into immature(germinal vesicle)and mature(metaphaseⅡ)stages.The qualitative profiles of the proteins at the immature and mature stages were determined by one-dimensional electrophoresis and liquid chromatography-mass spectrometry.Results:Our data revealed that following 24 h in vitro maturation the maturation rate(metaphaseⅡstage)was 58.7%.Eighty-one of the 260 proteins analyzed were differentially expressed between the germinal vesicle stage and the metaphaseⅡ-arrest stage.Proteomic analysis of germinal vesicle and metaphaseⅡoocytes showed abundant expression of proteins involved in transportation(10%),indicating that this was a major characteristic of germinal vesicle oocytes.Similarly,analysis of the proteome of metaphaseⅡoocytes indicated that cell cycle proteins were overexpressed.Interestingly,proteins involved in DNA repair and apoptosis were only expressed in germinal vesicle oocytes and proteins involved in fertilization were only expressed in metaphaseⅡoocytes.Conclusions:The overexpression of certain proteins in germinal vesicle and metaphaseⅡis necessary for oocyte development and maturation.Our findings provide a valuable resource for further investigations into protein expression in oocytes at different developmental stages.展开更多
[Objective]The aim was to explore optimization of system of oocytes in-vitro culture of young animals. [Method]Effects of EGF （ epidermal growth factor） and β-mercaptoethanol in maturation media on fertilization,cl...[Objective]The aim was to explore optimization of system of oocytes in-vitro culture of young animals. [Method]Effects of EGF （ epidermal growth factor） and β-mercaptoethanol in maturation media on fertilization,cleavage and blastaea were researched,which were then compared with those of adult sheep. [Result]EGF in different concentrations had little effects on rates of cleavage and blastaea （ P 〉0.05） and β-mer-captoethanol in different concentrations would improve blastaea rate of oocytes,for example,100 μmol/L of β-mercaptoethanol has significant effects on balstaea rate （ P 〈0.05） ,but has little effects on cleavage rate （ P 〉0. 05） . In addition,rates of cleavage and balstaea of oocytes in lamb were both lower than those of adult sheep （ P 〈0.05） ; fertilization rate of oocytes in lamb （ P 〈0.05） ,which differed little with that of adult sheep （ P 〉0.05） ,could be significantly enhanced by 100 μmol/L of β-mercaptoethanol. Furthermore,polyspermy rate was higher than that of adult sheep without β-mercaptoethanol （ P 〈0.05） ; the rate was of little differences with that of adult sheep with 100 μmol/L of β-mercaptoethanol （ P 〉 0.05） ; unfertilization rate （ 20%） in media without β-mercaptoethanol was a little higher （ P 〉0.05） than those of adult sheep （ 12.3%） and those in media with β-mercaptoethanol （ 13.5%） . [Conclusion]Developmental capacity of oocytes and fertilization rate could be improved by 100 μmol/L of β-mercaptoethanol with polyspermy rate reduced,but developmental capacity of lamb was significantly lower than that of adult sheep.展开更多
Objective To investigate the viability and maturation of frozen-thawed human immature oocytes exposed to different temperature of vitrification and warming solutions. Methods The immature oocytes in germinal vesicle ...Objective To investigate the viability and maturation of frozen-thawed human immature oocytes exposed to different temperature of vitrification and warming solutions. Methods The immature oocytes in germinal vesicle （GV） and matephase I （MI ） stages were collected from our ICSI patients and exposed to different temperature of vitrification and warming solutions before frozen in the freezing/thawing procedures. The different temperature groups were as follows： Group A, equilibration solution at 37℃, vitrification solution at room temperature, warming solution at 37℃; Group B, both vitrification and warming solution at room temperature; Group C, both vitrification and warming solutions at 37℃; Group D, the frozen-thawed oocytes and the fresh oocytes were cultured for in vitro maturation. The survival rate and maturation rate were compared among groups. The oocytes were examined using immunofluorescent stainingand confocal microscopy to check the spindle configuration and chromosome arrangement. Results The survival rates and MII rates of GV stage oocytes in groups A, B, C were 100%（15/15）, 81.3%（13/16）, 68.8%（11/16） and 33.3%（2/6）, 83.3%（10/12）,72.7%（8/11）, respectively. The survival rate of group C was significantly lower than that of the control （P〈0.05）. The normal spindle and chromosome configuration were only observed in group B, with the rates of 20% （2/20） and 10% （1/10）, respectively. The survival rates of MI stage in groups A, B, C were 71.4%（10/14）, 100% （12/12） and 83.3%（10/12）, no significantly difference from that of the contro1（100%, 14/14）. The MII rates of MI stage in groups A, B and C were 0%（0/14）, 66.7%（8/12） and 80% （8/10）, respectively. The MⅡ rate in group A was significantly lower than that in other groups （P〈0.01）. Only one oocyte in group C was found with normal spindle and chromosome configurations. Conclusion The appropriate operation temperature of vitrification and warming solutions can improve the outcomes of the vitrified-thawed human immature oocytes.展开更多
Human immature oocytes can be matured in vitro following culture. In vitro maturation(IVM) refers to maturation in culture of immature oocytes at different stages that may or may not have been exposed to short courses...Human immature oocytes can be matured in vitro following culture. In vitro maturation(IVM) refers to maturation in culture of immature oocytes at different stages that may or may not have been exposed to short courses of gonadotropins. The source of immature oocytes is an important feature for the subsequent embryonic development and pregnancy, as well healthy live births. IVM is an efficient treatment that has already achieved significant outcomes in terms of acceptable pregnancies and implantation rates and resulted in the births of several thousands of healthy babies. As the development of IVM treatment continues, an attractive possibility for improving the already successful outcome is to combine a natural cycle in vitro fertilization(IVF) treatment with an immature oocyte retrieval followed by IVM of those immature oocytes.If the treatment processes can be simplified with immature oocyte retrieval, different types of infertile women may be able to take advantage of these treatments. Although IVM treatment is still considered experimental by the society, it is time to reconsider the IVM technological evolution. Mild stimulation IVF combined with IVM treatment represents a viable alternative to the standard treatment, and as data accumulate over time, it may prove to be an optimal first-line treatment approach.展开更多
The technique of human in vitro fertilization and embryo transfer (IVF-ET or IVF) has been establishedt. But, at present the IVF mainly benefits the patients who are responsive to superovulation. Donation of oocytes f...The technique of human in vitro fertilization and embryo transfer (IVF-ET or IVF) has been establishedt. But, at present the IVF mainly benefits the patients who are responsive to superovulation. Donation of oocytes from healthy donors is needed in women with premature menopause, or ovarian agenesis, or dysgenesis. Many young women request展开更多
The present work was designed to examine the effect of the presence or absence of cumulus cells on the efficiency of vitrification of immature cattle oocytes. In our experiment, we had two groups: group 1, immature ca...The present work was designed to examine the effect of the presence or absence of cumulus cells on the efficiency of vitrification of immature cattle oocytes. In our experiment, we had two groups: group 1, immature cattle oocytes with cumulus cells and group 2, immature cattle oocytes without cumulus cells. The two groups underwent vitrification using 20% ethylene glycol and 20% DMSO, and then thawed, and in vitro matured in TCM-199 medium and examined after 22 hours for assessment of nuclear maturation. Higher survival rate (p < 0.05) after thawing was observed in group 1 (84.6%) than group 2 (57.8%). After in-vitro maturation, the rate of MII oocytes was significantly higher (p < 0.05) in group 1 (74.4%) than group 2 (47.7%). In conclusion, the cumulus cells are very important in increasing the survivability and developmental rate of vitrified-thawed immature cattle oocytes.展开更多
Ultrasound-guided transvaginal oocyte retrieval （TVOR） has become the gold standard for couples undergoing in-vitro fertilization （IVF）. Despite a relatively low complication rate following the procedure, here we ...Ultrasound-guided transvaginal oocyte retrieval （TVOR） has become the gold standard for couples undergoing in-vitro fertilization （IVF）. Despite a relatively low complication rate following the procedure, here we report a rare case of a ruptured ovarian abscess presenting late after ultrasound-guided TVOR in a 32-year-old woman with ovarian endometriomata. Prompt intervention and proper choices of treatment led to rapid patient recovery with no long-term sequelae. Rupture of ovarian abscess needs to be included in the differential diagnosis of a patient presenting with abdominal pain following TVOR.展开更多
Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles i...Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration. Results: A total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P < 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term. Conclusion: It appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.展开更多
Parthenogenesis is a form of asexual reproduction found in females, where growth and development of embryos occurs without fertilization by a male. Parthenogenesis occurs naturally in aphids, Daphnia, rotifers, nemato...Parthenogenesis is a form of asexual reproduction found in females, where growth and development of embryos occurs without fertilization by a male. Parthenogenesis occurs naturally in aphids, Daphnia, rotifers, nematodes and some other invertebrates but can also be induced efficiently in mammalian oocytes by providing appropriate stimuli invitro. Recently, parthenogenesis has attracted wide attention because of the role of activated oocytes in the field of research that have been described such as intra cytoplasmic sperm injection, cloning by nuclear transfer, somatic cell cloning, investigating culture conditions etc. & potential for deriving pluripotent stem cell lines and their differentiation into various cell lines that can be utilized for various tissue engineering applications. The parthenogenetically activated oocytes possess maternal genome and can developed in to either haploid, diploid or polyploidy embryos with the help of it we can analyze the possible role of all the genes involved in imprinting processes as well as the role the paternal genome plays during early embryo development by comparing them with fertilized embryos. Several methods are able to induce parthenogenetic activation through the elevation of cytoplasmic free calcium in oocytes. But one common, universal method or activation agents has not been developed for all species because the process is highly specific for each species. Therefore, activation step for each species need to be optimized accordingly. This review describes the general method of activation of mammalian oocytes and their genomic imprinting analysis.展开更多
Polycystic ovary syndrome(PCOS) is the most common cause of anovulatory infertility.If PCOS infertile women fail to conceive after conventional induction of ovulation,the assisted reproductive therapy is an alternativ...Polycystic ovary syndrome(PCOS) is the most common cause of anovulatory infertility.If PCOS infertile women fail to conceive after conventional induction of ovulation,the assisted reproductive therapy is an alternative method for pregnancy.In-vitro maturation is an efficient,more economical and simple method without ovarian hyperstimulation syndrome complication。展开更多
基金supported by a grant from the Royal Golden Jubilee,RGJ,Thailand(Grant number:PHD/0350/2551).
文摘Objective:To evaluate changes of feline(Felis catus)oocytes proteins during in vitro maturation by using the proteomic approach.Methods:Immature oocytes(germinal vesicle)isolated from female cats were cultured and collected at 0 h and 24 h.After collection,oocytes were investigated into immature(germinal vesicle)and mature(metaphaseⅡ)stages.The qualitative profiles of the proteins at the immature and mature stages were determined by one-dimensional electrophoresis and liquid chromatography-mass spectrometry.Results:Our data revealed that following 24 h in vitro maturation the maturation rate(metaphaseⅡstage)was 58.7%.Eighty-one of the 260 proteins analyzed were differentially expressed between the germinal vesicle stage and the metaphaseⅡ-arrest stage.Proteomic analysis of germinal vesicle and metaphaseⅡoocytes showed abundant expression of proteins involved in transportation(10%),indicating that this was a major characteristic of germinal vesicle oocytes.Similarly,analysis of the proteome of metaphaseⅡoocytes indicated that cell cycle proteins were overexpressed.Interestingly,proteins involved in DNA repair and apoptosis were only expressed in germinal vesicle oocytes and proteins involved in fertilization were only expressed in metaphaseⅡoocytes.Conclusions:The overexpression of certain proteins in germinal vesicle and metaphaseⅡis necessary for oocyte development and maturation.Our findings provide a valuable resource for further investigations into protein expression in oocytes at different developmental stages.
基金funded by the National Natural Science Foundation of China (31160460)Doctor Fund of Xinjiang Military Groups (2010JC10)+3 种基金National Wool Industry System (nycytx-40-06)National Key Technology Research and Development Program of the Ministry of Science and Technology of China(2011BAD28B05-1-1)863 Plan (2011AA100307)Breeding Strategy of Xinjiang Group (2011BA006)
文摘[Objective]The aim was to explore optimization of system of oocytes in-vitro culture of young animals. [Method]Effects of EGF （ epidermal growth factor） and β-mercaptoethanol in maturation media on fertilization,cleavage and blastaea were researched,which were then compared with those of adult sheep. [Result]EGF in different concentrations had little effects on rates of cleavage and blastaea （ P 〉0.05） and β-mer-captoethanol in different concentrations would improve blastaea rate of oocytes,for example,100 μmol/L of β-mercaptoethanol has significant effects on balstaea rate （ P 〈0.05） ,but has little effects on cleavage rate （ P 〉0. 05） . In addition,rates of cleavage and balstaea of oocytes in lamb were both lower than those of adult sheep （ P 〈0.05） ; fertilization rate of oocytes in lamb （ P 〈0.05） ,which differed little with that of adult sheep （ P 〉0.05） ,could be significantly enhanced by 100 μmol/L of β-mercaptoethanol. Furthermore,polyspermy rate was higher than that of adult sheep without β-mercaptoethanol （ P 〈0.05） ; the rate was of little differences with that of adult sheep with 100 μmol/L of β-mercaptoethanol （ P 〉 0.05） ; unfertilization rate （ 20%） in media without β-mercaptoethanol was a little higher （ P 〉0.05） than those of adult sheep （ 12.3%） and those in media with β-mercaptoethanol （ 13.5%） . [Conclusion]Developmental capacity of oocytes and fertilization rate could be improved by 100 μmol/L of β-mercaptoethanol with polyspermy rate reduced,but developmental capacity of lamb was significantly lower than that of adult sheep.
文摘Objective To investigate the viability and maturation of frozen-thawed human immature oocytes exposed to different temperature of vitrification and warming solutions. Methods The immature oocytes in germinal vesicle （GV） and matephase I （MI ） stages were collected from our ICSI patients and exposed to different temperature of vitrification and warming solutions before frozen in the freezing/thawing procedures. The different temperature groups were as follows： Group A, equilibration solution at 37℃, vitrification solution at room temperature, warming solution at 37℃; Group B, both vitrification and warming solution at room temperature; Group C, both vitrification and warming solutions at 37℃; Group D, the frozen-thawed oocytes and the fresh oocytes were cultured for in vitro maturation. The survival rate and maturation rate were compared among groups. The oocytes were examined using immunofluorescent stainingand confocal microscopy to check the spindle configuration and chromosome arrangement. Results The survival rates and MII rates of GV stage oocytes in groups A, B, C were 100%（15/15）, 81.3%（13/16）, 68.8%（11/16） and 33.3%（2/6）, 83.3%（10/12）,72.7%（8/11）, respectively. The survival rate of group C was significantly lower than that of the control （P〈0.05）. The normal spindle and chromosome configuration were only observed in group B, with the rates of 20% （2/20） and 10% （1/10）, respectively. The survival rates of MI stage in groups A, B, C were 71.4%（10/14）, 100% （12/12） and 83.3%（10/12）, no significantly difference from that of the contro1（100%, 14/14）. The MII rates of MI stage in groups A, B and C were 0%（0/14）, 66.7%（8/12） and 80% （8/10）, respectively. The MⅡ rate in group A was significantly lower than that in other groups （P〈0.01）. Only one oocyte in group C was found with normal spindle and chromosome configurations. Conclusion The appropriate operation temperature of vitrification and warming solutions can improve the outcomes of the vitrified-thawed human immature oocytes.
文摘Human immature oocytes can be matured in vitro following culture. In vitro maturation(IVM) refers to maturation in culture of immature oocytes at different stages that may or may not have been exposed to short courses of gonadotropins. The source of immature oocytes is an important feature for the subsequent embryonic development and pregnancy, as well healthy live births. IVM is an efficient treatment that has already achieved significant outcomes in terms of acceptable pregnancies and implantation rates and resulted in the births of several thousands of healthy babies. As the development of IVM treatment continues, an attractive possibility for improving the already successful outcome is to combine a natural cycle in vitro fertilization(IVF) treatment with an immature oocyte retrieval followed by IVM of those immature oocytes.If the treatment processes can be simplified with immature oocyte retrieval, different types of infertile women may be able to take advantage of these treatments. Although IVM treatment is still considered experimental by the society, it is time to reconsider the IVM technological evolution. Mild stimulation IVF combined with IVM treatment represents a viable alternative to the standard treatment, and as data accumulate over time, it may prove to be an optimal first-line treatment approach.
基金a grant from the Ministry of Public Health, China
文摘The technique of human in vitro fertilization and embryo transfer (IVF-ET or IVF) has been establishedt. But, at present the IVF mainly benefits the patients who are responsive to superovulation. Donation of oocytes from healthy donors is needed in women with premature menopause, or ovarian agenesis, or dysgenesis. Many young women request
文摘The present work was designed to examine the effect of the presence or absence of cumulus cells on the efficiency of vitrification of immature cattle oocytes. In our experiment, we had two groups: group 1, immature cattle oocytes with cumulus cells and group 2, immature cattle oocytes without cumulus cells. The two groups underwent vitrification using 20% ethylene glycol and 20% DMSO, and then thawed, and in vitro matured in TCM-199 medium and examined after 22 hours for assessment of nuclear maturation. Higher survival rate (p < 0.05) after thawing was observed in group 1 (84.6%) than group 2 (57.8%). After in-vitro maturation, the rate of MII oocytes was significantly higher (p < 0.05) in group 1 (74.4%) than group 2 (47.7%). In conclusion, the cumulus cells are very important in increasing the survivability and developmental rate of vitrified-thawed immature cattle oocytes.
文摘Ultrasound-guided transvaginal oocyte retrieval （TVOR） has become the gold standard for couples undergoing in-vitro fertilization （IVF）. Despite a relatively low complication rate following the procedure, here we report a rare case of a ruptured ovarian abscess presenting late after ultrasound-guided TVOR in a 32-year-old woman with ovarian endometriomata. Prompt intervention and proper choices of treatment led to rapid patient recovery with no long-term sequelae. Rupture of ovarian abscess needs to be included in the differential diagnosis of a patient presenting with abdominal pain following TVOR.
文摘Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration. Results: A total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P < 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term. Conclusion: It appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.
文摘Parthenogenesis is a form of asexual reproduction found in females, where growth and development of embryos occurs without fertilization by a male. Parthenogenesis occurs naturally in aphids, Daphnia, rotifers, nematodes and some other invertebrates but can also be induced efficiently in mammalian oocytes by providing appropriate stimuli invitro. Recently, parthenogenesis has attracted wide attention because of the role of activated oocytes in the field of research that have been described such as intra cytoplasmic sperm injection, cloning by nuclear transfer, somatic cell cloning, investigating culture conditions etc. & potential for deriving pluripotent stem cell lines and their differentiation into various cell lines that can be utilized for various tissue engineering applications. The parthenogenetically activated oocytes possess maternal genome and can developed in to either haploid, diploid or polyploidy embryos with the help of it we can analyze the possible role of all the genes involved in imprinting processes as well as the role the paternal genome plays during early embryo development by comparing them with fertilized embryos. Several methods are able to induce parthenogenetic activation through the elevation of cytoplasmic free calcium in oocytes. But one common, universal method or activation agents has not been developed for all species because the process is highly specific for each species. Therefore, activation step for each species need to be optimized accordingly. This review describes the general method of activation of mammalian oocytes and their genomic imprinting analysis.
文摘Polycystic ovary syndrome(PCOS) is the most common cause of anovulatory infertility.If PCOS infertile women fail to conceive after conventional induction of ovulation,the assisted reproductive therapy is an alternative method for pregnancy.In-vitro maturation is an efficient,more economical and simple method without ovarian hyperstimulation syndrome complication。
