Embryo cryopreservation（CP） has became a very important part of the clinical use of in vitro fertilization. Oocyte CP offers more advantages compared with embryo freezing with regard to less ethical, legal and moral...Embryo cryopreservation（CP） has became a very important part of the clinical use of in vitro fertilization. Oocyte CP offers more advantages compared with embryo freezing with regard to less ethical, legal and moral problems. However, the efficiency of this procedure is still low, which prevents its clinical application in wide range. The aim of our paper is to review the basic principles, technical and safety aspects and current status of oocyte cryopreservation in human assisted reproduction.展开更多
Our study focused on a retrospective analysis from 2004-2011 of patients considering elective oocyte cryopreservation (OC). We investigated the psychological and social aspects related to women who electively cryopres...Our study focused on a retrospective analysis from 2004-2011 of patients considering elective oocyte cryopreservation (OC). We investigated the psychological and social aspects related to women who electively cryopreserve oocytes. Over seven years, consulted patients (n = 315) considering non-medical OC were interviewed by the staff therapists. Social, demographic, motivational impetus, psychological factors and local to national economy were analyzed in association with trends in elective OC. Patient disclosure, fertility assessment and receptivity to potential single motherhood were other aspects examined. Statistical analysis was performed with Student’s t-test, Pearson’s correlation and Chi-square analysis. Advanced technology, decreased age (<35), anual per capita income, levels of follicular stimulant hormone (FSH) and basal antral follicular count (BAFC) were demonstrated to be the most influential factors of elective OC. The mean age of elective OC patients was 38.6 ± 1.83 with nearly 80% of these patients disclosing their decisions either with family and/or friends. Clinical perception has increasingly improved the availability and efficacy of elective oocyte cryopreservation, albeit minimal publications have studied the social and epidemiological aspects of such patients. We identified these patients are often motivated by a key life event such as a birthday, are educated and professional, and often disclose their treatment to close friends and family. Understanding the psychological aspects of egg freezing patients will engender clinicians the ability to meet patients’ needs and appropriately counsel them.展开更多
Objective:To compare embryonic development,ploidy status and clinical outcomes between fresh and frozen-thawed oocytes.Methods:This retrospective cohort study evaluated 83 fertilization cycles including both fresh and...Objective:To compare embryonic development,ploidy status and clinical outcomes between fresh and frozen-thawed oocytes.Methods:This retrospective cohort study evaluated 83 fertilization cycles including both fresh and frozen oocytes from 79 patients at the HP Fertility Center of Hai Phong International Hospital of Obstetrics and Pediatrics in Vietnam.The patient underwent several ovarian stimulation cycles to accumulate a certain number of oocytes that would be vitrified.In the last oocyte retrieval,all patient’s oocytes including both frozen and fresh would be fertilized.The outcomes included the rates of oocyte survival,cleavage embryo,blastocyst,ploidy status,pregnancy,biochemical pregnancy and clinical pregnancy.Results:The oocyte survival rate after thawing was 96.5%.No statistically significant difference was found when comparing fresh and frozen oocytes regarding fertilization rate(78.1%vs.75.5%,P=0.461),usable cleavage embryo rate(86.9%vs.87.2%,P=0.916)but usable blastocyst rate was found higher statistically in the frozen oocyte group(44.4%vs.54.0%,P=0.049).The percentages of euploid,aneuploid and mosaic embryos between the fresh group and the vitrified group had no significant differences(33.8%vs.31.6%,P=0.682;51.0%vs.54.2%,P=0.569;15.2%vs.12.4%,P=0.787;respectively).The rates of pregnancy,biochemical pregnancy and clinical pregnancy had no statistical difference(68.8%vs.64.8%,P=0.764;12.5%vs.3.6%,P=0.258;37.5%vs.46.4%,P=0.565).17 Mature oocytes are the minimum to have at least one euploid embryo.Conclusions:Oocyte vitrification does not affect embryonic,genetic and clinical results.The number of mature oocytes should be considered for fertilization in some cases.展开更多
Objective To evaluate the value of oocyte cryopreservation （CP） in our clinical ICSI program. Methods Freezing procedure with medium containing 1.5 mol/L propanediol （PrOH） ＋ 0. 3 mol/L succrose and traditional ...Objective To evaluate the value of oocyte cryopreservation （CP） in our clinical ICSI program. Methods Freezing procedure with medium containing 1.5 mol/L propanediol （PrOH） ＋ 0. 3 mol/L succrose and traditional slow-freezingprotocol were used. Thawed oocytes were fertilized with ICSI （4-6 h after thawing）, and fertilization was assessed 12-16 h later. Laser assisted hatching was performed on all transferred embryos and embryo transfer was carried out 48-72 h after ICSI. Results Eighty-five eggs were thawed and survival rate of 75.3% （64/85） was obtained. Sixty-four oocytes were inseminated with ICSI, 47fertilized （47/64; 73.4%） and a cleavage rate of 85% （40/47） was obtained. Embryo transfers were performed in 18 patients, and 4 （19%） resulted in clinical pregnancies. One of the pregnancies encountered first trimester abortion. Implantation rate were 17.2% （5/29） per embryo and 5.8% （5/85） per egg thawed. In all cases, chorion biopsy was performed resulting 46 XY kariotype. Conclusion Our results provide further evidence of that although egg freezing cannot currently claim to be a routine procedure in human IVF,, there will certainly be a place for oocyte CP in reproductive medicine in the future.展开更多
Oocyte cryopreservation has recently emerged as an option for women to preserve their fertility for medical (e.g. treatable malignancy) or elective indications (e.g. advancing age). This report describes an IRB-approv...Oocyte cryopreservation has recently emerged as an option for women to preserve their fertility for medical (e.g. treatable malignancy) or elective indications (e.g. advancing age). This report describes an IRB-approved study of over 200 oocyte cryopreservation cycles at one center. Patients presenting for oocyte cryopreservation (January 2005 to 2010) were analyzed for day 3 follicle stimulating hormone (FSH), basal antral follicle count (BAFC), gonadotropin usage and the number of oocytes retrieved and cryopreserved. New patient consultations were performed on 516 women, of whom 175 (34%) proceeded to initiate a total of 233 cryopreservation cycles. Twenty-four cycles were cancelled (10%) after starting follicular stimulation due to poor ovarian response or self-withdrawal of the patients. Patients whose cycles were cancelled demonstrated a higher Day 3 FSH and a lower BAFC than patients who completed cycles (p < 0.01). In the 209 completed cycles, the most important predictors of a successful cycle included BAFC (r = 0.36), FSH (r = –0.25) and age (r = –0.18) with the mean number of oocytes cryopreserved at 13.6 ± 8.8. Information about long-term fertility preservation must reach both patients and health care providers so that more women can be educated about the benefits of proactive early physiological reproductive assessment and possible interventions available.展开更多
Background：At present,vitrification has been widely applied to humans,mice and farm animals.To improve the efficiency of vitrification in straw,bovine oocytes were used to test a new two-step vitrification method in ...Background：At present,vitrification has been widely applied to humans,mice and farm animals.To improve the efficiency of vitrification in straw,bovine oocytes were used to test a new two-step vitrification method in this study.Results：When in vitro matured oocytes were exposed to 20%ethylene glycol（EG20） for 5 min and 40%ethylene glycol（EG40） for 30 s,followed by treatment with 30%glycerol（Gly30）,Gly40 or Gly50,a volume expansion was observed in Gly30 and Gly40 but not Gly50.This indicates that the intracellular osmotic pressure after a 30 s differs between EG40 and ranged between Gly40（approximately 5.6 mol/L） and Gly50（approximately 7.0 mol/L）.Since oocytes are in EG40 just for only a short period of time（30 s） and at a lower temperature（4℃）,we hypothesize that the main function of this step in to induce dehydration.Based on these results,we omitted the EG40 step,before oocytes were pretreated in EG20 for 5 min,exposed to pre-cooled（4℃） Gly50,for 30 s,and then dipped into liquid nitrogen.After warming,81.1%of the oocytes survived,and the surviving oocytes developed into cleavage stage embryos（63.5%） or blastocysts（20.0%） after parthenogenetic activation.Conclusions：These results demonstrate that in a two-step vitrification procedure,the permeability effect in the second step is not necessary.It is possible that the second step is only required to provide adequate osmotic pressure to condense the intracellular concentration of CPAs to a level required for successful vitrification.展开更多
Is there a really need to validate oocyte vitrification technique in an ART laboratory before establishing it in daily practice? Validation of micromanipulationbased technique, in this case oocyte vitrification, is es...Is there a really need to validate oocyte vitrification technique in an ART laboratory before establishing it in daily practice? Validation of micromanipulationbased technique, in this case oocyte vitrification, is essential prior to enlarging its use to routine practice. Oocyte vitrification is a new worldwide used technique and legal recently inFrance. This micromanipulation needs to be performed by a skilled and experienced embryologist and requires an internal assessment in each ART unit before any wide use. We designed a prospective study, from September 2011 to July 2012, using sibling oocytes from women who recovered more than 12 Metaphase II oocytes. A part of freshly recovered oocytes underwent immediate ICSI while the remaining oocytes were vitrified. 87 couples undergoing ICSI were selected based on number of mature oocytes available on the recovery day after denudation. A part of fresh MII oocytes were microinjected and the others were vitrified using an open system (Cryotop?). The major criterion of interest was the number of embryo transferred/ number of Metaphase II ratio for after ICSI on fresh oocytes (42/211) versus vitrified/warmed oocytes (51/204) (p > 0.05). Secondary studied criteria were survival rate (80.5% ± 26.3%), fertilization rate (68.9 ± 33.5) and finally, cumulative pregnancy rate obtained in this study is 40.2%. One of the benefits of such practice is the limitation of embryo freezing. However, the study design delays oocytes warming cycles, due to pregnancies triggered by the transfer of fresh derived oocyte embryos and to the priority to transfer all the frozen embryos before starting oocytes warming. Moreover, no data is available about children’ health. Oocyte vitrification represents not only a change in our daily practice to improve cumulative pregnancy rate but also a promising tool to develop egg banking and donation. Clinical Trials Registration number: 209 R02.展开更多
Cryopreservation techniques for mammalian oocytes and embryos have rapidly progressed during the past two decades,emphasizing their importance in various assisted reproductive technologies.Pregnancies and live births ...Cryopreservation techniques for mammalian oocytes and embryos have rapidly progressed during the past two decades,emphasizing their importance in various assisted reproductive technologies.Pregnancies and live births resulting from cryopreserved oocytes and embryos of several species including humans have provided proof of principle and led to the adoption of cryopreservation as an integral part of clinical in vitro fertilization.Considerable progress has been achieved in the development and application of the cryopreservation of mammalian oocytes and embryos,including preservation of the reproductive potential of patients who may become infertile,establishment of cryopreserved oocyte banks,and transport of oocytes and embryos internationally.However,the success rates are still far lower than those obtained with fresh oocytes and embryos,and there are still obstacles that need to be overcome.In this review,we address the major obstacles in the development of effective cryopreservation techniques.Such knowledge may help to eliminate these hurdles by revealing which aspects need improvement.Furthermore,this information may encourage further research by cryobiologists and increase the practical use of cryopreservation as a major part of assisted reproductive technologies for both humans and animal species.展开更多
Reproduction technologies(RTs)can provide for the reliable reproduction of amphibians,as well as perpetuation of species genetic variation with the use of biobanks.In 1982,in anticipation of the biodiversity conservat...Reproduction technologies(RTs)can provide for the reliable reproduction of amphibians,as well as perpetuation of species genetic variation with the use of biobanks.In 1982,in anticipation of the biodiversity conservation crisis,major Russian institutions collaborated in a dynamic program to develop and implement RTs for the sustainable management of amphibian biodiversity.An initial primary focus was the captive breeding of threatened Russian endemic anuran and caudate species,using RTs that varied from environmental manipulation to the use of exogenous gonadotropic hormones to stimulate reproduction.These species were mostly from Palearctic or cool mountain regions,but also included a wide range of species from warm regions.Other early achievements included the successful cryopreservation of anuran spermatozoa and anuran diploid pluripotent cell nuclei,in order to store both the matrilineal and patrilineal genomes in biobanks,with their subsequent development to the blastula stage after implantation into enucleated oocytes.After the turn of the 21st Century,in support of the priorities of the Amphibian Conservation Action Plan(2007),we developed RTs for the refrigerated storage of testicular or urinary spermatozoa for days to weeks at 4℃,the cryopreservation of urinary spermatozoa using anovel cryoprotectant,the in vitro fertilisation of hormonally induced oocytes either fresh or after refrigerated ex situ or in situ storage,and the artificial insemination of salamanders with fresh spermatozoa.In this article,we describe previously unpublished techniques and techniques from obscure Russian sources.展开更多
文摘Embryo cryopreservation（CP） has became a very important part of the clinical use of in vitro fertilization. Oocyte CP offers more advantages compared with embryo freezing with regard to less ethical, legal and moral problems. However, the efficiency of this procedure is still low, which prevents its clinical application in wide range. The aim of our paper is to review the basic principles, technical and safety aspects and current status of oocyte cryopreservation in human assisted reproduction.
文摘Our study focused on a retrospective analysis from 2004-2011 of patients considering elective oocyte cryopreservation (OC). We investigated the psychological and social aspects related to women who electively cryopreserve oocytes. Over seven years, consulted patients (n = 315) considering non-medical OC were interviewed by the staff therapists. Social, demographic, motivational impetus, psychological factors and local to national economy were analyzed in association with trends in elective OC. Patient disclosure, fertility assessment and receptivity to potential single motherhood were other aspects examined. Statistical analysis was performed with Student’s t-test, Pearson’s correlation and Chi-square analysis. Advanced technology, decreased age (<35), anual per capita income, levels of follicular stimulant hormone (FSH) and basal antral follicular count (BAFC) were demonstrated to be the most influential factors of elective OC. The mean age of elective OC patients was 38.6 ± 1.83 with nearly 80% of these patients disclosing their decisions either with family and/or friends. Clinical perception has increasingly improved the availability and efficacy of elective oocyte cryopreservation, albeit minimal publications have studied the social and epidemiological aspects of such patients. We identified these patients are often motivated by a key life event such as a birthday, are educated and professional, and often disclose their treatment to close friends and family. Understanding the psychological aspects of egg freezing patients will engender clinicians the ability to meet patients’ needs and appropriately counsel them.
文摘Objective:To compare embryonic development,ploidy status and clinical outcomes between fresh and frozen-thawed oocytes.Methods:This retrospective cohort study evaluated 83 fertilization cycles including both fresh and frozen oocytes from 79 patients at the HP Fertility Center of Hai Phong International Hospital of Obstetrics and Pediatrics in Vietnam.The patient underwent several ovarian stimulation cycles to accumulate a certain number of oocytes that would be vitrified.In the last oocyte retrieval,all patient’s oocytes including both frozen and fresh would be fertilized.The outcomes included the rates of oocyte survival,cleavage embryo,blastocyst,ploidy status,pregnancy,biochemical pregnancy and clinical pregnancy.Results:The oocyte survival rate after thawing was 96.5%.No statistically significant difference was found when comparing fresh and frozen oocytes regarding fertilization rate(78.1%vs.75.5%,P=0.461),usable cleavage embryo rate(86.9%vs.87.2%,P=0.916)but usable blastocyst rate was found higher statistically in the frozen oocyte group(44.4%vs.54.0%,P=0.049).The percentages of euploid,aneuploid and mosaic embryos between the fresh group and the vitrified group had no significant differences(33.8%vs.31.6%,P=0.682;51.0%vs.54.2%,P=0.569;15.2%vs.12.4%,P=0.787;respectively).The rates of pregnancy,biochemical pregnancy and clinical pregnancy had no statistical difference(68.8%vs.64.8%,P=0.764;12.5%vs.3.6%,P=0.258;37.5%vs.46.4%,P=0.565).17 Mature oocytes are the minimum to have at least one euploid embryo.Conclusions:Oocyte vitrification does not affect embryonic,genetic and clinical results.The number of mature oocytes should be considered for fertilization in some cases.
文摘Objective To evaluate the value of oocyte cryopreservation （CP） in our clinical ICSI program. Methods Freezing procedure with medium containing 1.5 mol/L propanediol （PrOH） ＋ 0. 3 mol/L succrose and traditional slow-freezingprotocol were used. Thawed oocytes were fertilized with ICSI （4-6 h after thawing）, and fertilization was assessed 12-16 h later. Laser assisted hatching was performed on all transferred embryos and embryo transfer was carried out 48-72 h after ICSI. Results Eighty-five eggs were thawed and survival rate of 75.3% （64/85） was obtained. Sixty-four oocytes were inseminated with ICSI, 47fertilized （47/64; 73.4%） and a cleavage rate of 85% （40/47） was obtained. Embryo transfers were performed in 18 patients, and 4 （19%） resulted in clinical pregnancies. One of the pregnancies encountered first trimester abortion. Implantation rate were 17.2% （5/29） per embryo and 5.8% （5/85） per egg thawed. In all cases, chorion biopsy was performed resulting 46 XY kariotype. Conclusion Our results provide further evidence of that although egg freezing cannot currently claim to be a routine procedure in human IVF,, there will certainly be a place for oocyte CP in reproductive medicine in the future.
文摘Oocyte cryopreservation has recently emerged as an option for women to preserve their fertility for medical (e.g. treatable malignancy) or elective indications (e.g. advancing age). This report describes an IRB-approved study of over 200 oocyte cryopreservation cycles at one center. Patients presenting for oocyte cryopreservation (January 2005 to 2010) were analyzed for day 3 follicle stimulating hormone (FSH), basal antral follicle count (BAFC), gonadotropin usage and the number of oocytes retrieved and cryopreserved. New patient consultations were performed on 516 women, of whom 175 (34%) proceeded to initiate a total of 233 cryopreservation cycles. Twenty-four cycles were cancelled (10%) after starting follicular stimulation due to poor ovarian response or self-withdrawal of the patients. Patients whose cycles were cancelled demonstrated a higher Day 3 FSH and a lower BAFC than patients who completed cycles (p < 0.01). In the 209 completed cycles, the most important predictors of a successful cycle included BAFC (r = 0.36), FSH (r = –0.25) and age (r = –0.18) with the mean number of oocytes cryopreserved at 13.6 ± 8.8. Information about long-term fertility preservation must reach both patients and health care providers so that more women can be educated about the benefits of proactive early physiological reproductive assessment and possible interventions available.
基金supported by the National "863" Project Foundation of China(No.2011AA100303)the National Science and Technology Support Projects of China(No.2011BAD19B01)
文摘Background：At present,vitrification has been widely applied to humans,mice and farm animals.To improve the efficiency of vitrification in straw,bovine oocytes were used to test a new two-step vitrification method in this study.Results：When in vitro matured oocytes were exposed to 20%ethylene glycol（EG20） for 5 min and 40%ethylene glycol（EG40） for 30 s,followed by treatment with 30%glycerol（Gly30）,Gly40 or Gly50,a volume expansion was observed in Gly30 and Gly40 but not Gly50.This indicates that the intracellular osmotic pressure after a 30 s differs between EG40 and ranged between Gly40（approximately 5.6 mol/L） and Gly50（approximately 7.0 mol/L）.Since oocytes are in EG40 just for only a short period of time（30 s） and at a lower temperature（4℃）,we hypothesize that the main function of this step in to induce dehydration.Based on these results,we omitted the EG40 step,before oocytes were pretreated in EG20 for 5 min,exposed to pre-cooled（4℃） Gly50,for 30 s,and then dipped into liquid nitrogen.After warming,81.1%of the oocytes survived,and the surviving oocytes developed into cleavage stage embryos（63.5%） or blastocysts（20.0%） after parthenogenetic activation.Conclusions：These results demonstrate that in a two-step vitrification procedure,the permeability effect in the second step is not necessary.It is possible that the second step is only required to provide adequate osmotic pressure to condense the intracellular concentration of CPAs to a level required for successful vitrification.
文摘Is there a really need to validate oocyte vitrification technique in an ART laboratory before establishing it in daily practice? Validation of micromanipulationbased technique, in this case oocyte vitrification, is essential prior to enlarging its use to routine practice. Oocyte vitrification is a new worldwide used technique and legal recently inFrance. This micromanipulation needs to be performed by a skilled and experienced embryologist and requires an internal assessment in each ART unit before any wide use. We designed a prospective study, from September 2011 to July 2012, using sibling oocytes from women who recovered more than 12 Metaphase II oocytes. A part of freshly recovered oocytes underwent immediate ICSI while the remaining oocytes were vitrified. 87 couples undergoing ICSI were selected based on number of mature oocytes available on the recovery day after denudation. A part of fresh MII oocytes were microinjected and the others were vitrified using an open system (Cryotop?). The major criterion of interest was the number of embryo transferred/ number of Metaphase II ratio for after ICSI on fresh oocytes (42/211) versus vitrified/warmed oocytes (51/204) (p > 0.05). Secondary studied criteria were survival rate (80.5% ± 26.3%), fertilization rate (68.9 ± 33.5) and finally, cumulative pregnancy rate obtained in this study is 40.2%. One of the benefits of such practice is the limitation of embryo freezing. However, the study design delays oocytes warming cycles, due to pregnancies triggered by the transfer of fresh derived oocyte embryos and to the priority to transfer all the frozen embryos before starting oocytes warming. Moreover, no data is available about children’ health. Oocyte vitrification represents not only a change in our daily practice to improve cumulative pregnancy rate but also a promising tool to develop egg banking and donation. Clinical Trials Registration number: 209 R02.
基金supported by grants from the National High Technology Research and Development Program of China(2011AA100602)the National Science and Technology Major Project of China(2013ZX08007-004,2013ZX08008-004)the National Natural Science Foundation of China(31371486)
文摘Cryopreservation techniques for mammalian oocytes and embryos have rapidly progressed during the past two decades,emphasizing their importance in various assisted reproductive technologies.Pregnancies and live births resulting from cryopreserved oocytes and embryos of several species including humans have provided proof of principle and led to the adoption of cryopreservation as an integral part of clinical in vitro fertilization.Considerable progress has been achieved in the development and application of the cryopreservation of mammalian oocytes and embryos,including preservation of the reproductive potential of patients who may become infertile,establishment of cryopreserved oocyte banks,and transport of oocytes and embryos internationally.However,the success rates are still far lower than those obtained with fresh oocytes and embryos,and there are still obstacles that need to be overcome.In this review,we address the major obstacles in the development of effective cryopreservation techniques.Such knowledge may help to eliminate these hurdles by revealing which aspects need improvement.Furthermore,this information may encourage further research by cryobiologists and increase the practical use of cryopreservation as a major part of assisted reproductive technologies for both humans and animal species.
基金performed within the framework of State projects 122041100276-0 and 075-01027-2200。
文摘Reproduction technologies(RTs)can provide for the reliable reproduction of amphibians,as well as perpetuation of species genetic variation with the use of biobanks.In 1982,in anticipation of the biodiversity conservation crisis,major Russian institutions collaborated in a dynamic program to develop and implement RTs for the sustainable management of amphibian biodiversity.An initial primary focus was the captive breeding of threatened Russian endemic anuran and caudate species,using RTs that varied from environmental manipulation to the use of exogenous gonadotropic hormones to stimulate reproduction.These species were mostly from Palearctic or cool mountain regions,but also included a wide range of species from warm regions.Other early achievements included the successful cryopreservation of anuran spermatozoa and anuran diploid pluripotent cell nuclei,in order to store both the matrilineal and patrilineal genomes in biobanks,with their subsequent development to the blastula stage after implantation into enucleated oocytes.After the turn of the 21st Century,in support of the priorities of the Amphibian Conservation Action Plan(2007),we developed RTs for the refrigerated storage of testicular or urinary spermatozoa for days to weeks at 4℃,the cryopreservation of urinary spermatozoa using anovel cryoprotectant,the in vitro fertilisation of hormonally induced oocytes either fresh or after refrigerated ex situ or in situ storage,and the artificial insemination of salamanders with fresh spermatozoa.In this article,we describe previously unpublished techniques and techniques from obscure Russian sources.
