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Sally Catt

Monash University, Obstetrics and Gynaecology, Faculty Member

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Address: Cheltenham, Victoria, Australia

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Papers by Sally Catt

Effects of COVID-19 Quarantine Period on Fertility Patients and IVF Clinic Management Worldwide

Fertility & reproduction, Sep 1, 2022

Non-Invasive Metabolic Live Cell Imaging of Early Embryo Development Using Novel Adapted Confocal Microscopy in Ageing

Fertility & Reproduction

Background: Developing non-invasive methods that are reliable to assess embryo quality has been a... more Background: Developing non-invasive methods that are reliable to assess embryo quality has been a significant aim for assisted reproductive technologies. Changes in metabolic activity could lead to abnormal embryo development and implantation potential which could be potentially predicted by incorporating non-invasive measurements of metabolism. Metabolic imaging of nicotinamide adenine dinucleotide (NADH) has been well studied in cell lines, however, evidence for its use in embryo development is limited. Measurements of NADH during advanced-reproductive-age could be a useful tool as the auto-fluorescence of NADH is a straightforward representation of mitochondrial function and cellular redox status. Aim: To determine potential differences in NADH levels associated in early embryo development from young and old mice using novel adapted confocal microscopy. Methods: Oocytes from two groups of 20 super-ovulated female mice (Young: 5-8 weeks old, n=10; Old: 45-52 weeks old, n=10) were ...

Warming Oocytes with a Universal Warming Medium and Protocol is as Good as the Company Media and Protocols

Fertility & Reproduction

Background: A universal warming media and protocol would simplify exchange and use of oocytes and... more Background: A universal warming media and protocol would simplify exchange and use of oocytes and embryos between different IVF centres. Aim: The aim of this study was to compare a universal warming medium and protocol (Parmegiani et al, 2014 RBMonline 28, 614-23) with 3 other warming media and protocols which were used with their corresponding vitrification media (2 commercial, one in house) measuring survival (pre and post insemination combined), fertilization and blastocyst rate. Method: This was a prospective study on 300 mature cumulus-free F1 mouse oocytes divided between three vitrification protocols, SAGE™ vitrification (S), RapidVit™ Omni by Vitrolife (V), and the in house ‘MCE-Trehalose’ (M) designed for mouse embryos. Oocytes for each vitrification group (S,V & M) were divided equally to be warmed either by the corresponding warming protocols (S/S, V/V & M/M and or the universal warming protocol (S/U, V/U and M/U). Surviving oocytes were inseminated in vitro with 250,000 ...

DNA repair and response to sperm DNA damage in oocytes and embryos, and the potential consequences in ART: a systematic review

Molecular Human Reproduction, 2021

Sperm DNA damage is considered a predictive factor for the clinical outcomes of patients undergoi... more Sperm DNA damage is considered a predictive factor for the clinical outcomes of patients undergoing ART. Laboratory evidence suggests that zygotes and developing embryos have adopted specific response and repair mechanisms to repair DNA damage of paternal origin. We have conducted a systematic review in accordance with guidelines from Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) to identify and review the maternal mechanisms used to respond and repair sperm DNA damage during early embryonic development, how these mechanisms operate and their potential clinical implications. The literature search was conducted in Ovid MEDLINE and Embase databases until May 2021. Out of 6297 articles initially identified, 36 studies were found to be relevant through cross referencing and were fully extracted. The collective evidence in human and animal models indicate that the early embryo has the capacity to repair DNA damage within sperm by activating maternally driven...

P–790 Effects of COVID–19 quarantine period on Fertility Treatment and IVF Clinic management

Human Reproduction, 2021

Study question What are the effects of the initial COVID–19 response on the management of fertili... more Study question What are the effects of the initial COVID–19 response on the management of fertility clinics and clinical practice around the world? Summary answer In the COVID–19 outbreak, the large majority of fertility clinics worldwide suspended fertility treatments. In cycles that continued, there was a shift to frozen embryo-transfer. What is known already After the initial months of 2020 showed a rapid spread of the new Coronavirus SARS-CoV–2, the World Health Organisation declared a global pandemic on 11 March 2020. Occupation of health care facilities with acutely sick patients and the need to reduce infection transmission led to a reduction in capacity to perform elective medical procedures. Little was known on the global impact of COVID–19 on fertility care. With the implication of ‘lockdowns’ in different countries around the world to stop the spread of the virus, the question was posed on how fertility clinics and treatments would proceed moving forward. Study design, si...

An in vitro assessment of the value of sucrose supplementation to warming media for the vitrification of bovine embryos

Cryo letters, 2017

　　BACKGROUND: In order to thaw slow-cooled bovine embryos it is standard practice to draw out per... more 　　BACKGROUND: In order to thaw slow-cooled bovine embryos it is standard practice to draw out permeating cryoprotectants by passing embryos through successively decreasing osmotic solutions. However, recently it has been suggested that sucrose may not be needed in the warming media. OBJECTIVE The aim of this experiment was to compare the effect of warming media prepared with or without the inclusion of sucrose on the survival and hatching capacity of vitrified in vitro-derived bovine embryos. MATERIALS AND METHODS Expanded blastocysts were produced in vitro and vitrified. Vitrified embryos were warmed either successively through 0.5, 0.3 and 0.2 M sucrose solutions ('stepwise'), or by placing directly into the blastocyst solution without the addition of sucrose ('direct'). A total of 93 expanded blastocysts were assigned randomly to two treatment groups, respectively. RESULTS The re-expansion rates of vitrified embryos warmed after 24h in vitro culture were similar b...

Vitrification of in vitro-derived bovine embryos: targeting enhancement of quality by refining technology and standardising procedures

Reproduction, Fertility and Development, 2019

Bovine invitro fertilisation technology has been widely exploited in commercial settings. The maj... more Bovine invitro fertilisation technology has been widely exploited in commercial settings. The majority of invitro-derived cattle embryos are transferred into recipient cows as recently collected (i.e. ‘fresh’) embryos due to the lack of a reliable cryopreservation method that results in favourable pregnancy rates following transfer of thawed embryos. This is a primary reason for the poor industry uptake of this extreme temperature freezing process. Numerous investigations into vitrification have revealed the importance of rapid cooling and warming rates, enhancing embryo viability after cryopreservation compared with conventional slow freezing. Those studies spawned a considerable assortment of cryovessels and diversity of procedures, delivering variable rates of success, which makes performing vitrification consistently a practical challenge. Hence, further research is required in order to both optimise and standardise vitrification methodology and to design a cryovessel that enabl...

Podocalyxin molecular characteristics and endometrial expression: high conservation between humans and macaques but divergence in mice

Biology of Reproduction, 2022

Podocalyxin (PODXL) is a newly identified key negative regulator of human endometrial receptivity... more Podocalyxin (PODXL) is a newly identified key negative regulator of human endometrial receptivity, specifically down-regulated in the luminal epithelium at receptivity to permit embryo implantation. Here, we bioinformatically compared the molecular characteristics of PODXL among the human, rhesus macaque and mouse, determined by immunohistochemistry and in situ hybridization (mouse tissues) whether endometrial PODXL expression is conserved across the three species, and examined if PODXL inhibits mouse embryo attachment in vitro. The PODXL gene, mRNA and protein sequences showed greater similarities between humans and macaques than with mice. In all species, PODXL was expressed in endometrial luminal/glandular epithelia and endothelia. In macaques (n = 9), luminal PODXL was significantly down-regulated when receptivity is developed, consistent with the pattern found in women. At receptivity PODXL was also reduced in shallow glands, whereas endothelial expression was unchanged across ...

The Effects of FSH and Glutathione Supplementation to the in Vitro Maturation Media on Mouse Oocyte Maturation

Proceedings of the Proceedings of the 1st EAI International Conference on Medical And Health Research, ICoMHER November 13-14th 2018, Padang, West Sumatera, Indonesia, 2019

Either Follicle-stimulating hormone (FSH) or glutathione solely has been known to have benefits o... more Either Follicle-stimulating hormone (FSH) or glutathione solely has been known to have benefits on oocyte maturation, yet study about combination effects are still limited. This study aimed to observe the effects of FSH and glutathione supplementation to the in vitro maturation (IVM) media on mouse oocyte maturation. Immature oocytes of F1 (C57BL/6xCBA) female mice were matured for 24 hours in Minimum Essential Medium (MEM) Alpha supplemented with 5 μg/mL FSH and 0.15 mM glutathione, in 3 different combinations, i.e. the MEM alpha without any supplementations, supplementation of FSH solely, and combination of FSH with glutathione. A total of 209 immature oocytes were studied to examine the oocyte maturation. Treatment with FSH supplementation alone (p=0.043) and in combination with glutathione (p=0.016) showed significantly higher oocyte maturation. This study indicated that in vitro oocyte maturation can be augmented by supplementation of IVM medium with FSH, either solely or along with glutathione.

A comparative analysis of the efficacy of three cryopreservation protocols on the survival of in vitro-derived cattle embryos at pronuclear and blastocyst stages

Cryobiology, Aug 1, 2017

The effectiveness of three cryopreservation protocols (slow freezing, short equilibration vitrifi... more The effectiveness of three cryopreservation protocols (slow freezing, short equilibration vitrification and long equilibration vitrification) on in vitro-derived cattle embryos at expanded blastocyst and pronuclear stages was compared. 199 expanded blastocysts of good quality were assigned randomly into four treatment groups [control, non-cryopreserved (fresh, unfrozen); and the three cryopreservation methods]. The re-expansion of the cryopreserved blastocysts after 24 h in vitro culture was similar to that of the fresh control group. However, the hatching rate of expanded blastocysts after 48 h culture was significantly less for the slow freezing group (31/47; 66.0%) than for both the short equilibration vitrification (46/51; 90.2%) and long equilibration vitrification groups (42/50; 84.0%). Denuded presumptive zygotes at the pronuclear stage (14-18 h post-insemination) were assigned randomly to the same four treatment groups and, following thawing, embryos were assessed for their ...

Effect of Chemically Defined Lipid Molecules on Blastocyst Development and Outgrowth

International Journal of Anatomy and Research, 2016

Fatty acids play a diverse role in early embryonic development and implantation. This study exami... more Fatty acids play a diverse role in early embryonic development and implantation. This study examined the effect of a chemically defined additive containing saturated and unsaturated fatty acids on mouse fertilization and embryo development in vitro. Objective: To observe the effect of a chemically defined addition of fatty acids on in-vitro fertilization, early embryonic development and blastocyst outgrowth Study Design: F1 mice (4-6 weeks) were superovulated and 16 hours later oocytes were collected and prepared for IVF using spermatozoa from F1 males. Oocytes (n=355) were randomly allocated to global fertilization medium with (+) or without (-) chemically defined lipid (#11905-031, Life Global Technologies). Fertilization was checked (2-PN) 6-9hrs after insemination and embryos were grown up in blastocyst culture medium + lipid additive or controls. Embryo development was monitored daily until D5 when blastocysts were transferred to 96well plates. Blastocyst outgrowths were imaged on D6, D7 and D9 using XY-Cyclone software. Data was analyzed (Chi-squares) to compare the effect of lipid on fertilization rate, blastocyst development and outgrowths. Result: Oocytes (n=355) randomly allocated to control (n=170) or +lipid groups for fertilization showed no difference in fertilization rates in medium with (n=155) or without lipid. Addition of lipid to blastocyst medium resulted in a significant increase (p<0.05) in blastocyst formation (73% vs 87%). but produced no difference in blastocyst hatching or outgrowth area. Conclusion: A chemically defined lipid additive promoted embryonic development in vitro. Further studies are needed to examine the dose-response effect of lipid on fertilization and embryo development.

The impact of COVID-19 mitigation measures on fertility patients and clinics around the world

Reproductive BioMedicine Online

A human-based assisted reproduction protocol for the menstruating spiny mouse, Acomys cahirinus

PLoS ONE, 2020

The Egyptian or Common spiny mouse (A. cahirinus) is the first rodent species to show human-like ... more The Egyptian or Common spiny mouse (A. cahirinus) is the first rodent species to show human-like menstruation and spontaneous decidualisation. We consider from these, and its other, human-like characteristics that this species will be a more useful and appropriate small animal model for human reproductive studies. Based on this, there is a need to develop specific laboratory-based assisted reproduction protocols including superovulation, in-vitro fertilisation, embryo cryopreservation and transfer to expand and make this model more relevant. Because standard rodent superovulation has not been successful in the spiny mouse, we have selected to test a human protocol. Female spiny mice will receive a subcutaneous GnRH agonist implant and be allowed to recover. Menstrual cycle lengths will then be allowed to stabilize prior to ovarian stimulation. After recovery, females will be injected IP once a day for 4 days with a FSH analogue, to induce follicular growth, and on day 5 will be inje...

A randomised trial investigating the effectiveness and safety of three IVF laboratory disinfectants

Human Reproduction, 2013

The rate of development of in vitro -derived cattle embryos affects neither their survival nor their hatching rate following recovery from cryopreservation by vitrification

F1000Research, 2018

Cyclic nucleotide in oocyte In vitro maturation in Assisted Reproductive Technology

Article Info History Received: 07 Dec 2020 Accepted: 23 Dec 2020 Available: 31 Dec 2020 Abstract ... more Article Info History Received: 07 Dec 2020 Accepted: 23 Dec 2020 Available: 31 Dec 2020 Abstract In vitro maturation (IVM) is a promising assisted reproductive technology (ART) for human infertility treatment. However, when cumulus oocyte complexes (COCs) are removed from their follicular environment when manipulated in vitro, it can lead to a decrease of intra-oocyte cyclic adenosine 3’, 5’-monophosphare (cAMP) causing spontaneous nuclear maturation and an asynchrony with the oocytes’ cytoplasmic maturation, resulting in poor embryo developmental outcomes. Nuclear and cytoplasmic synchrony is important during oocyte maturation within antral follicles. It is maintained partially by the actions of c-type natriuretic peptide (CNP) binding with natriuretic peptide receptor 2 (NPR2), supporting high cAMP levels thus holding the oocyte in meiotic arrest. Addition of CNP to pre-IVM media has the capacity of maintaining cAMP levels and thus improve synchrony. Moreover, in women with advanc...

Male ageing negatively affects the chances of live birth in IVF/ICSI cycles for idiopathic infertility

Cryopreserved Ovarian Tissue Grafts of Adult Mice

Toxic town: ART disinfectants damage human sperms: An investigation into potential toxicity of ART disinfectants on human sperms

Aim: To investigate toxicity of disinfectants using human sperm motility assay (HSMA) in order to... more Aim: To investigate toxicity of disinfectants using human sperm motility assay (HSMA) in order to establish universal disinfection protocol, safe by all means for ART set up. Methods: Semen samples obtained from normospermic men and processed through density gradient, pellet was resuspended in 5ml of KSOM handling medium and motility assessed at 0 hours. The replicate splitted equally into five 1ml aliquots.In direct contact trial, four test tubes treated by adding 0.5ml of each disinfectant simultaneously ( 70% Ethanol, Fertisafe, Fertisafe Plus & Farmacidal ) directly, one test tube without disinfectant being a control. Results: Human sperm motility was lost after 24hrs of direct contact trial treatment with disinfectants as compared to control (p<0.01). Sperm motility lost was more than 90 percent after 24hrs of direct contacttrial of disinfectants. Conclusion: Significant association of disinfectant use in ART setup and loss of sperm motility was found in direct contacttrial.

Requirements for Cryopreservation of In vitro-Produced Bovine Embryos by a Standard Method of Vitrification

Journal of Veterinary Science and Animal Husbandry

In relation to assisted mammalian reproduction, the goal of cryopreservation is to preserve witho... more In relation to assisted mammalian reproduction, the goal of cryopreservation is to preserve without significant loss of viability a stock of gametes and/or embryos with a view to thawing those cells for use in in vitro reproduction treatments. There are numerous cryopreservation protocols, which vary in terms of cryoprotectant used, storage temperature, freezing and thawing rates, and the particular cells that they are suitable for preserving. Although slow freezing has become a standard method for in vivo bovine embryo cryopreservation, it seems not to be efficient for preserving in vitro-produced (IVP) bovine embryos. Over the past decade, vitrification, a process of glass-like solidification, has become the cryopreservation method of choice for human oocytes and embryos. This is because it is often less time-consuming than slow freezing, does not require expensive 'slow rate' freezing machines, and has been proven to increase survival rates compared to slow frozen embryos. In contrast to clinical applications, in the cattle industry vitrification presents shortcomings, especially when applied to IVP embryos. A high concentration of cryoprotectant in solution is needed to induce vitrification. However, if left too long with metabolizing embryos, cryoprotectants can become toxic. Failure to standardize vitrification protocols leads to inconsistent results between laboratories, making application less practical in field settings. Therefore, determination of the most suitable vitrification method is important to advance its routine commercial use. Moreover, simplification of the vitrification procedure through development of an in-straw dilution without the use of a microscope may help to expand the use of vitrification methods on the farm.

Effects of COVID-19 Quarantine Period on Fertility Patients and IVF Clinic Management Worldwide

Fertility & reproduction, Sep 1, 2022

Non-Invasive Metabolic Live Cell Imaging of Early Embryo Development Using Novel Adapted Confocal Microscopy in Ageing

Fertility & Reproduction

Background: Developing non-invasive methods that are reliable to assess embryo quality has been a... more Background: Developing non-invasive methods that are reliable to assess embryo quality has been a significant aim for assisted reproductive technologies. Changes in metabolic activity could lead to abnormal embryo development and implantation potential which could be potentially predicted by incorporating non-invasive measurements of metabolism. Metabolic imaging of nicotinamide adenine dinucleotide (NADH) has been well studied in cell lines, however, evidence for its use in embryo development is limited. Measurements of NADH during advanced-reproductive-age could be a useful tool as the auto-fluorescence of NADH is a straightforward representation of mitochondrial function and cellular redox status. Aim: To determine potential differences in NADH levels associated in early embryo development from young and old mice using novel adapted confocal microscopy. Methods: Oocytes from two groups of 20 super-ovulated female mice (Young: 5-8 weeks old, n=10; Old: 45-52 weeks old, n=10) were ...

Warming Oocytes with a Universal Warming Medium and Protocol is as Good as the Company Media and Protocols

Fertility & Reproduction

Background: A universal warming media and protocol would simplify exchange and use of oocytes and... more Background: A universal warming media and protocol would simplify exchange and use of oocytes and embryos between different IVF centres. Aim: The aim of this study was to compare a universal warming medium and protocol (Parmegiani et al, 2014 RBMonline 28, 614-23) with 3 other warming media and protocols which were used with their corresponding vitrification media (2 commercial, one in house) measuring survival (pre and post insemination combined), fertilization and blastocyst rate. Method: This was a prospective study on 300 mature cumulus-free F1 mouse oocytes divided between three vitrification protocols, SAGE™ vitrification (S), RapidVit™ Omni by Vitrolife (V), and the in house ‘MCE-Trehalose’ (M) designed for mouse embryos. Oocytes for each vitrification group (S,V & M) were divided equally to be warmed either by the corresponding warming protocols (S/S, V/V & M/M and or the universal warming protocol (S/U, V/U and M/U). Surviving oocytes were inseminated in vitro with 250,000 ...

DNA repair and response to sperm DNA damage in oocytes and embryos, and the potential consequences in ART: a systematic review

Molecular Human Reproduction, 2021

Sperm DNA damage is considered a predictive factor for the clinical outcomes of patients undergoi... more Sperm DNA damage is considered a predictive factor for the clinical outcomes of patients undergoing ART. Laboratory evidence suggests that zygotes and developing embryos have adopted specific response and repair mechanisms to repair DNA damage of paternal origin. We have conducted a systematic review in accordance with guidelines from Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) to identify and review the maternal mechanisms used to respond and repair sperm DNA damage during early embryonic development, how these mechanisms operate and their potential clinical implications. The literature search was conducted in Ovid MEDLINE and Embase databases until May 2021. Out of 6297 articles initially identified, 36 studies were found to be relevant through cross referencing and were fully extracted. The collective evidence in human and animal models indicate that the early embryo has the capacity to repair DNA damage within sperm by activating maternally driven...

P–790 Effects of COVID–19 quarantine period on Fertility Treatment and IVF Clinic management

Human Reproduction, 2021

Study question What are the effects of the initial COVID–19 response on the management of fertili... more Study question What are the effects of the initial COVID–19 response on the management of fertility clinics and clinical practice around the world? Summary answer In the COVID–19 outbreak, the large majority of fertility clinics worldwide suspended fertility treatments. In cycles that continued, there was a shift to frozen embryo-transfer. What is known already After the initial months of 2020 showed a rapid spread of the new Coronavirus SARS-CoV–2, the World Health Organisation declared a global pandemic on 11 March 2020. Occupation of health care facilities with acutely sick patients and the need to reduce infection transmission led to a reduction in capacity to perform elective medical procedures. Little was known on the global impact of COVID–19 on fertility care. With the implication of ‘lockdowns’ in different countries around the world to stop the spread of the virus, the question was posed on how fertility clinics and treatments would proceed moving forward. Study design, si...

An in vitro assessment of the value of sucrose supplementation to warming media for the vitrification of bovine embryos

Cryo letters, 2017

　　BACKGROUND: In order to thaw slow-cooled bovine embryos it is standard practice to draw out per... more 　　BACKGROUND: In order to thaw slow-cooled bovine embryos it is standard practice to draw out permeating cryoprotectants by passing embryos through successively decreasing osmotic solutions. However, recently it has been suggested that sucrose may not be needed in the warming media. OBJECTIVE The aim of this experiment was to compare the effect of warming media prepared with or without the inclusion of sucrose on the survival and hatching capacity of vitrified in vitro-derived bovine embryos. MATERIALS AND METHODS Expanded blastocysts were produced in vitro and vitrified. Vitrified embryos were warmed either successively through 0.5, 0.3 and 0.2 M sucrose solutions ('stepwise'), or by placing directly into the blastocyst solution without the addition of sucrose ('direct'). A total of 93 expanded blastocysts were assigned randomly to two treatment groups, respectively. RESULTS The re-expansion rates of vitrified embryos warmed after 24h in vitro culture were similar b...

Vitrification of in vitro-derived bovine embryos: targeting enhancement of quality by refining technology and standardising procedures

Reproduction, Fertility and Development, 2019

Bovine invitro fertilisation technology has been widely exploited in commercial settings. The maj... more Bovine invitro fertilisation technology has been widely exploited in commercial settings. The majority of invitro-derived cattle embryos are transferred into recipient cows as recently collected (i.e. ‘fresh’) embryos due to the lack of a reliable cryopreservation method that results in favourable pregnancy rates following transfer of thawed embryos. This is a primary reason for the poor industry uptake of this extreme temperature freezing process. Numerous investigations into vitrification have revealed the importance of rapid cooling and warming rates, enhancing embryo viability after cryopreservation compared with conventional slow freezing. Those studies spawned a considerable assortment of cryovessels and diversity of procedures, delivering variable rates of success, which makes performing vitrification consistently a practical challenge. Hence, further research is required in order to both optimise and standardise vitrification methodology and to design a cryovessel that enabl...

Podocalyxin molecular characteristics and endometrial expression: high conservation between humans and macaques but divergence in mice

Biology of Reproduction, 2022

Podocalyxin (PODXL) is a newly identified key negative regulator of human endometrial receptivity... more Podocalyxin (PODXL) is a newly identified key negative regulator of human endometrial receptivity, specifically down-regulated in the luminal epithelium at receptivity to permit embryo implantation. Here, we bioinformatically compared the molecular characteristics of PODXL among the human, rhesus macaque and mouse, determined by immunohistochemistry and in situ hybridization (mouse tissues) whether endometrial PODXL expression is conserved across the three species, and examined if PODXL inhibits mouse embryo attachment in vitro. The PODXL gene, mRNA and protein sequences showed greater similarities between humans and macaques than with mice. In all species, PODXL was expressed in endometrial luminal/glandular epithelia and endothelia. In macaques (n = 9), luminal PODXL was significantly down-regulated when receptivity is developed, consistent with the pattern found in women. At receptivity PODXL was also reduced in shallow glands, whereas endothelial expression was unchanged across ...

The Effects of FSH and Glutathione Supplementation to the in Vitro Maturation Media on Mouse Oocyte Maturation

Proceedings of the Proceedings of the 1st EAI International Conference on Medical And Health Research, ICoMHER November 13-14th 2018, Padang, West Sumatera, Indonesia, 2019

Either Follicle-stimulating hormone (FSH) or glutathione solely has been known to have benefits o... more Either Follicle-stimulating hormone (FSH) or glutathione solely has been known to have benefits on oocyte maturation, yet study about combination effects are still limited. This study aimed to observe the effects of FSH and glutathione supplementation to the in vitro maturation (IVM) media on mouse oocyte maturation. Immature oocytes of F1 (C57BL/6xCBA) female mice were matured for 24 hours in Minimum Essential Medium (MEM) Alpha supplemented with 5 μg/mL FSH and 0.15 mM glutathione, in 3 different combinations, i.e. the MEM alpha without any supplementations, supplementation of FSH solely, and combination of FSH with glutathione. A total of 209 immature oocytes were studied to examine the oocyte maturation. Treatment with FSH supplementation alone (p=0.043) and in combination with glutathione (p=0.016) showed significantly higher oocyte maturation. This study indicated that in vitro oocyte maturation can be augmented by supplementation of IVM medium with FSH, either solely or along with glutathione.

A comparative analysis of the efficacy of three cryopreservation protocols on the survival of in vitro-derived cattle embryos at pronuclear and blastocyst stages

Cryobiology, Aug 1, 2017

The effectiveness of three cryopreservation protocols (slow freezing, short equilibration vitrifi... more The effectiveness of three cryopreservation protocols (slow freezing, short equilibration vitrification and long equilibration vitrification) on in vitro-derived cattle embryos at expanded blastocyst and pronuclear stages was compared. 199 expanded blastocysts of good quality were assigned randomly into four treatment groups [control, non-cryopreserved (fresh, unfrozen); and the three cryopreservation methods]. The re-expansion of the cryopreserved blastocysts after 24 h in vitro culture was similar to that of the fresh control group. However, the hatching rate of expanded blastocysts after 48 h culture was significantly less for the slow freezing group (31/47; 66.0%) than for both the short equilibration vitrification (46/51; 90.2%) and long equilibration vitrification groups (42/50; 84.0%). Denuded presumptive zygotes at the pronuclear stage (14-18 h post-insemination) were assigned randomly to the same four treatment groups and, following thawing, embryos were assessed for their ...

Effect of Chemically Defined Lipid Molecules on Blastocyst Development and Outgrowth

International Journal of Anatomy and Research, 2016

Fatty acids play a diverse role in early embryonic development and implantation. This study exami... more Fatty acids play a diverse role in early embryonic development and implantation. This study examined the effect of a chemically defined additive containing saturated and unsaturated fatty acids on mouse fertilization and embryo development in vitro. Objective: To observe the effect of a chemically defined addition of fatty acids on in-vitro fertilization, early embryonic development and blastocyst outgrowth Study Design: F1 mice (4-6 weeks) were superovulated and 16 hours later oocytes were collected and prepared for IVF using spermatozoa from F1 males. Oocytes (n=355) were randomly allocated to global fertilization medium with (+) or without (-) chemically defined lipid (#11905-031, Life Global Technologies). Fertilization was checked (2-PN) 6-9hrs after insemination and embryos were grown up in blastocyst culture medium + lipid additive or controls. Embryo development was monitored daily until D5 when blastocysts were transferred to 96well plates. Blastocyst outgrowths were imaged on D6, D7 and D9 using XY-Cyclone software. Data was analyzed (Chi-squares) to compare the effect of lipid on fertilization rate, blastocyst development and outgrowths. Result: Oocytes (n=355) randomly allocated to control (n=170) or +lipid groups for fertilization showed no difference in fertilization rates in medium with (n=155) or without lipid. Addition of lipid to blastocyst medium resulted in a significant increase (p<0.05) in blastocyst formation (73% vs 87%). but produced no difference in blastocyst hatching or outgrowth area. Conclusion: A chemically defined lipid additive promoted embryonic development in vitro. Further studies are needed to examine the dose-response effect of lipid on fertilization and embryo development.

The impact of COVID-19 mitigation measures on fertility patients and clinics around the world

Reproductive BioMedicine Online

A human-based assisted reproduction protocol for the menstruating spiny mouse, Acomys cahirinus

PLoS ONE, 2020

The Egyptian or Common spiny mouse (A. cahirinus) is the first rodent species to show human-like ... more The Egyptian or Common spiny mouse (A. cahirinus) is the first rodent species to show human-like menstruation and spontaneous decidualisation. We consider from these, and its other, human-like characteristics that this species will be a more useful and appropriate small animal model for human reproductive studies. Based on this, there is a need to develop specific laboratory-based assisted reproduction protocols including superovulation, in-vitro fertilisation, embryo cryopreservation and transfer to expand and make this model more relevant. Because standard rodent superovulation has not been successful in the spiny mouse, we have selected to test a human protocol. Female spiny mice will receive a subcutaneous GnRH agonist implant and be allowed to recover. Menstrual cycle lengths will then be allowed to stabilize prior to ovarian stimulation. After recovery, females will be injected IP once a day for 4 days with a FSH analogue, to induce follicular growth, and on day 5 will be inje...

A randomised trial investigating the effectiveness and safety of three IVF laboratory disinfectants

Human Reproduction, 2013

The rate of development of in vitro -derived cattle embryos affects neither their survival nor their hatching rate following recovery from cryopreservation by vitrification

F1000Research, 2018

Cyclic nucleotide in oocyte In vitro maturation in Assisted Reproductive Technology

Article Info History Received: 07 Dec 2020 Accepted: 23 Dec 2020 Available: 31 Dec 2020 Abstract ... more Article Info History Received: 07 Dec 2020 Accepted: 23 Dec 2020 Available: 31 Dec 2020 Abstract In vitro maturation (IVM) is a promising assisted reproductive technology (ART) for human infertility treatment. However, when cumulus oocyte complexes (COCs) are removed from their follicular environment when manipulated in vitro, it can lead to a decrease of intra-oocyte cyclic adenosine 3’, 5’-monophosphare (cAMP) causing spontaneous nuclear maturation and an asynchrony with the oocytes’ cytoplasmic maturation, resulting in poor embryo developmental outcomes. Nuclear and cytoplasmic synchrony is important during oocyte maturation within antral follicles. It is maintained partially by the actions of c-type natriuretic peptide (CNP) binding with natriuretic peptide receptor 2 (NPR2), supporting high cAMP levels thus holding the oocyte in meiotic arrest. Addition of CNP to pre-IVM media has the capacity of maintaining cAMP levels and thus improve synchrony. Moreover, in women with advanc...

Male ageing negatively affects the chances of live birth in IVF/ICSI cycles for idiopathic infertility

Cryopreserved Ovarian Tissue Grafts of Adult Mice

Toxic town: ART disinfectants damage human sperms: An investigation into potential toxicity of ART disinfectants on human sperms

Aim: To investigate toxicity of disinfectants using human sperm motility assay (HSMA) in order to... more Aim: To investigate toxicity of disinfectants using human sperm motility assay (HSMA) in order to establish universal disinfection protocol, safe by all means for ART set up. Methods: Semen samples obtained from normospermic men and processed through density gradient, pellet was resuspended in 5ml of KSOM handling medium and motility assessed at 0 hours. The replicate splitted equally into five 1ml aliquots.In direct contact trial, four test tubes treated by adding 0.5ml of each disinfectant simultaneously ( 70% Ethanol, Fertisafe, Fertisafe Plus & Farmacidal ) directly, one test tube without disinfectant being a control. Results: Human sperm motility was lost after 24hrs of direct contact trial treatment with disinfectants as compared to control (p<0.01). Sperm motility lost was more than 90 percent after 24hrs of direct contacttrial of disinfectants. Conclusion: Significant association of disinfectant use in ART setup and loss of sperm motility was found in direct contacttrial.

Requirements for Cryopreservation of In vitro-Produced Bovine Embryos by a Standard Method of Vitrification

Journal of Veterinary Science and Animal Husbandry

In relation to assisted mammalian reproduction, the goal of cryopreservation is to preserve witho... more In relation to assisted mammalian reproduction, the goal of cryopreservation is to preserve without significant loss of viability a stock of gametes and/or embryos with a view to thawing those cells for use in in vitro reproduction treatments. There are numerous cryopreservation protocols, which vary in terms of cryoprotectant used, storage temperature, freezing and thawing rates, and the particular cells that they are suitable for preserving. Although slow freezing has become a standard method for in vivo bovine embryo cryopreservation, it seems not to be efficient for preserving in vitro-produced (IVP) bovine embryos. Over the past decade, vitrification, a process of glass-like solidification, has become the cryopreservation method of choice for human oocytes and embryos. This is because it is often less time-consuming than slow freezing, does not require expensive 'slow rate' freezing machines, and has been proven to increase survival rates compared to slow frozen embryos. In contrast to clinical applications, in the cattle industry vitrification presents shortcomings, especially when applied to IVP embryos. A high concentration of cryoprotectant in solution is needed to induce vitrification. However, if left too long with metabolizing embryos, cryoprotectants can become toxic. Failure to standardize vitrification protocols leads to inconsistent results between laboratories, making application less practical in field settings. Therefore, determination of the most suitable vitrification method is important to advance its routine commercial use. Moreover, simplification of the vitrification procedure through development of an in-straw dilution without the use of a microscope may help to expand the use of vitrification methods on the farm.