Newcastle Fertility Center

Newcastle upon Tyne, United Kingdom

Newcastle Fertility Center

Newcastle upon Tyne, United Kingdom

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Prathalingam N.,NorthEast England Stem Cell Institute | Prathalingam N.,Northumbria University | Ferguson L.,NorthEast England Stem Cell Institute | Ferguson L.,Institute for Cellular Medicine | And 17 more authors.
Stem Cell Research and Therapy | Year: 2012

Introduction. The development of reproducible methods for deriving human embryonic stem cell (hESC) lines in compliance with good manufacturing practice (GMP) is essential for the development of hESC-based therapies. Although significant progress has been made toward the development of chemically defined conditions for the maintenance and differentiation of hESCs, efficient derivation of new hESCs requires the use of fibroblast feeder cells. However, GMP-grade feeder cell lines validated for hESC derivation are not readily available. Methods. We derived a fibroblast cell line (NclFed1A) from human foreskin in compliance with GMP standards. Consent was obtained to use the cells for the production of hESCs and to generate induced pluripotent stem cells (iPSCs). We compared the line with a variety of other cell lines for its ability to support derivation and self-renewal of hESCs. Results: NclFed1A supports efficient rates (33%) of hESC colony formation after explantation of the inner cell mass (ICM) of human blastocysts. This compared favorably with two mouse embryonic fibroblast (MEF) cell lines. NclFed1A also compared favorably with commercially available foreskin fibroblasts and MEFs in promoting proliferation and pluripotency of a number of existing and widely used hESCs. The ability of NclFed1A to maintain self-renewal remained undiminished for up to 28 population doublings from the master cell bank. Conclusions: The human fibroblast line Ncl1Fed1A, produced in compliance with GMP standards and qualified for derivation and maintenance of hESCs, is a useful resource for the advancement of progress toward hESC-based therapies in regenerative medicine. © 2012 Prathalingam et al.; licensee BioMed Central Ltd.


Panagiotopoulou N.,Newcastle upon Tyne Hospitals NHS Foundation Trust | Panagiotopoulou N.,Newcastle Fertility Center | Karavolos S.,Aberdeen Maternity Hospital | Choudhary M.,Newcastle upon Tyne Hospitals NHS Foundation Trust
European Journal of Obstetrics Gynecology and Reproductive Biology | Year: 2015

Endometrial injury to improve implantation for women undergoing assisted reproductive techniques has attracted a lot of attention recently and has rapidly become incorporated into clinical practice. The aim of this study is, thus, to assess the effectiveness and safety of endometrial injury performed in the cycle preceding assisted reproductive techniques in women with recurrent implantation failure. Electronic database searches, including MEDLINE, EMBASE, CENTRAL and grey literature, up to 30th May 2015 were conducted with no restrictions. Randomized controlled trials comparing endometrial injury versus placebo or no treatment in the cycle preceding assisted reproductive techniques in women with recurrent implantation failure were selected. The primary outcome was live birth rate. Secondary outcomes were clinical pregnancy, implantation, miscarriage and procedure-related complication rates. Of the 1115 publications identified, 4 met the inclusion criteria. Meta-analysis was not possible due to significant clinical heterogeneity among the included studies. Patients' characteristics differed, as did the intervention used with endometrial injury being performed at different phases of the preceding menstrual cycle. Moreover, the effect of endometrial injury on live birth and clinical pregnancy rates were inconsistent among the included studies. In summary, there is currently insufficient evidence to support the use of endometrial injury in women with recurrent implantation failure undergoing assisted reproductive techniques while the procedure-associated complication rate has not been assessed. Clinical implementation should, thus, be deferred until robust evidence becomes available. © 2015 Elsevier Ireland Ltd. All rights reserved. All rights reserved.


Craven L.,Institute for Ageing and Health | Tuppen H.A.,Institute for Ageing and Health | Greggains G.D.,Newcastle Fertility Center | Greggains G.D.,Institute for Ageing and Health | And 13 more authors.
Nature | Year: 2010

Mutations in mitochondrial DNA (mtDNA) are a common cause of genetic disease. Pathogenic mutations in mtDNA are detected in approximately 1 in 250 live births and at least 1 in 10,000 adults in the UK are affected by mtDNA disease. Treatment options for patients with mtDNA disease are extremely limited and are predominantly supportive in nature. Mitochondrial DNA is transmitted maternally and it has been proposed that nuclear transfer techniques may be an approach for the prevention of transmission of human mtDNA disease. Here we show that transfer of pronuclei between abnormally fertilized human zygotes results in minimal carry-over of donor zygote mtDNA and is compatible with onward development to the blastocyst stage in vitro. By optimizing the procedure we found the average level of carry-over after transfer of two pronuclei is less than 2.0%, with many of the embryos containing no detectable donor mtDNA. We believe that pronuclear transfer between zygotes, as well as the recently described metaphase II spindle transfer, has the potential to prevent the transmission of mtDNA disease in humans. © 2010 Macmillan Publishers Limited. All rights reserved.


Hyslop L.A.,Institute of Genetic Medicine | Hyslop L.A.,Newcastle Fertility Center | Blakeley P.,Francis Crick Institute | Craven L.,Northumbria University | And 26 more authors.
Nature | Year: 2016

Mitochondrial DNA (mtDNA) mutations are maternally inherited and are associated with a broad range of debilitating and fatal diseases. Reproductive technologies designed to uncouple the inheritance of mtDNA from nuclear DNA may enable affected women to have a genetically related child with a greatly reduced risk of mtDNA disease. Here we report the first preclinical studies on pronuclear transplantation (PNT). Surprisingly, techniques used in proof-of-concept studies involving abnormally fertilized human zygotes were not well tolerated by normally fertilized zygotes. We have therefore developed an alternative approach based on transplanting pronuclei shortly after completion of meiosis rather than shortly before the first mitotic division. This promotes efficient development to the blastocyst stage with no detectable effect on aneuploidy or gene expression. After optimization, mtDNA carryover was reduced to <2% in the majority (79%) of PNT blastocysts. The importance of reducing carryover to the lowest possible levels is highlighted by a progressive increase in heteroplasmy in a stem cell line derived from a PNT blastocyst with 4% mtDNA carryover. We conclude that PNT has the potential to reduce the risk of mtDNA disease, but it may not guarantee prevention. © 2016 Macmillan Publishers Limited. All rights reserved.


Lister L.M.,Northumbria University | Lister L.M.,NorthEast England Stem Cell Institute | Kouznetsova A.,Karolinska Institutet | Hyslop L.A.,Newcastle Fertility Center | And 18 more authors.
Current Biology | Year: 2010

Background: The growing trend for women to postpone childbearing has resulted in a dramatic increase in the incidence of trisomic pregnancies. Maternal age-related miscarriage and birth defects are predominantly a consequence of chromosome segregation errors during the first meiotic division (MI), which involves the segregation of replicated recombined homologous chromosomes. Despite the importance to human reproductive health, the events precipitating female age-related meiotic errors are poorly understood. Results: Here we use a long-lived wild-type mouse strain to show that the ability to segregate chromosomes synchronously during anaphase of MI declines dramatically during female aging. This is preceded by depletion of chromosome-associated cohesin in association with destabilization of chiasmata, the physical linkages between homologous chromosomes, and loss of the tight association between sister centromeres. Loss of cohesin is not due to an age-related decline in the ability of the spindle checkpoint to delay separase-mediated cleavage of cohesin until entry into anaphase I. However, we find that reduced cohesin is accompanied by depletion of Sgo2, which protects centromeric cohesin during MI. Conclusions: The data indicate that cohesin declines gradually during the long prophase arrest that precedes MI in female mammals. In aged oocytes, cohesin levels fall below the level required to stabilize chiasmata and to hold sister centromeres tightly together, leading to chromosome missegregation during MI. Cohesin loss may be amplified by a concomitant decline in the levels of the centromeric cohesin protector Sgo2. These findings indicate that cohesin is a key molecular link between female aging and chromosome missegregation during MI. © 2010 Elsevier Ltd. All rights reserved.


Hyslop L.,Newcastle Fertility Center | Prathalingam N.,Northumbria University | Nowak L.,Newcastle Fertility Center | Fenwick J.,Newcastle Fertility Center | And 8 more authors.
PLoS ONE | Year: 2012

In vitro fertilisation (IVF) and related technologies are arguably the most challenging of all cell culture applications. The starting material is a single cell from which one aims to produce an embryo capable of establishing a pregnancy eventually leading to a live birth. Laboratory processing during IVF treatment requires open manipulations of gametes and embryos, which typically involves exposure to ambient conditions. To reduce the risk of cellular stress, we have developed a totally enclosed system of interlinked isolator-based workstations designed to maintain oocytes and embryos in a physiological environment throughout the IVF process. Comparison of clinical and laboratory data before and after the introduction of the new system revealed that significantly more embryos developed to the blastocyst stage in the enclosed isolator-based system compared with conventional open-fronted laminar flow hoods. Moreover, blastocysts produced in the isolator-based system contained significantly more cells and their development was accelerated. Consistent with this, the introduction of the enclosed system was accompanied by a significant increase in the clinical pregnancy rate and in the proportion of embryos implanting following transfer to the uterus. The data indicate that protection from ambient conditions promotes improved development of human embryos. Importantly, we found that it was entirely feasible to conduct all IVF-related procedures in the isolator-based workstations. © 2012 Hyslop et al.


Herbert M.,Newcastle Fertility Center | Herbert M.,Northumbria University | Kalleas D.,Northumbria University | Cooney D.,Northumbria University | And 2 more authors.
Cold Spring Harbor Perspectives in Biology | Year: 2015

In most organisms, genome haploidization requires reciprocal DNA exchanges (crossovers) between replicated parental homologs to form bivalent chromosomes. These are resolved to their four constituent chromatids during two meiotic divisions. In female mammals, bivalents are formed during fetal life and remain intact until shortly before ovulation. Extending this period beyond ~35 years greatly increases the risk of aneuploidy in human oocytes, resulting in a dramatic increase in infertility, miscarriage, and birth defects, most notably trisomy 21. Bivalent chromosomes are stabilized by cohesion between sister chromatids, which is me-diated by the cohesin complex. In mouse oocytes, cohesin becomes depleted from chromo-somes during female aging. Consistent with this, premature loss of centromeric cohesion is a major source of aneuploidy in oocytes from older women. Here, we propose a mechanistic framework to reconcile data from genetic studies on human trisomy and oocytes with recent advances in our understanding of the molecular mechanisms of chromosome segregation during meiosis in model organisms. © 2015 Cold Spring Harbor Laboratory Press; all rights reserved.


PubMed | The Francis Crick Institute, Institute of Genetic Medicine, Oxford Business Park, Newcastle Fertility Center and 2 more.
Type: Journal Article | Journal: Nature | Year: 2016

Mitochondrial DNA (mtDNA) mutations are maternally inherited and are associated with a broad range of debilitating and fatal diseases. Reproductive technologies designed to uncouple the inheritance of mtDNA from nuclear DNA may enable affected women to have a genetically related child with a greatly reduced risk of mtDNA disease. Here we report the first preclinical studies on pronuclear transplantation (PNT). Surprisingly, techniques used in proof-of-concept studies involving abnormally fertilized human zygotes were not well tolerated by normally fertilized zygotes. We have therefore developed an alternative approach based on transplanting pronuclei shortly after completion of meiosis rather than shortly before the first mitotic division. This promotes efficient development to the blastocyst stage with no detectable effect on aneuploidy or gene expression. After optimization, mtDNA carryover was reduced to <2% in the majority (79%) of PNT blastocysts. The importance of reducing carryover to the lowest possible levels is highlighted by a progressive increase in heteroplasmy in a stem cell line derived from a PNT blastocyst with 4% mtDNA carryover. We conclude that PNT has the potential to reduce the risk of mtDNA disease, but it may not guarantee prevention.


PubMed | Newcastle Fertility Center
Type: Journal Article | Journal: PloS one | Year: 2012

In vitro fertilisation (IVF) and related technologies are arguably the most challenging of all cell culture applications. The starting material is a single cell from which one aims to produce an embryo capable of establishing a pregnancy eventually leading to a live birth. Laboratory processing during IVF treatment requires open manipulations of gametes and embryos, which typically involves exposure to ambient conditions. To reduce the risk of cellular stress, we have developed a totally enclosed system of interlinked isolator-based workstations designed to maintain oocytes and embryos in a physiological environment throughout the IVF process. Comparison of clinical and laboratory data before and after the introduction of the new system revealed that significantly more embryos developed to the blastocyst stage in the enclosed isolator-based system compared with conventional open-fronted laminar flow hoods. Moreover, blastocysts produced in the isolator-based system contained significantly more cells and their development was accelerated. Consistent with this, the introduction of the enclosed system was accompanied by a significant increase in the clinical pregnancy rate and in the proportion of embryos implanting following transfer to the uterus. The data indicate that protection from ambient conditions promotes improved development of human embryos. Importantly, we found that it was entirely feasible to conduct all IVF-related procedures in the isolator-based workstations.

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