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Devito L.,Womens Health Academic Center | Petrova A.,Womens Health Academic Center | Miere C.,Womens Health Academic Center | Codognotto S.,Womens Health Academic Center | And 6 more authors.
Stem Cells Translational Medicine | Year: 2014

Standardization guidelines for human pluripotent stem cells are still very broadly defined, despite ongoing clinical trials in the U.S., U.K., and Japan. The requirements for validation ofhumanembryonic (hESCs) and induced pluripotent stem cells (iPSCs) in general follow the regulations for other clinically compliant biologics already in place but without addressing key differences between cell types or final products. In order to realize the full potential of stem cell therapy, validation criteria, methodology, and, most importantly, strategy, should address the shortfalls and efficiency of current approaches; without this, hESC- and, especially, iPSC-based therapy will not be able to compete with other technologies in a cost-efficient way. We addressed the protocols for testing cell lines for human viral pathogens and propose a novel strategy that would significantly reduce costs. It is highly unlikely that the multiple cell lines derived in parallel from a tissue sample taken from one donor would have different profiles of endogenous viral pathogens; we therefore argue that samples from the Master Cell Banks of sibling lines could be safely pooled for validation. We illustrate this approach with tiered validation of two sibling clinical-grade hESC lines, KCL033 and KCL034 (stage 1, sterility; stage 2, specific human pathogens; and stage 3, nonspecific human pathogens). The results of all tests were negative. This cost-effective strategy could also be applied for validation of Master Cell Banks of multiple clinical-grade iPSC lines derived from a single donor. © 2014, AlphaMed Press. Source


Renwick P.,Guys And St Thomas Center For Preimplantation Genetic Diagnosis And Genetics Center | Trussler J.,Guys And St Thomas Center For Preimplantation Genetic Diagnosis And Genetics Center | Lashwood A.,Guys And St Thomas Center For Preimplantation Genetic Diagnosis And Genetics Center | Braude P.,Guys And St Thomas Center For Preimplantation Genetic Diagnosis And Genetics Center | Ogilvie C.M.,Guys And St Thomas Center For Preimplantation Genetic Diagnosis And Genetics Center
Reproductive BioMedicine Online | Year: 2010

Preimplantation genetic diagnosis using whole genome amplification and a haplotyping approach (PGH) was first described in 2006 and suggested as an efficient alternative to single-cell PCR for monogenic disorders. DNA from single cells was amplified using multiple displacement amplification; the resulting products were then tested using disease-specific PCR multiplexes applied under standard laboratory conditions to determine the haplotypes in the embryo. This study reports on a total of 127 completed biopsy cycles for 101 couples at risk of: autosomal recessive disease (71 cycles, 53 couples including one germ-line mosaic carrier), autosomal dominant disease (31 cycles, 26 couples including one germ-line mosaic carrier), X-linked recessive disease (18 cycles, 16 couples including one germ-line mosaic carrier), X-linked dominant disease (six cycles, five couples) and a double inheritance of both autosomal and X-linked recessive diseases (one cycle, one couple). Of these, 107 cycles reached embryo transfer. Overall success rates were: fetal heart beat-positive pregnancies (FHB+)/biopsy cycle = 28%; FHB+/embryo transfer = 34%; FHB+/couple = 36%; 26 babies born, 13 ongoing pregnancies. These data demonstrate that PGH provides a robust, efficient and successful alternative to single-cell PCR for monogenic diseases. © 2010, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved. Source

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