Derivation of embryonic stem cells from pre-implantation genetic diagnosis embryos: possible but inefficient

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OBJECTIVE: Disease specific embryonic stem cells (ESC) have been derived from affected embryos undergoing pre-implantation genetic diagnosis (PGD). However, the likelihood of creating an ESC line from a potential donor is unknown. Factors throughout the process limit the efficiency of ESC generation. Techniques to optimize ESC derivation from non-PGD embryos have been established, though whether they apply to PGD embryos is unclear. This information would be useful to patients considering donation as well as their clinicians. We report on our experience deriving ESC from our PGD program. DESIGN: Descriptive study. MATERIALS AND METHODS: Patients undergoing PGD were recruited to donate affected embryos. Embryos were graded and cultured until day 6. The inner cell mass (ICM) was isolated by laser and plated on feeder cells. Cultures were assessed regularly for adherence and growth. hESC were characterized using standard techniques. RESULTS: From 9/08 to 6/09, we initiated 140 PGD cycles: 39 for single gene disorders, 22 for translocations, and 79 for aneuploidy screening. 12 patients consented to participate and 4 ultimately donated. Consented subjects who did not donate had cancelled cycles or no viable embryos. 36 embryos were obtained ranging in grade from cleavage stage arrest to 6AA blastocysts. 30 ICMs were plated, of which 12 adhered. 3 ESC lines were derived, 2 from a patient with Charcot Marie Tooth and 1 from a patient with a translocation. CONCLUSION: ESC lines can be derived from PGD embryos, though with limited efficiency. In this study, 3 ESC lines were derived from a large potential group of donors. While our embryo-specific derivation rates are lower than those reported for non-PGD embryos, whether this is operator or embryo dependent is unclear. ESC yield from PGD cases may be limited by other factors such as, PGD indication, willingness to donate to research, and cycle outcome. Further study is needed to clarify these issues and optimize the creation of ESC lines.