Neonatal outcomes of live births after blastocyst biopsy in preimplantation genetic testing cycles: a follow-up of 1,721 children.

OBJECTIVE: To investigate whether blastocyst biopsy in preimplantation genetic testing (PGT) increases the risk of adverse neonatal outcomes. DESIGN: Retrospective cohort study. SETTING: University-affiliated center. PATIENTS: Live births after blastocyst biopsy combined with frozen ET (PGT group) and frozen blastocyst transfer after in vitro fertilization or intracytoplasmic sperm injection (control group). INTERVENTION(S): Blastocyst biopsy. MAIN OUTCOME MEASURE(S): Gestational age (GA), birth weight (BW), and rates of preterm birth (PB), very preterm birth (VPB), extreme preterm birth (EPB), low birth weight (LBW), very low birth weight (VLBW), and macrosomia. RESULT(S): No significant differences were observed in the sex ratio, GA, PB, VPB, EPB, BW, or rates of LBW, VLBW, and macrosomia between the PGT and control groups for either singletons or twins. However, the cesarean section rate of the PGT group was significantly higher than that of the control group for twins (adjusted odds ratio, 2.383 [1.079, 5.259]). Regarding fluorescence in situ hybridization-PGT neonates, neonatal outcomes, including GA, BW, and rates of PB, VPB, LBW, and VLBW, did not differ between the different groups of biopsied cells (≥10 group and <10 group) for either the grade B or grade C trophectoderm score subgroups; however, in the grade B trophectoderm score subgroup, the rate of boy babies in the ≥10 group was significantly higher than that in the <10 group (83.3% vs. 40.9%). The association between the number of biopsied cells and GA/BW was not statistically significant. CONCLUSION(S): Blastocyst biopsy may not add additional risk to neonatal outcomes when compared with a control group.

Expanded carrier screening and preimplantation genetic diagnosis in a couple who delivered a baby affected with congenital factor VII deficiency.

BACKGROUND: Preimplantation genetic diagnosis (PGD) is a powerful tool for preventing the transmission of Mendelian disorders from generation to generation. However, PGD only can identify monogenically inherited diseases, but not other potential monogenic pathologies. We aimed to use PGD to deliver a healthy baby without congenital FVII deficiency or other common Mendelian diseases in a couple in which both individuals carried a deleterious mutation in the F7 gene. METHODS: After both members of the couple were confirmed to be carriers of the F7 gene mutation by Sanger sequencing, expanded carrier screening (ECS) for 623 recessive inheritance diseases was performed to detect pathological mutations in other genes. PGD and preimplantational genetic screening (PGS) were employed to exclude monogenic disorders and aneuploidy for their embryos. RESULTS: ECS using targeted capture sequencing technology revealed that the couple carried the heterozygous disease-causative mutations c.3659C > T (p.Thr1220Ile) and c.3209G > A (p.Arg1070Gln) in the CFTR gene. After PGD and PGS, one of their embryos that was free of congenital FVII deficiency, cystic fibrosis (CF) and aneuploidy was transferred, resulting in the birth of a healthy 3200 g male infant. CONCLUSION: We successfully implemented PGD for congenital FVII deficiency and PGD after ECS to exclude CF for the first time to the best of our knowledge. Our work significantly improved the reproductive outcome for the couple and provides a clear example of the use of ECS combined with PGD to avoid the delivery of offspring affected not only by identified monogenically inherited diseases but also by other potential monogenic pathologies and aneuploidy.

Newly expressed proteins of mouse embryonic fibroblasts irradiated to be inactive.

It has been found that post-radiation mouse embryonic fibroblasts can well maintain the pluripotency in human embryonic stem cells. However, the molecular mechanism remains unclear. In the present study, the new protein expression profile of post-radiation mouse embryonic fibroblasts was analyzed by immobilized pH gradient 2-dimensional polyacrylamide gel electrophoresis. Image analysis following silver staining revealed (969+/-57) vs. (1085+/-107) spots from post-radiation mouse embryonic fibroblasts and pre-radiation ones, respectively. Some newly expressed proteins, which were only abundantly present after irradiation, were subjected to peptide mass fingerprint analysis and identified using MALDI-TOF-MS, SWISS-PROT database, and RT-PCR. Several of those proteins were preliminarily identified to participate in cytokine secretion, cell signal transduction, transcriptional regulation, and apoptosis, etc., which suggested that inactive post-radiation mouse embryonic fibroblasts expressed some new proteins that may underlie the molecular mechanisms to maintain the pluripotency in human embryonic stem cells.

[Diagnosing achondroplasia by single cell nested-PCR].

OBJECTIVE: To research on the reliability of diagnosing achondroplasia (ACH) on single cell level and to provide a basis for preimplantation genetic diagnosis(PGD). METHODS: The high-frequency mutation region G380R of fibroblast growth factor receptor 3(FGFR3) gene was amplified by nested-PCR with single lymphocyte and single blastomere. The products of PCR were digested by restriction enzyme Bfm I, then the digested products were detected by 10% polyacrylamida gel electrophoresis(PAGE). RESULTS: The amplification success rate, allele dropout rate and correct diagnosis rate of single lymphocyte's PCR were 90.4%, 8.2% and 91.8%,respectively. The amplification success rate of single blastomere was 75.4%. CONCLUSION: The diagnosis of ACH by single cell nested-PCR is comparatively stable and reliable.