Age-related increase in aneuploidy and alteration of gene expression in mouse first polar bodies.

Purpose: To confirm that aneuploidy candidate genes are detectable in the first polar body (PB(1)) of MII oocytes and to investigate the age-dependent molecular changes in PB(1).

Methods: Aged (12-to 15-mo-old) and young (2-mo-old) mice were administered pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotrophin (hCG). MII oocytes were obtained and the first PB was removed.

Detection and quantification of maternal-effect gene transcripts in mouse second polar bodies: potential markers of embryo developmental competence.

Objective: To test the hypothesis that quantification of messenger RNAs originating from the second polar body (PB(2)) provides a noninvasive tool for assessing embryo quality.

Design: Prospective study.

Setting: Hospital-based academic research laboratory.

Follicle microenvironment-associated alterations in gene expression in the mouse oocyte and its polar body.

Objective: To determine whether the follicle environment modulates oocyte-specific gene transcript levels in cultured oocytes and polar bodies (PBs).

Design: Animal study.

Setting: Large academic research center.

Age-associated alteration of oocyte-specific gene expression in polar bodies: potential markers of oocyte competence.

Objective: To confirm that oocyte-specific messenger RNAs are detectable in the polar body (PB) of metaphase II (MII) oocytes and determine the effect of age on oocyte-specific transcript levels.

Design: Prospective study.

Setting: Hospital-based academic research laboratory.

[Single cell analysis of some deletion in dystrophin gene exons and gender determination by 3-plex nested PCR].

Objective: To set up a technique of single lymphocytes 3-plex nested PCR for dystrophin and SRY gene, and to evaluate the possibility of using this technique for preimplantation genetic diagnosis(PGD) of deleted Duchenne muscular dystrophy (DMD) with family history.

Methods: Fifty single lymphocytes of a normal male and fifty of a normal female were obtained for detecting dystrophin gene(exon 51, exon 19, exon 48) and SRY gene by 3-plex nested PCR.

Results: In the group of exon 51/exon 19/SRY, the amplification rates of exon 51, exon 19 and SRY in male were 96%, 94% and 94%; the amplification rates of exon 51 and exon19 in female were 94% and 94%, respectively.

Birth of healthy children after preimplantation diagnosis of beta-thalassemia.

Background: Clinical programs for preventing beta-thalassemia are presently based on prospective carrier screening and prenatal diagnosis. This paper report an achievement of a pregnancy with unaffected embryos using in vitro fertilization and embryo transfer (IVF-ET), in combination with preimplantation genetic diagnosis (PGD), for a couple at risk of having children with beta-thalassemia.

Methods: A couple carrying different thalassemia mutations, both a codon 41 - 42 mutation and the IVS II 654 mutation, received standard IVF treatment, with intracytoplasmic sperm injection, embryo biopsiy, single cell polymerase chain reaction (PCR) and DNA analysis.

[Preimplantation genetic diagnosis for beta-thalassemia using whole genome amplification].

Objective: To achieve pregnancy with unaffected embryo using in vitro fertilization and embryo transfer (IVF-ET) and preimplantation genetic diagnosis(PGD) for the couples at risk of having children with beta-thalassemia.

Methods: A couple carrying different thalassemia mutations of codon 41/42 and codon IVS2 position 654 received standard IVF treatment and intracytoplasmic sperm injection, embryo biopsy, single cell polymerase chain reaction and DNA analyses, and only the unaffected or carrier embryos were transferred to uterus. Pregnancy confirmation, and prenatal diagnosis were done at 20 week's gestation.

[Successful preimplantation genetic diagnosis for beta-thalassemia using primer extension preamplification].

Objective: To achieve preimplantation genetic diagnosis (PGD) of the couples at risk of having children with beta-thalassemia, as an alternative to prenatal diagnosis.

Methods: Two couples carrying different thalassemia mutations of codon 41/42 and codon intervening sequence 2 position 654 received standard in vitro fertilization treatment and intracytoplasmic sperm injection, embryo biopsy and the whole genome was amplified by primer extension preamplification (PEP). Nested polymerase chain reaction was then used to amplify two mutation sites separately.

[Preimplantation genetic diagnosis for beta-thalassemia].

Objective: To investigate the effect of in vitro fertilization and embryo transfer (IVF-ET) and preimplantation genetic diagnosis (PGD) for the couples at risk of having children with beta-thalassemia.

Methods: Four couples carrying different thalassemia mutations received standard IVF treatment. Embryo biopsy was conducted.