Three-dimensional imaging and quantification of mouse ovarian follicles via optical coherence tomography.

Ovarian tissue cryopreservation has been successfully applied worldwide for fertility preservation. Correctly selecting the ovarian tissue with high follicle loading for freezing and reimplantation increases the likelihood of restoring ovarian function, but it is a challenging process. In this work, we explore the use of three-dimensional spectral-domain optical coherence tomography (SD-OCT) to identify different follicular stages, compare the identifications with H&E images, and measure the size and age-related follicular density distribution differences in mice ovaries.

Three-dimensional imaging and quantification of mouse ovarian follicles via optical coherence tomography.

Ovarian tissue cryopreservation has been successfully applied worldwide for fertility preservation. Correctly selecting the ovarian tissue with high follicle loading for freezing and reimplantation increases the likelihood of restoring ovarian function, but it is a challenging process. In this work, we explore the use of three-dimensional spectral-domain optical coherence tomography (SD-OCT) to identify different follicular stages, especially primary follicles, compare the identifications with H&E images, and measure the size and age-related follicular density distribution differences in mice ovaries.

Chromatin Accessibility Analysis from Fresh and Cryopreserved Human Ovarian Follicles.

Understanding how gene regulatory elements influence ovarian follicle development has important implications in clinically relevant settings. This includes understanding decreased fertility with age and understanding the short-lived graft function commonly observed after ovarian tissue cryopreservation and subsequent autologous transplantation as a fertility preservation treatment. The Assay for Transposase Accessible Chromatin by sequencing (ATAC-seq) is a powerful tool to identify distal and proximal regulatory elements important for activity-dependent gene regulation and hormonal and environmental responses such as those involved in germ cell maturation and human fertility.

Potential roles of experimental reproductive technologies in infertile women with diminished ovarian reserve.

In assisted reproductive technology treatment, diminished ovarian reserve (DOR) is a condition of utmost clinical and scientific relevance because of its negative influence on patient outcomes. The current methods of infertility treatment may be unsuitable for many women with DOR, which support the need for development of additional approaches to achieve fertility restoration. Various techniques have been tried to improve the quality and increase the quantity of oocytes in DOR patients, including mitochondrial transfer, activation of primordial follicles, in vitro culture of follicles, and regeneration of oocytes from various stem cells.

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.

Birth of healthy children after preimplantation diagnosis of beta-thalassemia by whole-genome amplification.

Preimplantation genetic diagnosis (PGD) offers couples at risk for transmitting an inherited disorder the possibility to avoid the need to terminate affected pregnancies. PGD for monogenic diseases is most commonly accomplished by blastomere biopsy from cleavage-stage embryos, followed by PCR-based DNA analysis. However, the molecular heterogeneity of many monogenic diseases requires a diagnostic strategy capable of detecting a range of mutations and compound genotypes.

[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.

[Influence of advanced age on the outcome of in vitro fertilization and embryo transfer].

Objective: To determine the influence of age on the outcome of In vitro fertilization and embryo transfer (IVF-ET).

Methods: A retrospective study of 139 cycles of conventional IVF and 69 intracytoplasmic sperm injection (ICSI) cycles was performed between January 1999 and December 1999.

Results: A total of 208 patients (age 36 to 45 years) after IVF or ICSI were divided into five age groups (36 years group, n = 78; 37 years group, n = 49; 38 years group, n = 50; 39 years group, n = 18; 40 approximately 45 years group, n = 13).

[Preimplantation genetic diagnosis].

Preimplantation genetic diagnosis is a very early form of prenatal diagnosis aimed at eliminating embryos carrying serious genetic diseases before implantation. The basic techniques currently used involve embryo biopsy, the polymerase chain reaction and fluorescence in situ hybridization. In the current review, a number of problems arising from the use of these technologies as well as the possible solutions and new developments are discussed.