New hyaluronan-based biomatrix for 3-D follicle culture yields functionally competent oocytes.

Background: Encapsulation of follicles within a biomatrix is one approach to maintaining 3-D follicle architecture during culture. Hyaluronan is one component of the natural extracellular matrix (ECM) that provides support to cells in vivo. This report describes the application of a novel tyramine-linked hyaluronan for 3-D in vitro follicle culture and the production of developmentally competent metaphase II oocytes.

Dynamic shapes of the zygote and two-cell mouse and human.

Mouse zygote morphokinetics were measured during interphase, the mitotic period, cytokinesis, and two-cell stage. Sequences of rounder-distorted-rounder shapes were revealed, as were changing patterns of cross section area. A calcium chelator and an actin-disrupting agent inhibited the area changes that occurred between pronuclear envelope breakdown and cytokinesis.

Delayed blastulation, multinucleation, and expansion grade are independently associated with live-birth rates in frozen blastocyst transfer cycles.

Objective: To identify blastocyst features independently predictive of successful pregnancy and live births with vitrified-warmed blastocysts.

Design: Retrospective study.

Setting: Academic hospital.

The new Rapid-i carrier is an effective system for human embryo vitrification at both the blastocyst and cleavage stage.

Background: The Rapid-i is a new FDA cleared closed carrier for embryo vitrification. The cooling rate of - 1220°C/min is far lower than that reported with open vitrification systems such as the cryoloop (-15,000°C/min). Little published data is currently available on this device.

Development of a xeno-free non-contact co-culture system for derivation and maintenance of embryonic stem cells using a novel human endometrial cell line.

Purpose: Mouse embryonic fibroblast feeder layers (MEF) have conventionally been used to culture and maintain the pluripotency of embryonic stem cells (ESC). This study explores the potential of using a novel human endometrial cell line to develop a non-xeno, non-contact co-culture system for ESC propagation and derivation. Such xeno-free systems may prove essential for the establishment of clinical grade human ESC lines suitable for therapeutic application.

Three dimensional culture of fresh and vitrified mouse pre-antral follicles in a hyaluronan-based hydrogel: a preliminary investigation of a novel biomaterial for in vitro follicle maturation.

Background: Folliculogenesis within the ovary requires interaction between somatic cell components and the oocyte. Maintenance of 3-dimensional (3-D) architecture and granulosa-oocyte interaction may be critical for successful in vitro maturation of follicles. Testing of novel biomaterials for the 3-D culture of follicles may ultimately lead to a culture model that can support the longer in vitro culture intervals needed for in vitro maturation of human oocytes from ovarian tissue biopsies.

Cryopreservation of individually selected sperm: methodology and case report of a clinical pregnancy.

Objective: To describe a new technique for freezing individually isolated spermatazoa from testicular biopsies, epididymal aspirates and oligospermic semen samples

Methods: Samples were evaluated for the presence of motile sperm before cryopreservation. Motile or twitching sperm were isolated with an ICSI needle for single sperm cryopreservation. Selected sperm were loaded on the High Security Straw (HSV; Irvine Scientific; Irvine,CA), in ~0.

Vitrification in open and closed carriers at different cell stages: assessment of embryo survival, development, DNA integrity and stability during vapor phase storage for transport.

Background: High cooling rates with vitrification can be achieved through the use of carriers that allow cryopreservation in fluid volumes < one μl. Open carriers allow direct contact of embryos with liquid nitrogen (LN2) whereas closed carrier systems sequester the embryo within a sealed system during immersion in LN2. The use of closed systems may be preferable to reduce the possibility of cross-contamination.

Three-dimensional in vitro follicle growth: overview of culture models, biomaterials, design parameters and future directions.

In vitro ovarian follicle culture is a new frontier in assisted reproductive technology with tremendous potential, especially for fertility preservation. Folliculogenesis within the ovary is a complex process requiring interaction between somatic cell components and the oocyte. Conventional two-dimensional culture on tissue culture substrata impedes spherical growth and preservation of the spatial arrangements between oocyte and surrounding granulosa cells.

Tranexamic acid treatment for heavy menstrual bleeding: a randomized controlled trial.

Objective: To assess the efficacy and safety of an oral formulation of tranexamic acid for the treatment of heavy menstrual bleeding.

Methods: Adult women with heavy menstrual bleeding (mean menstrual blood loss 80 mL or more per cycle) were enrolled in a double-blind, placebo-controlled study. After two pretreatment menstrual cycles, women were randomized to receive tranexamic acid 3.

Clinical pregnancy and live births after transfer of embryos vitrified on day 3.

Human embryo vitrification is a promising new technology but clinical outcome data is needed to gauge its effectiveness and safety. While pregnancy and live-birth data is available for blastocyst vitrification, such information is lacking for human embryo vitrification at the 6- to 8-cell stage. The current work presents clinical and obstetric outcomes from the transfer of embryos vitrified on day 3 at the cleavage stage.

Paternal effect on genomic activation, clinical pregnancy and live birth rate after ICSI with cryopreserved epididymal versus testicular spermatozoa.

Background: This study takes an in depth look at embryonic development, implantation, pregnancy and live birth rates with frozen epididymal and testicular sperm from obstructed (OA) and non-obstructed (NOA) patients.

Methods: Paternal effect of sperm source on zygote formation, embryonic cleavage, and genomic activation were examined. Additional outcome parameters monitored were clinical pregnancy rate (CPR), implantation rate (IR) and live birth rate.

Granulocyte-macrophage colony stimulating factor (GM-CSF) and co-culture can affect post-thaw development and apoptosis in cryopreserved embryos.

Purpose: The objective of this study was to evaluate the effects of growth factor supplementation and Vero cell co-culture on apoptosis and development of frozen thawed one-cell mouse embryos.

Methods: The following treatment regimens were assessed: (a) control medium (b) Vero cell co-culture and (c) growth factor supplemented medium. The individual growth factors tested were: GM-CSF, IGF-I, IGF-II, TNF-alpha, FGF-4, LIF, TGF-alpha, TGF-beta, IL-6, PDGF and EGF.