Ultra-fast vitrification and rapid elution of human oocytes: part I. germinal vesicle model validation.

Research Question: Can GV-oocytes serve as an effective model to test the efficacy of ultra-fast vitrification (UFV)/ rapid elution (RE) treatments to support reliable, high survival rates and sustained functionality?

Design: Prospective pilot cohort studies were performed to investigate the feasibility of non-equilibration, UFV to sustain cellular integrity and development in contrast to control vitrification (CV: 10-15min ES/ 1min VS). In Phase 1, we applied a 2 × 2 factorial design (n=25-30 eggs/group) to evaluate post-warming dilution treatments: conventional multi-step (CD) versus rapid elution (RE; one-step), including an apriori fresh egg control group. Phase 1/2 focused on survival and maturation assessments, including meiotic spindle formation (Phase 2).

Ultra-fast vitrification and rapid elution of human oocytes: Part II - verification of blastocyst development from mature oocytes.

Research Question: Will ultra-fast vitrification (UFV) and rapid elution of mature human oocytes retain the reliable, high survival rates and meiotic spindle normality seen in the germinal vesicle model, and will these oocytes maintain their developmental competence to form blastocyst-stage embryos following artificial oocyte activation (AOA)?

Design: Conventional vitrification treatment was compared with UFV treatment in mature, germinal-vesicle-derived oocytes (Phase 2, Expt. 2, n = 50) and substandard donor oocytes, metaphase I-metaphase II (MII) oocytes and poor-quality MII oocytes (n = 222). Post-warming survival, the integrity of the meiotic spindle and AOA-related development were assessed.

Use of federated learning to develop an artificial intelligence model predicting usable blastocyst formation from pre-ICSI oocyte images.

Research Question: Can federated learning be used to develop an artificial intelligence (AI) model for evaluating oocyte competence using two-dimensional images of denuded oocytes in metaphase II prior to intracytoplasmic sperm injection (ICSI)?

Results: The oocyte AI model demonstrated area under the curve (AUC) up to 0.65 on two blind test datasets. High sensitivity for predicting competent oocytes (83-88%) was offset by lower specificity (26-36%).

Cryopreservation and IVF in the time of Covid-19: what is the best good tissue practice (GTP)?

Examine good tissue practices as relates to in vitro fertilization, biopsying, and vitrificationto compare current knowledge of ova, sperm, and embryos as vectors for disease transmission as it relates to our current knowledge regarding the SARS-CoV-2 virus.Unknown risks relating to the SARS-CoV-2 virus and sperm, ova, and embryos necessitate a reexamining of how human IVF is performed. Over the last decade, improvements in cryosurvival and live birth outcomes have been associated with zona pellucida breaching procedures (e.

Clinical benefits of preimplantation genetic testing for aneuploidy (PGT-A) for all in vitro fertilization treatment cycles.

The clinical application of a PGT-A program implementing single euploid embryo transfer is evaluated over a 6.5 year period, beginning with its early validation phases. Euploidy embryo status is inversely correlated to oocyte source age and positively correlated to blastocyst quality grades.

Day 7 blastocyst euploidy supports routine implementation for cycles using preimplantation genetic testing.

Objective: To determine if Day 7 blastocysts merit biopsy, vitrification and transfer consideration by contrasting their aneuploidy and implantation rates to Day 5 and 6 blastocysts.

Methods: A total of 1,925 blastocysts were biopsied from 402 PGT-A cycles over a 12 to 16 month interval. All embryos were cultured under tri-gas, humidified conditions (37ºC) for up to 7 days (168 hours post-insemination).

Comprehensive assessment of cryogenic storage risk and quality management concerns: best practice guidelines for ART labs.

Recent publicized events of cryogenic storage tank failures have created nationwide concern among infertility patients and patients storing embryos and gametes for future use. To assure patient confidence, quality management (QM) plans applied by in vitro fertilization (IVF) laboratories need to include a more comprehensive focus on the cryostorage of reproductive specimens. The purpose of this review is to provide best practice guidelines for the cryogenic storage of sperm, oocytes, embryos, and other reproductive tissues (e.

Modified MicroSecure Vitrification: A Safe, Simple and Highly Effective Cryopreservation Procedure for Human Blastocysts.

Clinical embryo vitrification evolved with the development of unique vitrification devices in the 21 century and with the misconception that ultra-rapid cooling in an "open" system (i.e., direct LN2 contact) was a necessity to optimize vitrification success.

Single center validation of routine blastocyst biopsy implementation.

Purpose: The study aims to contrast the efficacy of trophectoderm biopsy preimplantation genetic screening (PGS)/vitrification (VTF)-all cycles to past treatment protocols. Specifically, do these applied technologies increase live birth rates on a per cycle/first transfer basis?

Materials And Methods: An observational, retrospective cohort study of first transfer outcomes was performed in two groups. Group 1 (PGS) included PGS/VTF-all cycles, and group 2 (no PGS) included the first transfer from non-PGS fresh cycles or VTF-ALL cycles.

Validation-verification of a highly effective, practical human testicular tissue in vitro culture-cryopreservation procedure aimed to optimize pre-freeze and post-thaw motility.

Purpose: The aim of our paper was to validate a testicular biopsy procedure that simplifies handling, processing, and cryopreservation, while at the same time optimizes sperm motility before freezing and after thawing.

Methods: Two prospective studies were conducted to verify, optimize, and understand the virtues of pre-freeze testicular tissue IVC at different temperatures (21, 30, or 37 °C). Testicular tissue was obtained from clinical specimens designated for whole tissue cryopreservation (i.

Validation of microSecure vitrification (μS-VTF) for the effective cryopreservation of human embryos and oocytes.

A novel, aseptic closed system vitrification (VTF) technique for the cryopreservation of embryos and oocytes has been developed and clinically validated in this study. It combines the practicality of embryo-containing sterile flexipettes stored safely and securely with 0.3 ml CBS™ embryo straws possessing weld seals.

Potential risk of monochorionic dizygotic twin blastocyst formation associated with early laser zona dissection of group cultured embryos.

Objective: To document the risk of in vitro monochorionic dizygotic twin formation in the implementation of a program of blastocyst biopsy with preimplantation genetic screening (PGS).

Design: Case report.

Setting: Private infertility laboratory.

Safety and efficacy of topical azithromycin ophthalmic solution 1.0% in the treatment of contact lens-related dry eye.

Purpose: The purpose of this pilot study was to evaluate the safety and efficacy of azithromycin ophthalmic solution 1% in patients with contact lens-related dry eye (CLDE).

Methods: This was a 4-week, single-center, open-label clinical trial in patients diagnosed with CLDE using the Contact Lens Dry Eye Questionnaire (CLDEQ). Fifty patients were enrolled in this study.

A putative mesenchymal stem cells population isolated from adult human testes.

Mesenchymal stem cells (MSCs) isolated from several adult human tissues are reported to be a promising tool for regenerative medicine. In order to broaden the array of tools for therapeutic application, we isolated a new population of cells from adult human testis termed gonadal stem cells (GSCs). GSCs express CD105, CD166, CD73, CD90, STRO-1 and lack hematopoietic markers CD34, CD45, and HLA-DR which are characteristic identifiers of MSCs.

An effective, simplified, and practical approach to intracytoplasmic sperm injection at multiple IVF centers.

Purpose: Our purpose was to use effectively a simplified approach for intracytoplasmic sperm injection (ICSI) that achieves high levels of fertilization, embryo cleavage, and subsequent establishment of clinical pregnancies.

Methods: In a multicenter clinical analysis, 1814 mature oocytes were injected with a single sperm using a finely beveled (30 degrees) pipette without a spiked tip that was backfilled with an inert, silicone polymer. The injection system was composed of a sterile water-filled tubing interconnecting the pipette and a 20-gauge needle attached to a three-way stopcock and 5-ml disposable syringe.

Intracytoplasmic sperm injection and embryo development of human oocytes cryopreserved using 1,2-propanediol.

This study reports the subsequent embryo development of cryopreserved mature human oocytes following insemination or intracytoplasmic sperm injection (ICSI). Metaphase II oocytes were cryopreserved using a slow freezing-rapid thawing procedure employing the cryoprotectant 1,2-propanediol. The study was conducted at two centres.

Enzymatic amplification of specific deoxyribonucleic acid sequences from single cells: evaluation of a simplified and rapid method for use in preimplantation genetic diagnosis.

Objective: To develop a simplified polymerase chain reaction (PCR) protocol on single cells for the purpose of preimplantation genetic diagnosis. Also to evaluate a new thermal cycler, RoboCycler 40 (Stratagene, La Jolla, CA), for reducing the time to complete PCR amplification.

Design: PCR amplification without DNA purification or reamplification of a 149 base pair (bp) segment of the human Y chromosome was used as a model.

Developmental competence of mouse embryos following zona drilling using a non-contact holmium:yttrium scandian gallium garnet (Ho:YSGG) laser system.

The purpose of this study was to assess the efficacy of the holmium:yttrium scandian gallium garnet (Ho:YSGG) laser, operating in a pipette-free, non-contact mode, to assist hatching and sustain normal embryonic development. Two-cell mouse embryos were recovered and assigned to laser-assisted hatching (LAH) treatment or control human tubal fluid (HTF) culture with or without serum (HTF-s, HTF-o) or with late serum supplementation (HTF-o/s). The basic experimental apparatus for LAH consisted of a stationary 2.

Assisted hatching in mouse embryos using a noncontact Ho:YSGG laser system.

Purpose: A noncontact holmium:yttrium scandium gallium garnet (Ho:YSGG) laser system has been designed and tested for the micromanipulation of mammalian embryos. The purpose of this preliminary investigation was to determine the effectiveness of this laser for assisted hatching and evaluate its impact on embryo viability. The Ho:YSGG system, utilizing 250-microsecond pulses at a wavelength of 2.

Physiological characterization of blastocyst hatching mechanisms by use of a mouse antihatching model.

Objectives: To use the protein-free medium blastocyst antihatching model to characterize physiological events that mediate hatching.

Design: In a series of four prospective experiments, our aims were to [1] test the efficacy of the antihatching model and assisted hatching; [2] exam the influence of initial in vivo developmental stage and late serum supplementation on hatching inhibition; [3] discount the role of zona hardness and physical expansion directly affecting hatching; and [4] provide evidence that the trophectoderm is directly responsible for secreting a zona lysin.

Setting: University-based research laboratory.

Enzymatic characterization of zona pellucida hardening in human eggs and embryos.

Purpose: To characterize possible hardening of the human zona pellucida (ZP) and evaluate the effect of culture duration, patient age, and ZP thickness, ZP of unfertilized eggs (experiment 1, n = 367; experiment 2, n = 174) and abnormal embryos (experiment 1, n = 52) were randomly designated for alpha-chymotrypsin treatment after 0, 24, 48, 72, 96, 120 h (experiment 1) and 48 h, 72 h, and 1 week (experiment 2) of in vitro culture in HTF medium supplemented with 0.5% human serum albumin. Mean ZP thickness was predetermined in experiment 2.

Successful intrauterine insemination of Eld's deer (Cervus eldi thamin) with frozen-thawed spermatozoa.

This study tested the efficacy of assisted reproduction (synchronization of oestrus and intrauterine artificial insemination (AI)) in contributing to the captive propagation of an endangered species, the Eld's deer (Cervus eldi thamin). Semen was collected from males preselected on the basis of under-represented genotype. Motility of spermatozoa after thawing from ejaculates diluted with BF5F extender (8% glycerol), frozen on dry ice in 0.

Lack of antibody specificity by mouse trophectoderm during immunosurgery.

Immunosurgery of blastocyst-stage embryos is an effective procedure for isolating the inner cell mass. The ability of rabbit sera antibodies produced to interspecific types of cells to bind to mouse trophectoderm antigens and mediate complement-reactive lysis was investigated. Fifty hatched and 25 expanded blastocysts per treatment were exposed to 1 of 7 heat-inactivated polyclonal antibodies (1: 10 DMEM dilution) produced in rabbits to mouse brain cells (MBr), mouse lymphocytes (MLy), rat lymphocytes (RLy), hamster lymphocytes (HLy), cattle splenocytes (CSp), sheep splenocytes (SSp), and pig splenocytes (PSp).

Relationship of oestrus synchronization method, circulating hormones, luteinizing hormone and prostaglandin F-2 alpha receptors and luteal progesterone concentration to premature luteal regression in superovulated sheep.

Ewes were treated with exogenous follicle-stimulating hormone (FSH) and oestrus was synchronized using either a dual prostaglandin F-2 alpha (PGF-2 alpha) injection regimen or pessaries impregnated with medroxy progesterone acetate (MAP). Natural cycling ewes served as controls. After oestrus or AI (Day 0), corpora lutea (CL) were enucleated surgically from the left and right ovaries on Days 3 and 6, respectively.

Analysis of cryoprotectant, cooling rate and in situ dilution using conventional freezing or vitrification for cryopreserving sheep embryos.

Two studies were conducted to evaluate the influence of cryoprotectant, cooling rate, container and cryopreservation procedure on the post-thaw viability of sheep embryos. In Study 1, late morula- to blastocyst-stage embryos were exposed to 1 of 10 cryoprotectant (1.5 M, glycerol vs propylene glycol)-plunge temperature treatments.