Assisted reproductive research: Laser assisted hatching and spindle detection (spindle view technique)

Abstract

Animal experiments are very important for the development of new assisted reproductive techniques (ART) for use in human and animal reproductive medicine. Most technical aspects of reproductive manipulation of humans and animals are very similar, and many components of successful human ART used nowadays have been derived from animal studies. In this study we examined (1) the use of 'non-contact' laser for assisted hatching, (2) whether spindles in living mouse oocytes could safely be imaged/examined by polarisation microscope (polscope) and (3) the influence of environment (e.g. temperature, in vitro culture, etc.) on spindle detection/visualisation. The data of the study presented here show that (1) laser assisted hatching (AH) is a fast, very accurate and safe procedure without any harmful effect on embryo development and it can support very effectively the implantation of embryos, (2) the use of polscope facilitates the evaluation of oocyte quality and the selection of oocytes with spindle, (3) by monitoring the spindle position during intracytoplasmic sperm injection (ICSI), we can reduce spindle damage and increase the chance of fertilisation. Further studies are underway to test the hypothesised connection between spindle birefringence and developmental capacity of oocytes/embryos.

Oocyte cryopreservation: the birth of the first Hungarian babies from frozen oocytes

“…In vitro fertilization with ICSI was performed 3-4 h after thawing, and fertilization was assessed 12-16 h later. Assisted hatching (AH) with laser was performed on all embryos prior to transfer [13]. Embryo transfer was carried out 48-72 h after ICSI at the 2 to 8 cell stages.…”

Deliveries from embryos fertilized with spermatozoa obtained from cryopreserved testicular tissue

“…Up to three embryos were selected for transfer into the uterine cavity. Prior to ET laser assisted hatching was performed on each of the embryos (MTG, Fertilase, Hamburg, Germany) [11]. Pregnancy was defined as a spontaneous rise in a βhCG concentration at least 10 days post-transfer.…”