A simplified biopsy method for precompacted mouse embryos: A technical report

Abstract

This article presents a new, simple and rapid embryo biopsy method. The blastomere for genetic analysis can be separated from a precompacted mouse embryo after a partial zona digestion with the use of a holding pipette. For the micromanipulation only two microcapillaries and micromanipulators are needed. The development of the biopsied embryos was studied during in vitro culture and in utero following embryo transfer. There was no significant difference between the treated and the control groups in the ratio of embryos that developed to the blastocyst stage, although the biopsied embryos were delayed in their development because they contained significantly fewer cells compared to the control ones at the same stage. Although there was no difference in the ratio of implantation, the development of the biopsied embryos in utero was also delayed 12-24 hours on the 9th day of pregnancy. No difference in development was visible from the 13th day of pregnancy. Statistically, no differences were found in the developmental ratio (number of developed fetuses/transferred embryos) of the control and treated embryos during gastrulation (9th day of pregnancy), at the beginning of organogenesis (13th day of pregnancy) and before birth (19th day of pregnancy). The embryo biopsy method presented here can be a new and useful tool for preimplantation genetic diagnosis.

Vitrification of biopsied mouse embryos

“…Eight-cell stage embryos were biopsied as described earlier by Bodó et al (2002). The embryos were pre-treated in Ca 2+ -and Mg…”

“…The biopsy procedure itself has been shown to have no negative effect on the developmental potential of mouse embryos either when the zona was punctured (Wilton and Trounson, 1989;Krzyminska et al, 1990) or drilled (Garrisi et al, 1992;Bodó et al, 2002).…”