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For X/Y cell sortings by means of a FACS (B/D), we have been using Hoechst33342
(method Johnson et al.). After optimalisation of the staining method (time,
temp, H33342 conc), we found that it was possible to sort the H33342 stained
cells. But when using them for IVP of bovine embryo's, the cleavage rate was
slightly affected in a negative way and the further embryo development to the
blastocyst stage was dramatically affected in a negative way. Presumably, a lot
of groups are involved in this sort of semen sexing technique. As far as we
know, we are the first who wants to communicate freely about the observed
possible negative side effects of H33342 on In-Vitro embryo development.
Does someone has the same sort of experience with H33342 or other DNA staining
dyes within semen sorting trials and IVP? Is there more information available
or references about possible cytotoxic/ mutagenic effects of H33342 with or
without the interaction of UV laserlight ? In what way does H33342 act on the
integrity of the sperm DNA ? Does H33342 with or without laserlight give rise to
the formation of Reactive Oxygen Species ? Feel free to contact us directly or
by means of this mailing group. we will be glad in sharing research data on this
item.
Holland Genetics, the netherlands
Rene Haring, R & D Team
haring@nrs.nl
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