Immunosurgical studies on cytological and cytogenetic toxicity analysis of rat blastocysts after in vivo exposure to cyclophosphamide
Abstract
AIM: To establish immunosurgery and indices of cytogenetic assessment for
blastocyst and its inner cell mass (ICM), and to evaluate the toxic effects after
in vivo exposure to cyclophosphamide.
METHODS: Modified immunosurgery was established by preparation of rabbit-anti-rat
spleen antiserum and induction of diluted rat mixed serum as complement. Pregnant
rats on d 3 of gestation were injected i.p. cyclophosphamide (10, 20, and 40
mg.kg-1). On d 4, immunosurgery was performed on rat blastocysts. The cell number
and the micronuclei of blastocyst and ICM were evaluated respectively.
RESULTS: In the cyclophosphamide-treated rats, decreases of cell number (35 +/-
3, 32 +/- 1, 30 +/- 1, and 14 +/- 2, 11 +/- 1, 9 +/- 2) and increases of
frequency of micronuclei (1.81%, 2.27%, 3.14%, and 2.53%, 2.98%, 4.75%) in
blastocysts and ICM were observed in a dose-related manner. The changes of
blastocyst were, however, not parallel to those of ICM which were more serious.
CONCLUSION: Modified immunosurgery, an objective and elegant technique, was used
on rat blastocysts. In vivo could cyclophosphamide injured ICM more than
blastocysts.
Keywords:
blastocyst and its inner cell mass (ICM), and to evaluate the toxic effects after
in vivo exposure to cyclophosphamide.
METHODS: Modified immunosurgery was established by preparation of rabbit-anti-rat
spleen antiserum and induction of diluted rat mixed serum as complement. Pregnant
rats on d 3 of gestation were injected i.p. cyclophosphamide (10, 20, and 40
mg.kg-1). On d 4, immunosurgery was performed on rat blastocysts. The cell number
and the micronuclei of blastocyst and ICM were evaluated respectively.
RESULTS: In the cyclophosphamide-treated rats, decreases of cell number (35 +/-
3, 32 +/- 1, 30 +/- 1, and 14 +/- 2, 11 +/- 1, 9 +/- 2) and increases of
frequency of micronuclei (1.81%, 2.27%, 3.14%, and 2.53%, 2.98%, 4.75%) in
blastocysts and ICM were observed in a dose-related manner. The changes of
blastocyst were, however, not parallel to those of ICM which were more serious.
CONCLUSION: Modified immunosurgery, an objective and elegant technique, was used
on rat blastocysts. In vivo could cyclophosphamide injured ICM more than
blastocysts.