Immunosurgical studies on cytological and cytogenetic toxicity analysis of rat blastocysts after in vivo exposure to cyclophosphamide.

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.