Postimplantation whole embryo culture assay for hamsters: an alternative to rat and mouse.

Bogdan Wlodarczyk,Bogumil Biernacki,Jan Zmudzki,Maria Minta

doi:10.1100/tsw.2001.48

Bogdan Wlodarczyk, Bogumil Biernacki + Show 2 more

Open Access

https://doi.org/10.1100/tsw.2001.48

Copy DOI

Abstract

Postimplantation whole embryo culture (WEC) assay for rats and mice has been well established and introduced to many laboratories. Recently WEC technique for rabbits has been developed; however, information on culture of other species is very limited. Knowing the usefulness of hamsters in classical embryotoxicology, we reasoned that hamster WEC could be an alternative model for the most frequently used rat and mouse WEC. Previously we have optimized culture conditions for postimplantation hamster embryos. The aim of this study was to test the susceptibility of hamster embryos cultured in vitro to embryotoxic compounds and to compare our results with those reported by others on rat or mouse embryo culture. For that purpose we choose three known embryotoxic compounds--valproic acid, cadmium chloride, and diethylstilbestrol--and tested them using a postimplantation hamster whole embryo culture assay. Hamster embryos were cultured from 7.5 days gestation for 24 h in a medium consisting of 35% hamster serum and 65% synthetic culture medium (Iscove's or McCoy 5A). At the end of the culture period, the embryos were examined morphologically, measured with the aid of a computer image analysis system, and total protein content was assessed. All three compounds exhibited dose-related embryotoxic and teratogenic effects in hamster embryos. The malformations observed were similar to those reported on rat and mouse embryos. Comparison of the results with data reported by other authors indicates that hamster embryos cultured in vitro might be more susceptible to embryotoxic stimuli than rat and mouse embryos.

Highlights

Rodent whole embryo culture (WEC), developed originally by New and his associates [1] in the 1960s, is used in many laboratories worldwide for evaluation of the mechanisms of development and teratogenesis
The whole embryo culture is frequently applied as a pre-screen test to detect embryotoxic potential of new chemical compounds, even though the final results of validation studies of WEC founded by the European Center for the Validation of Alternative Methods are not yet published
We considered whether the hamster might provide a better alternative model to the rat or mouse whole embryo culture

Summary

Introduction

Rodent whole embryo culture (WEC), developed originally by New and his associates [1] in the 1960s, is used in many laboratories worldwide for evaluation of the mechanisms of development and teratogenesis. The whole embryo culture is frequently applied as a pre-screen test to detect embryotoxic potential of new chemical compounds, even though the final results of validation studies of WEC founded by the European Center for the Validation of Alternative Methods are not yet published. The main advantages of this species include the shortest period of gestation (16 days) found among rodent laboratory animals, a fast progression through organogenesis, and a high susceptibility to teratogens in vivo. Given these advantages, we considered whether the hamster might provide a better alternative model to the rat or mouse whole embryo culture. There were big variations in size and development of the embryos as well as high incidence of malformations [7]

Objectives

Methods

Results