Automated in Vitro Screening of Teratogens

• Published in: Toxicology and applied pharmacology 2002-05, Vol.181 (1), p.1-15
• Main Authors: Walmod, Peter S., Berezin, Anton, Gallagher, Helen C., Gravemann, Ute, Lepekhin, Eugene A., Belman, Vadym, Bacon, Christopher L., Nau, Heinz, Regan, Ciaran M., Berezin, Vladimir, Bock, Elisabeth
• Format: Article
• Language: English
• Subjects: AM580; Animal Testing Alternatives - instrumentation; Animal Testing Alternatives - methods; Animals; Biological and medical sciences; Cell Division - drug effects; Cell Line; cell morphology; Cell Survival - drug effects; developmental toxicity; Dose-Response Relationship, Drug; EM12; Embryology: invertebrates and vertebrates. Teratology; Fibroblasts - drug effects; Fibroblasts - pathology; Fundamental and applied biological sciences. Psychology; General aspects. Methods; high-throughput screening; image analysis; Image Processing, Computer-Assisted; Medical sciences; Mice; Microscopy, Video - instrumentation; Microscopy, Video - methods; phytanic acid; retinoic acid; teratogenicity; Teratogens - classification; Teratogens - toxicity; Teratology. Teratogens; thalidomide; Toxicology; valproic acid
• ISSN: 0041-008X; 1096-0333
• DOI: 10.1006/taap.2002.9393

We present a new in vitro assay for screening of potential teratogens, based on staining of cultured mouse fibroblastoid L929 cells for the determination of number of live and dead cells and of cell morphology, employing automatic video recording, followed by detection of the stained specimen and calculation of endpoint values by the use of a computerized microscope workstation. Ten different parameters were combined empirically into a single index describing general alterations in cell morphology, and, subsequently, measurements of alterations in morphology and proliferation were combined to produce a single empirical index aimed at predicting teratogenic potency. The assay was employed in two different laboratories on 10 coded compounds; 7 compounds that have demonstrated in vivo teratogenic potentials: valproic acid (VPA), pentyl-4-yn-VPA, retinoic acid (RA), 13-cis-RA, AM580, thalidomide, and α-EM12 and 3 compounds for which no teratogenic potential has been demonstrated: isobutyl-4-yn-VPA, phytanic acid, and β-EM12. Within each of the three groups of compounds the nonteratogens generally caused smaller alterations in cell morphology than the teratogens, although the effects of thalidomide and related compounds generally were minor or insignificant. The data support the hypothesis that cell morphology and proliferation in combination with other endpoints may be employed for in vitro screenings of potential teratogens, although studies of additional compounds are needed in order to establish the general validity of the procedure.