title: Automated in vitro screening of teratogens.
abstract: 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 alpha-EM12 and 3 compounds for which no teratogenic potential has been demonstrated: isobutyl-4-yn-VPA, phytanic acid, and beta-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.
authors.affiliation: Protein Laboratory, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Peter S
authors.initials: PS
authors.lastname: Walmod
authors.forename: Anton
authors.initials: A
authors.lastname: Berezin
authors.forename: Helen C
authors.initials: HC
authors.lastname: Gallagher
authors.forename: Ute
authors.initials: U
authors.lastname: Gravemann
authors.forename: Eugene A
authors.initials: EA
authors.lastname: Lepekhin
authors.forename: Vadym
authors.initials: V
authors.lastname: Belman
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Bacon
authors.forename: Heinz
authors.initials: H
authors.lastname: Nau
authors.forename: Ciaran M
authors.initials: CM
authors.lastname: Regan
authors.forename: Vladimir
authors.initials: V
authors.lastname: Berezin
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Bock
mesh_terms: D000826:Animal Testing Alternatives / Q000295:instrumentation / Q000379:methods*; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects / Q000473:pathology; D007091:Image Processing, Computer-Assisted; D051379:Mice; D018715:Microscopy, Video / Q000295:instrumentation / Q000379:methods; D013723:Teratogens / Q000145:classification / Q000633:toxicity*
chemical_list: D013723:Teratogens