title: Morphometric and colorimetric analyses of human tumor cell line growth and drug sensitivity in soft agar culture.
abstract: Previous studies have demonstrated the suitability of image analysis of tetrazolium-stained colonies to assess growth and drug sensitivity of human tumor cells cultivated in soft agar culture. In the present study, the potential utility of colorimetric analysis to expedite experimental drug evaluations using human tumor cell lines was investigated. The same culture dishes were assessed by image analysis and by formazan colorimetry for purposes of comparing multiple methods of measuring growth as well as growth inhibition. Replicate cultures treated with 2-(p-iodonitrophenyl)-3-p-nitrophenyl-5-phenyltetrazolium chloride or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide exhibited nearly identical colony count and volume indices as well as excellent correlation in colorimetric end points. Colony-forming unit volume analysis versus colorimetric assessment of the same cultures following dimethyl sulfoxide extraction of protamine sulfate-rinsed, dried soft agar cultures exhibited excellent linear correlation for both growth (Pearson r ranging from 0.95 to 1.00) and drug sensitivity (Pearson r ranging from 0.90 to 0.99, and Spearman r ranging from 0.82 to 0.97) and similar drug sensitivity profiles. Results of the current investigation indicate that end points of soft agar culture remain stable for a period of at least 2 weeks following assay termination. In addition, a colorimetric detection range of 1.3-2.2 log units permits determinations of survival levels ranging from 100 to 5% of respective control levels. Colorimetric analysis is anticipated to expedite soft agar colony formation assay evaluations (a) by reducing the need to use the more rigorous and time-consuming image analysis procedures to measure activity in preliminary drug sensitivity assays and (b) by permitting the determination of effective concentration ranges of new experimental agents for subsequent, more detailed investigations.
authors.affiliation: Laboratory of Drug Discovery Research and Development, National Cancer Institute, Frederick, Maryland 21701-1013.
authors.forename: M C
authors.initials: MC
authors.lastname: Alley
authors.forename: C M
authors.initials: CM
authors.lastname: Pacula-Cox
authors.forename: M L
authors.initials: ML
authors.lastname: Hursey
authors.forename: L R
authors.initials: LR
authors.lastname: Rubinstein
authors.forename: M R
authors.initials: MR
authors.lastname: Boyd
mesh_terms: D001761:Bleomycin / Q000494:pharmacology; D002330:Carmustine / Q000494:pharmacology; D002455:Cell Division; D002460:Cell Line; D002699:Chlorambucil / Q000494:pharmacology; D002945:Cisplatin / Q000494:pharmacology; D003114:Colony-Forming Units Assay; D003124:Colorimetry; D003609:Dactinomycin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000494:pharmacology; D004361:Drug Tolerance; D005472:Fluorouracil / Q000494:pharmacology; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008627:Mercuric Chloride / Q000494:pharmacology; D016685:Mitomycin; D008937:Mitomycins / Q000494:pharmacology; D009369:Neoplasms / Q000473:pathology*; D013778:Tetrazolium Salts; D014747:Vinblastine / Q000494:pharmacology
chemical_list: D008937:Mitomycins; D013778:Tetrazolium Salts; D001761:Bleomycin; D002699:Chlorambucil; D003609:Dactinomycin; D016685:Mitomycin; D008627:Mercuric Chloride; D014747:Vinblastine; D004317:Doxorubicin; D002945:Cisplatin; D005472:Fluorouracil; D002330:Carmustine