The amido black assay: a simple and quantitative multipurpose test of adhesion, proliferation, and cytotoxicity in microplate cultures of keratinocytes (HaCaT) and other cell types growing adherently or in suspension

Schulz, Jörn; Dettlaff, Sabine; Fritzsche, Ute; Harms, Ute; Schiebel, Heike; Derer, Wolfgang; Fusenig, Norbert E.; Hülsen, Andrea; Böhm, Markus

doi:10.1016/0022-1759(94)90069-8

Cited by 53 publications

(28 citation statements)

References 26 publications

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“…Two different assays were used for determining the proliferation of stromal cells: 1) the MTT assay [18,19], and 2) the naphtol blue black assay [20]. Both assays demonstrated a linear correlation with cell number.…”

Section: Proliferation Assaysmentioning

confidence: 99%

Cell lineage characteristics of human prostatic stromal cells cultured in vitro

et al. 2000

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Section: Proliferation Assaysmentioning

confidence: 99%

Cell lineage characteristics of human prostatic stromal cells cultured in vitro

et al. 2000

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“…12 Various microplate colorimetric methods have been developed for assessing (i) viability and (ii) proliferation of cells. These include (i) neutral red, 13 trypan blue, 14 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT), 15 ethidium bromide, 16 and (ii) methylene blue, 17 amido black, 18 crystal violet, 19 acid phosphatase, 20 Hoechst dye, 21 etc. In our lab, some microplate cell culture methods have been modified to fit the biocompatibility testing purpose.…”

Section: Discussionmentioning

confidence: 99%

False positive results in cytotoxicity testing due to unexpectedly volatile compounds

Ciapetti

Granchi

Verri

et al. 1998

J. Biomed. Mater. Res.

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We investigated the cytotoxicity of different dental materials according to the study protocol adopted by our lab for the screening of new materials. Experimental parameters used in such testing are addressed mainly in documents EN 30993 "Biological evaluation of medical devices, Part 5: Tests for cytotoxicity: in vitro methods" and "Biological evaluation of medical devices, Part 12: Sample preparation and reference materials." Cells were cultured in microplates and challenged with aqueous extracts of the materials. The assay methods were neutral red- and propidium iodide-uptake assays, both indicative of cell viability and able to provide quantitative data. The observation of contrasting results for one material using the above-mentioned methods raised some concern about the assay system used. With further experimentation, it appeared that a sustained release of volatile substances still present in one extract exerted a toxic effect in neighboring cultures. It is concluded that in the microenvironment of a microplate the distribution of samples cannot be disregarded, as it may be responsible for toxicity cross-contamination. Moreover, the use of more than one single method has to be recommended in cytotoxicity testing, in order to avoid false positive results due to experimental artifacts.

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“…Amido black assay [13] : Cells were cultured on 96-well plates. Different concentrations of H17 were incubated for 24 h and 72 h. Then the medium was removed, cells were fixed by formaldehyde and, after aspiration of fixative and non-adherent cells, were stained by amido black at pH 3.5.…”

Section: Cytotoxicity Testingmentioning

confidence: 99%