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BioAssay: AID 504640

A Cell Based Secondary Assay to Explore Cytotoxicity in THP-1 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without Glycerol

Project Overview: This functional assay was developed for the detection of compounds inhibiting THP-1 cell viability as a secondary screen to the M. tuberculosis bacteriocidal assay that compares growth in media containing glycerol versus growth in media without glycerol. The THP-1 cell line was chosen as a representative peripheral blood monocyte. ..more
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 Tested Compounds
 Tested Compounds
All(40)
 
 
Active(29)
 
 
Inactive(11)
 
 
 Tested Substances
 Tested Substances
All(40)
 
 
Active(29)
 
 
Inactive(11)
 
 
 Related BioAssays
 Related BioAssays
AID: 504640
Data Source: Southern Research Specialized Biocontainment Screening Center (TB_Inh_THP1_1)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-04-06
Hold-until Date: 2012-03-22
Modify Date: 2012-03-22

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 29
Related Experiments
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AIDNameTypeComment
435019A Cell Based Secondary Assay To Explore Cytotoxicity of Compounds that Inhibit Mycobacterium TuberculosisConfirmatorydepositor-specified cross reference
449762High Throughput Screening Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in 7H9 MediaConfirmatorydepositor-specified cross reference
449764A High Throughput Confirmatory Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in the absence of GlycerolConfirmatorydepositor-specified cross reference
449775Summary of Assays used to Identify Novel Compounds That Inhibit Mycobacterium Tuberculosis in 7H9 Media with GlycerolSummarydepositor-specified cross reference
493181A High Throughput Confirmatory Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in the absence of Glycerol using Purified and Synthesized CompoundsConfirmatorydepositor-specified cross reference
493198High Throughput Screening Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in 7H9 Media using Purified and Synthesized CompoundsConfirmatorydepositor-specified cross reference
504335A Cell Based Secondary Assay To Explore Cytotoxicity of Purified and Synthesized Compounds that Inhibit Mycobacterium Tuberculosis.Confirmatorydepositor-specified cross reference
504556High Throughput Screening Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in 7H9 Media using Purified and Synthesized Compounds (2)Confirmatorydepositor-specified cross reference
504564A High Throughput Confirmatory Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in the absence of Glycerol using Purified and Synthesized Compounds (2)Confirmatorydepositor-specified cross reference
504683A Cell Based Secondary Assay to Explore Cytotoxicity in THP-1 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without Glycerol (2)Confirmatorydepositor-specified cross reference
504684A Cell Based Secondary Assay To Explore Vero Cell Cytotoxicity of Purified and Synthesized Compounds that Inhibit Mycobacterium Tuberculosis (3)Confirmatorydepositor-specified cross reference
504852A Cell Based Secondary Assay to Explore Cytotoxicity in THP-1 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without Glycerol (3)Confirmatorydepositor-specified cross reference
504853A Cell Based Secondary Assay to Explore Cytotoxicity in HepG2 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without Glycerol (3)Confirmatorydepositor-specified cross reference
504854A Cell Based Secondary Assay To Explore Vero Cell Cytotoxicity of Purified and Synthesized Compounds that Inhibit Mycobacterium Tuberculosis (4)Confirmatorydepositor-specified cross reference
504857High Throughput Screening Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in 7H9 Media using Purified and Synthesized Compounds (4)Confirmatorydepositor-specified cross reference
504860A High Throughput Confirmatory Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in the absence of Glycerol using Purified and Synthesized Compounds (4)Confirmatorydepositor-specified cross reference
504897High Throughput Screening Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in 7H9 Media using Purified and Synthesized Compounds (6)Confirmatorydepositor-specified cross reference
504898A High Throughput Confirmatory Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in the absence of Glycerol using Purified and Synthesized Compounds (5)Confirmatorydepositor-specified cross reference
504901A High Throughput Confirmatory Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in the absence of Glycerol using Purified and Synthesized Compounds (6)Confirmatorydepositor-specified cross reference
504903High Throughput Screening Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in 7H9 Media using Purified and Synthesized Compounds (5)Confirmatorydepositor-specified cross reference
504909A Cell Based Secondary Assay To Explore Vero Cell Cytotoxicity of Purified and Synthesized Compounds that Inhibit Mycobacterium Tuberculosis (5)Confirmatorydepositor-specified cross reference
504910A Cell Based Secondary Assay to Explore Cytotoxicity in HepG2 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without Glycerol (4)Confirmatorydepositor-specified cross reference
504911A Cell Based Secondary Assay to Explore Cytotoxicity in THP-1 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without Glycerol (4)Confirmatorydepositor-specified cross reference
588437A High Throughput Confirmatory Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in the absence of Glycerol using Purified and Synthesized Compounds (7)Confirmatorydepositor-specified cross reference
588441A Cell Based Secondary Assay to Explore Cytotoxicity in HepG2 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without Glycerol (5)Confirmatorydepositor-specified cross reference
588443A Cell Based Secondary Assay to Explore Cytotoxicity in THP-1 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without Glycerol (5)Confirmatorydepositor-specified cross reference
588445A Cell Based Secondary Assay To Explore Vero Cell Cytotoxicity of Purified and Synthesized Compounds that Inhibit Mycobacterium Tuberculosis (6)Confirmatorydepositor-specified cross reference
588447High Throughput Screening Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in 7H9 Media using Purified and Synthesized Compounds (8)Confirmatorydepositor-specified cross reference
602431A Cell Based Secondary Assay To Explore Vero Cell Cytotoxicity of Purified and Synthesized Compounds that Inhibit Mycobacterium Tuberculosis (8)Confirmatorydepositor-specified cross reference
602432A Cell Based Secondary Assay to Explore Cytotoxicity in HepG2 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without Glycerol (6)Confirmatorydepositor-specified cross reference
602433A Cell Based Secondary Assay to Explore Cytotoxicity in THP-1 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without Glycerol (6)Confirmatorydepositor-specified cross reference
602435High Throughput Screening Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in 7H9 Media using Purified and Synthesized Compounds (9)Confirmatorydepositor-specified cross reference
602437A High Throughput Confirmatory Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in the absence of Glycerol using Purified and Synthesized Compounds (8)Confirmatorydepositor-specified cross reference
504562A Cell Based Secondary Assay To Explore Cytotoxicity of Purified and Synthesized Compounds that Inhibit Mycobacterium Tuberculosis (2)Confirmatorysame project related to Summary assay
504642A Cell Based Secondary Assay to Explore Cytotoxicity in HepG2 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without GlycerolConfirmatorysame project related to Summary assay
504645A High Throughput Confirmatory Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in the absence of Glycerol using Purified and Synthesized Compounds (3)Confirmatorysame project related to Summary assay
504646High Throughput Screening Assay used to Identify Novel Compounds that Inhibit Mycobacterium Tuberculosis in 7H9 Media using Purified and Synthesized Compounds (3)Confirmatorysame project related to Summary assay
504682A Cell Based Secondary Assay to Explore Cytotoxicity in HepG2 Cells of Compounds that Modulate Mycobacterium tuberculosis in Media Containing Glycerol versus Media without Glycerol (2)Confirmatorysame project related to Summary assay
Description:
Southern Research's Specialized Biocontainment Screening Center (SRSBSC)
Southern Research Institute (Birmingham, Alabama)
NIH Molecular Libraries Probe Production Centers Network (MLPCN)
Assay Provider: William Bishai, Johns Hopkins University
Award: 1 R03 MH084877-01A1

Project Overview: This functional assay was developed for the detection of compounds inhibiting THP-1 cell viability as a secondary screen to the M. tuberculosis bacteriocidal assay that compares growth in media containing glycerol versus growth in media without glycerol. The THP-1 cell line was chosen as a representative peripheral blood monocyte.

In this assay, we treated THP-1 cells with compounds selected as "hits" in the comparison with and without glycerol M. tuberculosis assay (AID 449762) for 72 hours over a 10 point 2-fold dilution series, ranging from 40uM to 0.078uM. Following 72 hours of treatment, relative viable cell number was determined using Cell Titer Glo from Promega. Each plate contained 64 replicates of vehicle treated cells which served as negative controls.
Protocol
Cell Culture: THP-1 cells were subcultured every 7 days in RPMI 1640 with 10% fetal bovine serum, incubated at 37 degrees C in 5% carbon dioxide. Cells were passaged as needed, harvested from flasks using 0.25% trypsin-EDTA and maintained for no more than 20 passages.

Compound Dosing/Plating: Compounds and carrier controls were diluted in complete growth medium to prepare a 6X concentrated dosing solution which was dispensed into 384-well black clear-bottom tissue culture treated plates (5 uL volume).

Cell Plating: Twenty uL of complete growth medium containing 3000 cells were dispensed per well. Plates were incubated at 37 C, 5% CO2 for 72h prior to endpoint detection.

Endpoint/Detection: At the end of the treatment period, assay plates were removed from the incubator and equilibrated to room temperature for 10 min. Twenty-five muL of Cell Titer Glo reagent was added and plates were incubated for an additional 10 min in the dark. At the end of the incubation, assay plates were analyzed using a PerkinElmer Envision microplate reader in luminescence mode with an integration time of 0.1 s.

Data Analysis: Sixty-four control wells containing cells treated with DMSO vehicle were included on each assay plate. Compound data was normalized and reported as % viability which was calculated using the following formula: % viability = 100*(Cmpd Lum-Med background)/(Med Cell Ctrl - Med background). The normalized % viability was plotted against the tested concentrations. The CC50 values were calculated using XLfit formula 205, a 4 parameter Levenburg-Marquardt algorithm with maximum and minimum limits set at 100 and 0.
Comment
Outcome: Compounds that showed <70% cell viability for at least one concentration were defined as "Active". If the % viability at all doses was >70%, the compound was defined as "Inactive".

Scoring: SRBCSC uses a three-tier scoring system where scores of 0-40 apply to primary screen data, 41-80 indicates confirmatory screen data, and 81-100 is reserved for confirmatory data on resynthesized compounds. Inactive compounds at any screening level receive a score of 0. In this confirmatory screen, "Active" compounds were scored based on CC50 results on a tier of 81-100 with "Inactive" compounds scoring 0.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Format: Cell-based
Assay Type: Toxicity
Assay Cell Type: THP-1
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1CC50 ModifierString
2CC50*Cytotoxic concentrationFloatμM
3CC50 Std Dev ModifierString
4CC50 Std DevFloat
5CC50 Hill SlopeFloat
6CC50 Normalized Chi2Float
7% Cell Viability @ 100 uM (100μM**)Float%
8% Cell Viability @ 50 uM (50μM**)Float%
9% Cell Viability @ 25 uM (25μM**)Float%
10% Cell Viability @ 12.5 uM (12.5μM**)Float%
11% Cell Viability @ 6.25 uM (6.25μM**)Float%
12% Cell Viability @ 3.125 uM (3.125μM**)Float%
13% Cell Viability @ 1.563 uM (1.563μM**)Float%
14% Cell Viability @ 0.781 uM (0.781μM**)Float%
15% Cell Viability @ 0.391 uM (0.391μM**)Float%
16% Cell Viability @ 0.195 uM (0.195μM**)Float%

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: 1 R03 MH084877-01A1

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
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