Evidence map›Paper›PMID 29510509›Full record

ArticleInternational journal of molecular sciences2018

Miniaturization of the Clonogenic Assay Using Confluence Measurement.

Christian Mayr, Marlena Beyreis, Heidemarie Dobias, Martin Gaisberger, Martin Pichler, Markus Ritter, Martin Jakab, Daniel Neureiter, Tobias Kiesslich

Open access · goldAbstract read
In one paragraph

Article in International journal of molecular sciences, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.

0numbers the graph read from it
0cells of the map it votes in
6citing papers in PubMed
0.4field-weighted citation impact, top 35% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

6 citing papers in PubMed, 9 citations in OpenAlex.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

9 authors at 2 institutions in 2 countries.

Christian MayrLaboratory for Tumour Biology and Experimental Therapies (TREAT), Institute of Physiology and Pathophysiology, Paracelsus Medical University Salzburg, Strubergasse 22, 5020 Salzburg, Austria. christian.mayr@pmu.ac.at.
Marlena BeyreisLaboratory for Tumour Biology and Experimental Therapies (TREAT), Institute of Physiology and Pathophysiology, Paracelsus Medical University Salzburg, Strubergasse 22, 5020 Salzburg, Austria. marlena.beyreis@pmu.ac.at.
Heidemarie DobiasLaboratory for Tumour Biology and Experimental Therapies (TREAT), Institute of Physiology and Pathophysiology, Paracelsus Medical University Salzburg, Strubergasse 22, 5020 Salzburg, Austria. heidi.dobias@pmu.ac.at.
Martin GaisbergerDepartment for Radon Therapy Research, Ludwig Boltzmann Cluster for Arthritis and Rehabilitation, Institute of Physiology and Pathophysiology, Paracelsus Medical University Salzburg, Strubergasse 22, 5020 Salzburg, Austria. martin.gaisberger@pmu.ac.at.
Martin PichlerDivision of Oncology, Department of Internal Medicine, Medical University Graz, Auenbruggerplatz 15, 8036 Graz, Austria. martin.pichler@medunigraz.at.
Markus RitterLaboratory for Tumour Biology and Experimental Therapies (TREAT), Institute of Physiology and Pathophysiology, Paracelsus Medical University Salzburg, Strubergasse 22, 5020 Salzburg, Austria. markus.ritter@pmu.ac.at.
Martin JakabLaboratory of Functional and Molecular Membrane Physiology (FMMP), Institute of Physiology and Pathophysiology, Paracelsus Medical University Salzburg, Strubergasse 22, 5020 Salzburg, Austria. martin.jakab@pmu.ac.at.
Daniel NeureiterInstitute of Pathology, Paracelsus Medical University/Salzburger Landeskliniken (SALK), Muellner Hauptstrasse 48, 5020 Salzburg, Austria. d.neureiter@salk.at.
Tobias KiesslichLaboratory for Tumour Biology and Experimental Therapies (TREAT), Institute of Physiology and Pathophysiology, Paracelsus Medical University Salzburg, Strubergasse 22, 5020 Salzburg, Austria. t.kiesslich@salk.at.
Paracelsus Medical University · ATThe University of Texas MD Anderson Cancer Center · US

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

The clonogenic assay is a widely used method to study the ability of cells to 'infinitely' produce progeny and is, therefore, used as a tool in tumor biology to measure tumor-initiating capacity and stem cell status. However, the standard protocol of using 6-well plates has several disadvantages. By miniaturizing the assay to a 96-well microplate format, as well as by utilizing the confluence detection function of a multimode reader, we here describe a new and modified protocol that allows comprehensive experimental setups and a non-endpoint, label-free semi-automatic analysis. Comparison of bright field images with confluence images demonstrated robust and reproducible detection of clones by the confluence detection function. Moreover, time-resolved non-endpoint confluence measurement of the same well showed that semi-automatic analysis was suitable for determining the mean size and colony number. By treating cells with an inhibitor of clonogenic growth (PTC-209), we show that our modified protocol is suitable for comprehensive (broad concentration range, addition of technical replicates) concentration- and time-resolved analysis of the effect of substances or treatments on clonogenic growth. In summary, this protocol represents a time- and cost-effective alternative to the commonly used 6-well protocol (with endpoint staining) and also provides additional information about the kinetics of clonogenic growth.

Indexed as

Cell Line, TumorCytostatic AgentsHeterocyclic Compounds, 2-RingHumansMiniaturizationThiazolesTumor Stem Cell AssayCytostatic AgentsHeterocyclic Compounds, 2-RingPTC-209Thiazoles96-well microplate formatclonogenic assayclonogenic growthconfluence detectionmicroplate reader

Identifiers

PMID29510509
PMCPMC5877585
OpenAlexW2794209007

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.