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Authors: Geiß, Carsten
Alanis-Lobato, Gregorio
Andrade, Miguel
Régnier-Vigouroux, Anne
Title: Assessing the reliability of gene expression measurements in very-low-numbers of human monocyte-derived macrophages
Online publication date: 20-Dec-2019
Year of first publication: 2019
Language: english
Abstract: Tumor-derived primary cells are essential for in vitro and in vivo studies of tumor biology. The scarcity of this cellular material limits the feasibility of experiments or analyses and hence hinders basic and clinical research progress. We set out to determine the minimum number of cells that can be analyzed with standard laboratory equipment and that leads to reliable results, unbiased by cell number. A proof-of-principle study was conducted with primary human monocyte-derived macrophages, seeded in decreasing number and constant cell density. Gene expression of cells stimulated to acquire opposite inflammatory states was analyzed by quantitative PCR. Statistical analysis indicated the lack of significant difference in the expression profile of cells cultured at the highest (100,000 cells) and lowest numbers (3,610 cells) tested. Gene Ontology, pathway enrichment and network analysis confirmed the reliability of the data obtained with the lowest cell number. This statistical and computational analysis of gene expression profiles indicates that low cell number analysis is as dependable and informative as the analysis of a larger cell number. Our work demonstrates that it is possible to employ samples with a scarce number of cells in experimental studies and encourages the application of this approach on other cell types.
DDC: 570 Biowissenschaften
570 Life sciences
Institution: Johannes Gutenberg-Universität Mainz
Department: FB 10 Biologie
Place: Mainz
URN: urn:nbn:de:hebis:77-publ-594849
Version: Published version
Publication type: Zeitschriftenaufsatz
License: CC BY
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Journal: Scientific reports
Pages or article number: Art. 17908
Publisher: Macmillan Publishers Limited, part of Springer Nature
Publisher place: London
Issue date: 2019
ISSN: 2045-2322
Publisher URL:
Publisher DOI: 10.1038/s41598-019-54500-8
Annotation: Andrade, Miguel veröffentlicht unter: Andrade-Navarro, Miguel
Appears in collections:JGU-Publikationen

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