Autoren:	Geiß, Carsten Alanis-Lobato, Gregorio Andrade, Miguel Régnier-Vigouroux, Anne
Titel:	Assessing the reliability of gene expression measurements in very-low-numbers of human monocyte-derived macrophages
Online-Publikationsdatum:	20-Dez-2019
Erscheinungsdatum:	2019
Sprache des Dokuments:	Englisch
Zusammenfassung/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-Sachgruppe:	570 Biowissenschaften 570 Life sciences
Veröffentlichende Institution:	Johannes Gutenberg-Universität Mainz
Organisationseinheit:	FB 10 Biologie
Veröffentlichungsort:	Mainz
ROR:	https://ror.org/023b0x485
DOI:	http://doi.org/10.25358/openscience-245
URN:	urn:nbn:de:hebis:77-publ-594849
Version:	Published version
Publikationstyp:	Zeitschriftenaufsatz
Nutzungsrechte:	CC BY
Informationen zu den Nutzungsrechten:	https://creativecommons.org/licenses/by/4.0/
Zeitschrift:	Scientific reports 9
Seitenzahl oder Artikelnummer:	Art. 17908
Verlag:	Macmillan Publishers Limited, part of Springer Nature
Verlagsort:	London
Erscheinungsdatum:	2019
ISSN:	2045-2322
URL der Originalveröffentlichung:	http://dx.doi.org/10.1038/s41598-019-54500-8
DOI der Originalveröffentlichung:	10.1038/s41598-019-54500-8
Anmerkung:	Andrade, Miguel veröffentlicht unter: Andrade-Navarro, Miguel
Enthalten in den Sammlungen:	JGU-Publikationen