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http://doi.org/10.25358/openscience-245| 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 |
| ROR: | https://ror.org/023b0x485 |
| DOI: | http://doi.org/10.25358/openscience-245 |
| URN: | urn:nbn:de:hebis:77-publ-594849 |
| Version: | Published version |
| Publication type: | Zeitschriftenaufsatz |
| License: | CC BY |
| Information on rights of use: | https://creativecommons.org/licenses/by/4.0/ |
| Journal: | Scientific reports 9 |
| 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: | http://dx.doi.org/10.1038/s41598-019-54500-8 |
| Publisher DOI: | 10.1038/s41598-019-54500-8 |
| Annotation: | Andrade, Miguel veröffentlicht unter: Andrade-Navarro, Miguel |
| Appears in collections: | JGU-Publikationen |
