Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

0

NanoString- Transcriptional profiling of LPS-primed and inflammasome-activated untouched, or platelet-depleted human CD14+ monocytes


ABSTRACT: The goal of this experiment was to explore how human platelets affect the transcriptional responses of primary human CD14+ blood monocytes to lipopolysaccharide (LPS), and NLRP3 activation with Nigericin. For this purpose, we analyzed the mRNA expression of 770 myeloid-specific transcripts using the nCounter® Nanostring Human Myeloid Innate Immunity Panel v2. We isolated classical monocytes (CD14+CD16−) from the peripheral blood of healthy volunteers. Monocytes were derived from PBMCs using negative selection with the EasySep™ Human Monocyte Isolation Kit from STEMCELLTM Technologies. We modified this process by either including or excluding a platelet-depleting cocktail, creating two groups: \\"Standard Monocytes (StdMo)\\" and \\"Platelet-depleted Monocytes (PdMo).\\" To examine the impact of platelets further, we supplemented PdMos with fresh autologous platelets at a ratio of 50 platelets per monocyte, resulting in a third group, \\"PdMo + Plts.\\" StdMos were prepared according to the standard protocol provided by the EasySep™ Kit. For the PdMos, a Platelet Removal Component (50 µl ml−1) from the kit was used during isolation. We also reconstituted platelet-depleted monocytes with platelets and analyzed platelets alone to determine their specific mRNA contributions. The groups—StdMo, PdMo, PdMo + Plts, and platelets alone—were then exposed to 2 ng/ml of Ultrapure LPS (from E. coli O111:B4) for 4.5 hours, or stimulated with LPS for 3 hours followed by inflammasome activation with Nigericin (10 µM) for 90 min before extracting RNA for analysis.

ORGANISM(S): Homo sapiens

SUBMITTER: Bernardo Franklin S 

PROVIDER: E-MTAB-14072 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

Similar Datasets

2015-06-22 | E-GEOD-59421 | biostudies-arrayexpress
2008-06-18 | E-GEOD-11524 | biostudies-arrayexpress
2021-12-08 | PXD026903 | Pride
2016-10-10 | PXD003365 | Pride
2016-10-10 | PXD003363 | Pride
2015-04-27 | E-MTAB-2355 | biostudies-arrayexpress
2014-02-04 | E-GEOD-51453 | biostudies-arrayexpress
2024-02-15 | PXD045535 | Pride
2014-07-17 | E-GEOD-59488 | biostudies-arrayexpress
2014-01-01 | E-GEOD-45960 | biostudies-arrayexpress