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RNA-seq Analysis of Transcriptomes in Thrombin-treated and Control Human Pulmonary Microvascular Endothelial Cells

RNA-seq Analysis of Transcriptomes in Thrombin-treated and Control Human Pulmonary Microvascular Endothelial Cells

Published on: February 13, 2013

Transcript profiling of human platelets using microarray and serial analysis of gene expression (SAGE).

Dmitri V Gnatenko¹, John J Dunn, John Schwedes

¹Division of Hematology/Oncology, Department of Medicine, State University of New York at Stony Brook, Stony Brook, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)

|October 8, 2008

Summary

This summary is machine-generated.

Platelets, lacking DNA, possess a unique transcriptome of 1,600-3,000 mRNAs. This study details methods for platelet purification and transcript profiling using microarray and SAGE techniques.

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Isolation and Profiling of Human Primary Mesenteric Arterial Endothelial Cells at the Transcriptome Level

Isolation and Profiling of Human Primary Mesenteric Arterial Endothelial Cells at the Transcriptome Level

Published on: March 14, 2022

Area of Science:

Hematology
Molecular Biology
Genomics

Background:

Platelets are anucleated cells derived from megakaryocytes through thrombopoiesis.
Despite lacking genomic DNA, platelets contain a substantial pool of mRNA transcripts, forming a platelet transcriptome.
This platelet transcriptome exhibits a unique and reproducible profile, comprising approximately 1,600-3,000 distinct mRNA transcripts.

Purpose of the Study:

To provide detailed protocols for platelet purification.
To describe methods for transcript profiling in platelets using specific platforms.
To discuss the unique considerations in platelet purification, RNA isolation, and transcript analysis.

Main Methods:

Detailed protocols for large-scale platelet purification from apheresis bags (3-5 x 10^11 platelets/bag).
Modified small-scale platelet purification protocols for isolating platelets from 20 ml of peripheral blood.
Transcript profiling using Affymetrix microarray and Serial Analysis of Gene Expression (SAGE) platforms.

Main Results:

Established reproducible protocols for purifying platelets from various blood volumes.
Successfully applied microarray and SAGE for comprehensive platelet transcriptome analysis.
Identified and characterized the unique mRNA profile within platelets.

Conclusions:

Platelet purification and transcript profiling are feasible using the described methodologies.
The characterized platelet transcriptome provides a basis for understanding platelet function and biology.
Detailed protocols facilitate further research into platelet mRNA content and its implications.