Assessing the Impact of Cell Isolation Methods on Gene Expression in CD19+ B cells Using Next Generation Sequencing

Authors

• D Owusu-Ansah
• A Henning
• V Degiorgi

Abstract

The targeted isolation of B-cell cell populations is crucial for researchers investigating lymphoproliferative disorders, particularly when investigating non-Hodgkin’s lymphoma. Over the years, cell isolation methods have evolved significantly; however, the potential influence of these methods on downstream gene expression in target cell populations remains a subject of interest. It is widely believed that techniques such as positive magnetic activated cell sorting (MACS) and fluorescence-activated cell sorting (FACS) may affect the expression of genes due to the direct interaction of associated antibodies with surface markers.
We currently know through literature review that cofactors like pressure, nozzle size, instrument type, have little effect on gene expression profile. We also know that temperature can play a significant role on downstream expression. However, to the best of our knowledge, no specific studies have examined these effects within CD19+ B cell populations. In this study, we plan to isolate B cells from n=4 healthy donors using three distinct methods: positive MACS, negative MACS, and FACS. We will then perform Next Generation Sequencing (NGS) analysis in order to identify differentially expressed genes among the three different isolation methods.
In this study we will be comparing the performance of three isolation methods and their effect on gene expression in CD19+ B cells, trying to establish if different cell isolation methods would potentially impact the disease-related signature when analyzing patient samples.

Scientific Focus Area: Immunology

This page was last updated on Monday, September 25, 2023