Preparation of Myeloid Derived Suppressor Cells (MDSC) from Naive and Pancreatic Tumor-bearing Mice using Flow Cytometry and Automated Magnetic Activated Cell Sorting (AutoMACS)

Overview

This article presents a rapid method for immunophenotyping Myeloid Derived Suppressor Cells (MDSC) and enriching Gr-1 + leukocytes from mouse spleens. The technique employs flow cytometry and AutoMACS Cell Sorting to isolate viable Gr-1 + leukocytes for subsequent assays.

Key Study Components

Area of Science

• Immunology
• Cell Biology
• Flow Cytometry

Background

• Myeloid Derived Suppressor Cells (MDSC) play a crucial role in immune regulation.
• Enrichment of specific leukocyte populations is essential for various in vivo and in vitro studies.
• Flow cytometry is a powerful tool for analyzing cell populations.
• AutoMACS Cell Sorting enhances the efficiency of cell isolation.

Purpose of Study

• To develop a rapid protocol for enriching Gr-1 + leukocytes from mouse spleens.
• To facilitate the study of MDSC in different experimental contexts.
• To improve the viability and yield of isolated cells for assays.

Methods Used

• Pooled mouse spleens are processed to create a single cell suspension.
• Flow cytometric analysis is performed to quantify MDSC prior to enrichment.
• Cells are labeled with antibodies against Gr-1 and processed with magnetic microbeads.
• AutoMACS pro separator is used to separate Gr-1 positive and negative populations.

Main Results

• The method successfully enriches Gr-1 + leukocytes from mouse spleens.
• High viability of isolated MDSC is achieved for further assays.
• Flow cytometry confirms the effectiveness of the enrichment process.
• The protocol can be adapted for various experimental needs.

Conclusions

• This rapid method enhances the study of MDSC and their functions.
• It provides a reliable approach for isolating specific leukocyte populations.
• The technique is beneficial for both in vivo and in vitro research applications.

Frequently Asked Questions