Purification and Expansion of Mouse Invariant Natural Killer T Cells for in vitro and in vivo Studies

Overview

This protocol enables the purification and expansion of invariant natural killer T (iNKT) cells from mouse spleen. The process allows for a 30-fold increase in cell numbers, making them suitable for both in vitro and in vivo studies.

Key Study Components

Area of Science

• Immunology
• Cell Biology
• Neuroscience

Background

• iNKT cells are rare immune cells with significant roles in immune responses.
• Purification of these cells is challenging due to their low abundance.
• Expanding iNKT cells is crucial for various research applications.
• This protocol aims to streamline the process of obtaining and expanding these cells.

Purpose of Study

• To provide a rapid and efficient method for iNKT cell purification.
• To enable significant expansion of iNKT cells for research purposes.
• To facilitate the use of iNKT cells in both in vitro and in vivo studies.

Methods Used

• Dissection of mouse spleen to obtain a single cell suspension.
• Use of a 70 nanometer cell strainer for cell purification.
• Centrifugation to isolate the cell pellet.
• Resuspension of the pellet in ammonium chloride potassium lysine buffer.

Main Results

• Successful purification of iNKT cells in one day.
• Achieved a 30-fold expansion of iNKT cells within two weeks.
• Produced a large number of ready-to-use iNKT cells for experiments.
• Demonstrated by Gloria Delfanti, a post-doc in the lab.

Conclusions

• The protocol provides a reliable method for iNKT cell enrichment.
• It significantly increases the availability of these cells for research.
• This approach can enhance studies involving immune responses.

Frequently Asked Questions