בידוד ושימור בהקפאה של תאים חד-גרעיניים אנושיים היקפיים בעלי יכולת קיימא גבוהה מדם שלם: מדריך למתחילים

Overview

This protocol provides a step-by-step guide for collecting, storing, and thawing peripheral blood mononuclear cells (PBMCs) from whole blood. It also includes methods for collecting plasma and buffy coat, which are essential for various downstream analyses.

Key Study Components

Area of Science

• Cell biology
• Immunology
• Hematology

Background

• Peripheral blood mononuclear cells (PBMCs) include lymphocytes, dendritic cells, and monocytes.
• PBMCs are widely used in research for various assays.
• High viability PBMCs are crucial for accurate experimental results.
• Collection methods impact the quality and quantity of cells obtained.

Purpose of Study

• To provide a reliable protocol for PBMC collection and storage.
• To ensure high viability of cells for downstream applications.
• To demonstrate the collection of additional components like plasma and buffy coat.

Methods Used

• Collection of PBMCs from whole blood using density gradient centrifugation.
• Preparation of materials and reagents as outlined in the protocol.
• Storage and thawing procedures for PBMCs.
• Collection techniques for EDTA plasma and buffy coat.

Main Results

• Successful collection of approximately 12 million PBMCs per tube.
• High viability rates of PBMCs suitable for downstream analyses.
• Demonstrated methods for plasma and buffy coat collection.
• Protocol facilitates various experimental workflows including RNA sequencing and flow cytometry.

Conclusions

• This protocol provides a comprehensive guide for PBMC collection.
• Ensures high-quality samples for immunological and cellular studies.
• Facilitates further research in various biological assays.

Frequently Asked Questions