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Isolation and Cryopreservation of Highly Viable Human Peripheral Blood Mononuclear Cells From Whole Blood: A Guide for Beginners

2.3K Views

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10:48 min

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October 25th, 2024

DOI :

10.3791/66794-v

October 25th, 2024

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Brian Dinh¹, Marten A. Hoeksema², Nathanael J. Spann³, Jacob Rendler¹, Isidoro Cobo⁴^,⁵, Christopher K. Glass³^,⁶, Calvin Yeang¹

¹Division of Cardiology, Department of Medicine, University of California San Diego, ²Department of Medical Biochemistry, Amsterdam Immunity and Infection: Inflammatory diseases; Amsterdam Cardiovascular Sciences: Atherosclerosis and Ischemic Syndrome, Amsterdam UMC location University of Amsterdam, ³Department of Cellular and Molecular Medicine, University of California San Diego, ⁴Division of Clinical Immunology & Rheumatology, Department of Medicine, Heersink School of Medicine, University of Alabama at Birmingham, ⁵Comprehensive Arthritis, Musculoskeletal, Bone and Autoimmunity Center, Heersink School of Medicine, University of Alabama at Birmingham, ⁶Department of Medicine, University of California San Diego

Summary

This protocol presents an accessible guide for collecting, storing, and thawing peripheral blood mononuclear cells suitable for downstream analyses and workflows like flow cytometry and RNA sequencing. Plasma and buffy coat collections are also demonstrated.

Explore More Videos

Cryopreservation

Human Peripheral Blood Mononuclear Cells

Dendritic Cells

Density Gradient Centrifugation

Whole Buffy Coat

Phlebotomy Technique

Centrifugation Protocol

Assay Applications

Chapters in this video

0:08

Introduction

1:02

Processing & Cryopreservation of PBMCs

5:12

Processing of Plasma

6:17

Thawing PBMCs

8:15

Representative Results

10:13

Discussion

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