Cryopreservation and Bioenergetic Evaluation of Human Peripheral Blood Mononuclear Cells

Fabian Dieter; Jendrik Grube; Tim Birkenhauer; Alice Quentin; Gunter P. Eckert

doi:10.3791/65730

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Immunology and Infection
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JoVE Journal Immunology and Infection

Cryopreservation and Bioenergetic Evaluation of Human Peripheral Blood Mononuclear Cells

인간 말초 혈액 단핵 세포의 냉동 보존 및 생체 에너지 평가

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04:10 min

October 20, 2023

DOI: 10.3791/65730-v

Fabian Dieter¹, Jendrik Grube¹, Tim Birkenhauer¹, Alice Quentin¹, Gunter P. Eckert¹

¹Biomedical Research Center, Institute of Nutritional Sciences,Justus-Liebig-University of Giessen

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Overview

This article presents a standardized method for the preparation and cryopreservation of peripheral blood mononuclear cells (PBMCs). The protocol aims to maintain bioenergetic parameters post-thawing, ensuring reliable measurements in immune function studies.

Key Study Components

Area of Science

Immunology
Cell Biology
Bioenergetics

Background

PBMCs are crucial for analyzing immune functions and disorders.
Cryopreservation allows for time-independent sample analysis.
Freezing and thawing can damage cells, affecting experimental outcomes.
Standardized methods enhance reproducibility across studies.

Purpose of Study

To develop a reliable cryopreservation protocol for PBMCs.
To ensure bioenergetic parameters remain unchanged after thawing.
To facilitate multi-center studies by preserving samples for later analysis.

Methods Used

Standardized preparation of PBMCs from whole blood.
Controlled cryopreservation techniques.
Assessment of bioenergetic parameters post-thawing.
Comparison of results across different sample collection sites.

Main Results

The protocol successfully preserves PBMCs without damaging them.
Bioenergetic measurements remain consistent after thawing.
Samples can be collected from various locations and analyzed together.

Conclusions

Reliable cryopreservation of PBMCs enhances research capabilities.
Standardized methods improve the quality of immune function studies.
This approach supports collaborative research across multiple sites.

Frequently Asked Questions

What are PBMCs?

PBMCs, or peripheral blood mononuclear cells, are a type of blood cell that plays a crucial role in the immune system.

Why is cryopreservation important?

Cryopreservation allows for the long-term storage of cells, enabling time-independent analysis and multi-center studies.

What challenges are associated with cryopreservation?

The freezing and thawing process can damage cells, affecting their viability and the accuracy of subsequent measurements.

How does this protocol improve PBMC preservation?

The protocol minimizes cell damage during freezing and thawing, ensuring that bioenergetic parameters are preserved.

Can samples be analyzed at different locations?

Yes, the protocol allows for samples to be collected from various centers and analyzed at a single location.

What types of studies can benefit from this method?

Studies focusing on immune functions, metabolic diseases, and mitochondrial functions can benefit from this cryopreservation method.

분리된 말초 혈액 단핵 세포는 면역 기능 및 장애, 대사 질환 또는 미토콘드리아 기능의 분석에 사용할 수 있습니다. 이 연구에서는 전혈에서 PBMC를 준비하고 후속 냉동 보존을 위한 표준화된 방법을 설명합니다. 냉동 보존은 이 시간과 장소를 독립적으로 만듭니다.

이 방법의 도움으로 해동 후 생체 에너지 매개변수가 변경되지 않는 방식으로 PBMC를 동결 보존하는 데 적합한 절차를 설명하고자 합니다. 냉동 보존의 주요 실험 과제 중 하나는 냉동 및 해동 과정이 PBMC를 손상시킬 수 있으며, 이는 다음과 같은 생체 에너지 측정에 영향을 미친다는 것입니다. 당사의 프로토콜을 통해 측정된 생체 에너지 매개변수가 영향을 받지 않도록 세포에 손상을 주지 않고 PBMC를 안정적으로 보존하는 방법을 보여주려고 합니다.

PBMC를 보존할 수 있기 때문에 여러 센터에서 샘플을 수집하고 한 위치에서 측정할 수 있습니다. 또한 측정 시간을 더 독립적으로 만듭니다. PBMC의 냉동 보존을 통해 세포를 수집하고 보존하는 여러 부위에서 동시에 연구를 수행할 수 있습니다.

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