Isolation of Myeloid Dendritic Cells and Epithelial Cells from Human Thymus

Christina Stoeckle; Ioanna A. Rota; Eva Tolosa; Christoph Haller; Arthur Melms; Eleni Adamopoulou

doi:10.3791/50951

JoVE Journal
Immunology and Infection

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Immunology and Infection

Isolation of Myeloid Dendritic Cells and Epithelial Cells from Human Thymus

인간의 흉선에서 골수 수지상 세포와 상피 세포의 분리

Full Text

20,031 Views

09:07 min

September 19, 2013

DOI: 10.3791/50951-v

Christina Stoeckle^1,2, Ioanna A. Rota¹, Eva Tolosa³, Christoph Haller⁴, Arthur Melms⁵, Eleni Adamopoulou¹

¹Department of General Neurology,Hertie Institute for Clinical Brain Research, ²Institute of Pharmacology,University of Bern, ³Department of Immunology,University Medical Center Hamburg-Eppendorf, ⁴Department of Thoracic and Cardiovascular Surgery,University Clinic Tuebingen, ⁵Department of Neurology,University Hospital Erlangen

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol outlines a method for isolating dendritic and epithelial cells from the human thymus. The process involves mechanical disruption, enzymatic digestion, and density centrifugation to obtain a single cell suspension.

Key Study Components

Area of Science

Cell Biology
Immunology
Neuroscience

Background

Dendritic cells and thymic epithelial cells are crucial for immune function.
Isolating these cells can enhance our understanding of immune responses.
Traditional methods may not efficiently isolate these rare cell populations.
This protocol aims to provide a reproducible and cost-effective approach.

Purpose of Study

To isolate dendritic and epithelial cells from human thymus tissue.
To improve methods for studying rare cell populations.
To facilitate research in immunology and related fields.

Methods Used

Mechanical disruption of thymus tissue.
Enzymatic digestion to create a single cell suspension.
Percoll density centrifugation for cell enrichment.
Magnetic or FACS sorting for cell isolation.

Main Results

Successful isolation of dendritic and epithelial cells.
High purity of isolated cell populations.
Reproducibility of the method across different samples.
Cost-effectiveness compared to traditional methods.

Conclusions

The protocol provides an efficient method for isolating rare cells.
It can be applied in various research settings.
Future studies can leverage this method for deeper insights into immune function.

Frequently Asked Questions

What types of cells can be isolated using this protocol?

This protocol allows for the isolation of dendritic cells and epithelial cells from the human thymus.

Is this method cost-effective?

Yes, the method is designed to be reproducible and cost-effective compared to traditional isolation techniques.

What is the first step in the isolation process?

The first step involves mechanically disrupting the dissected thymus tissue.

How are the cells enriched after digestion?

Cells are enriched using Percoll density centrifugation to separate low-density fractions.

Can this method be used for other types of tissues?

While this protocol is specific to the human thymus, similar methods may be adapted for other tissues.

What sorting techniques are used for cell isolation?

Cells can be isolated using magnetic sorting or FACS sorting, or a combination of both.

이 프로토콜은 항원 밀도 단일 세포 현탁액을 원심 분리하고 그 세포 집단의 마지막 자석 및 / 또는 FACS 정렬 뒤에 조직의 소화 효소의 여러 단계를 통해 인간 흉선에서 제시 세포 분리하는 방법을 자세히 설명.

이 절차의 전반적인 목표는 인간 흉선에서 수지상 세포와 상피 세포를 분리하는 것입니다. 이것은 먼저 기계적 파괴에 의해 절개된 조직을 단편화함으로써 달성됩니다. 다음으로, 조직 단편을 추가로 효소로 소화하여 단일 세포 현탁액을 생성합니다.

마지막으로, 세포 현탁액은 perca 분리에 의해 세포의 저밀도 분획에 대해 농축됩니다. 궁극적으로, 수지상 세포와 상피 세포는 자기 또는 사실 분류, 또는 둘의 조합에 의해 이 분획으로부터 분리될 수 있습니다. 여기에서 제시하는 방법은 인간 흉선에서 발견되는 수지상 세포 및 흉선 상피 세포와 같은 희귀 세포 집단의 성공적인 분리 및 후속 연구를 위한 효율적이고 재현 가능하며 비용 효율적인 절차를 제공합니다.

View the full transcript and gain access to thousands of scientific videos