Isolation of myeloid dendritic cells and epithelial cells from human thymus

Stoeckle, Christina; Rota, Ioanna A.; Tolosa, Eva; Haller, Christoph; Melms, Arthur; Adamopoulou, Eleni (2013). Isolation of myeloid dendritic cells and epithelial cells from human thymus. Journal of visualized experiments, 79(79), e50951. MYJoVE Corporation 10.3791/50951

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In this protocol we provide a method to isolate dendritic cells (DC) and epithelial cells (TEC) from the human thymus. DC and TEC are the major antigen presenting cell (APC) types found in a normal thymus and it is well established that they play distinct roles during thymic selection. These cells are localized in distinct microenvironments in the thymus and each APC type makes up only a minor population of cells. To further understand the biology of these cell types, characterization of these cell populations is highly desirable but due to their low frequency, isolation of any of these cell types requires an efficient and reproducible procedure. This protocol details a method to obtain cells suitable for characterization of diverse cellular properties. Thymic tissue is mechanically disrupted and after different steps of enzymatic digestion, the resulting cell suspension is enriched using a Percoll density centrifugation step. For isolation of myeloid DC (CD11c(+)), cells from the low-density fraction (LDF) are immunoselected by magnetic cell sorting. Enrichment of TEC populations (mTEC, cTEC) is achieved by depletion of hematopoietic (CD45(hi)) cells from the low-density Percoll cell fraction allowing their subsequent isolation via fluorescence activated cell sorting (FACS) using specific cell markers. The isolated cells can be used for different downstream applications.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Pharmacology

UniBE Contributor:

Merz, Christina

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1940-087X

Publisher:

MYJoVE Corporation

Language:

English

Submitter:

Anita Dähler

Date Deposited:

30 Jan 2014 16:35

Last Modified:

15 Dec 2017 09:44

Publisher DOI:

10.3791/50951

PubMed ID:

24084687

Uncontrolled Keywords:

Immunology, Issue 79, Immune System Processes, Biological Processes, immunology, Immune System Diseases, Immune System Phenomena, Life Sciences (General), immunology, human thymus, isolation, dendritic cells, mTEC, cTEC

BORIS DOI:

10.7892/boris.39628

URI:

https://boris.unibe.ch/id/eprint/39628

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