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Laryngorhinootologie 2013; 92(07): 462-469
DOI: 10.1055/s-0033-1337982
Originalie
© Georg Thieme Verlag KG Stuttgart · New York

Genexpressionsmuster mesenchymaler Stammzellen: ein ungelöster Aspekt in der regenerativen Medizin

Trying to Unravel an Unresolved Issue in Regenerative Medicine: Gene Expression Profiling of MSCs
M. Jakob
1   University Hospital Bonn, ENT, Bonn
,
K. Bruderek
2   University Hospital Essen, ENT, Essen
,
F. Bootz
1   University Hospital Bonn, ENT, Bonn
,
S. Lang
2   University Hospital Essen, ENT, Essen
,
S. Brandau
2   University Hospital Essen, ENT, Essen
› Institutsangaben
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Publikationsverlauf

eingereicht25. September 2012

akzeptiert 20. Februar 2013

Publikationsdatum:
16. April 2013 (online)

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Zusammenfassung

Hintergrund:

Aufgrund der antiinflammatorischen und regenerativen Wirkung spielen mesen­chymale Stammzellen (MSCs) bei der Gewebe­regeneration, der Inflammation und bei der Gewebeschädigung eine wichtige Rolle. Gewebeschädigung, wie z. B. bei Bestrahlung oder Infektion, setzt sogenannte Gefahrenstoffe von sterbenden Zellen oder eindringenden patho­genen Keimen im Gewebe frei. Die molekulare Antwort von MSCs in diesem Prozess des Gewebe­stresses bleibt derzeit weitestgehend ­unklar.

Material und Methoden:

MSCs der Glandula Parotidea (pgMSCs) von 3 Spendern wurden isoliert, in Zellkultur vermehrt und in die 3 klassischen plus der myogenen Differenzierungslinien differenziert. Wir haben die zellbiologische Antwort dieser pgMSCs anhand eines Gewebestressmodells durch Stimulation mit dem bakteriellen Endotoxin Lipopolysaccharid (LPS) in vitro ­geprüft. Ausgewertet wurde das Genexpressionsmuster der MSCs mittels eines Gen-Arrays.

Ergebnisse:

Durch Immunfluoreszenz und Immunhistochemie konnten wir stammzelltypisches osteogenes, adipogenes, chondrogenes und myogenes Differenzierungspotenzial nachweisen. PgMSCs zeigten nach LPS Stimulation auf Genexpressionsebene eine signifikante Aktivierung immunologisch wichtiger Signalwege. So werden typische an der Immunregulation beteiligte Rezeptoren und Liganden der MSCs, wie Interleukine, TGF-β, Tumornekrosefaktoren (TNF) und Toll-ähnliche Rezeptoren (TLR), reguliert.

Schlussfolgerung:

Die vorliegende Studie entschlüsselt wichtige Aspekte der molekularen Antwort von gewebeständigen MSCs bei Inflammation und Gewebeschädigung. Das Verständnis dieser zellbiologischen Funktionen ist Vorraussetzung für den zukünftigen Einsatz von MSCs als Therapeutikum.

Abstract

Trying to Unravel an Unresolved Issue in Regenera­tive Medicine: Gene Expression Profiling of MSCs

Background:

Mesenchymal stem cells (MSCs) are adult fibroblastoid progenitor cells. Because of their immunoregulatory properties and their so-called trophic effects, MSCs play an important role in tissue regeneration, inflammation and trauma. Tissue trauma and challenge, for example during radiotherapy or infection, result in the release of so-called “danger molecules”, which may be derived from dying cells or incoming pathogens. The molecular response of MSCs to this tissue stress remains largely elusive.

Material and Methods:

In this study we examined the cell biological response of MSCs derived from human parotid glands (pgMSCs) and used bacterial endotoxin as a model of tissue stress and inflammation. PgMSCs from 3 donors were isolated, expanded and tested for classical tri-lineage plus myogenic differentiation. The cell biological response to the model “stressor” endotoxin was examined by low density gene expression arrays.

Results:

Through immunofluorescence and immunohistochemistry we were able to proof osteogenic, adipogenic, chondrogenic, and myogenic differentiation potential characteristic for stem cells. In vitro, gene expression analysis showed a characteristic modulation of MSCs after stimulation with endotoxin Lipopolysaccharide (LPS). Specifically, receptors and ligands typically involved in immune regulation, such as interleukins, TGF-β, tumor necrosis factors (TNF), and toll-like receptors (TLR), were regulated.

Conclusion:

Our study elucidates some key functions and molecules, which are regulated in MSCs during tissue stress and inflammation. A thorough understanding of their cell biological function will aid future rationale therapeutic application of MSCs.

Schlüsselwörter

mesenchymale Stammzellen - Ohrspeicheldrüse - Genexpressionsmuster - Differenzierung - Inflammation

Key words

mesenchymal stem cells - salivary glands - gene expression profiling - differentiation - inflammation
 

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