Anaerobic Co-Culture of Mesenchymal Stem Cells and Anaerobic Pathogens - A New In Vitro Model System
Authors
Katja Kriebel, Anne Biedermann, Bernd Kreikemeyer, Hermann Lang
Institution
University Rostock
Country
Germany
Year
2013
Journal
PLOS
Abstract
Background: Human mesenchymal stem cells (hMSCs) are multipotent by nature and are originally isolated from bone
marrow. In light of a future application of hMSCs in the oral cavity, a body compartment with varying oxygen partial
pressures and an omnipresence of different bacterial species i.e. periodontitis pathogens, we performed this study to gain
information about the behavior of hMSC in an anaerobic system and the response in interaction with oral bacterial
pathogens.
Methodology/Principal Findings: We established a model system with oral pathogenic bacterial species and eukaryotic
cells cultured in anaerobic conditions. The facultative anaerobe bacteria Fusobacterium nucleatum, Porphyromonas gingivalis
and Aggregatibacter actinomycetemcomitans were studied. Their effects on hMSCs and primary as well as permanent
gingival epithelial cells (Ca9-22, HGPEC) were comparatively analyzed. We show that hMSCs cope with anoxic conditions,
since 40% vital cells remain after 72 h of anaerobic culture. The Ca9-22 and HGPEC cells are significantly more sensitive to
lack of oxygen. All bacterial species reveal a comparatively low adherence to and internalization into hMSCs (0.2% and
0.01% of the initial inoculum, respectively). In comparison, the Ca9-22 and HGPEC cells present better targets for bacterial
adherence and internalization. The production of the pro-inflammatory chemokine IL-8 is higher in both gingival epithelial
cell lines compared to hMSCs and Fusobacterium nucleatum induce a time-dependent cytokine secretion in both cell lines.
Porphyromonas gingivalis is less effective in stimulating secretion of IL-8 in the co-cultivation experiments.
Conclusions/significance: HMSCs are suitable for use in anoxic regions of the oral cavity. The interaction with local
pathogenic bacteria does not result in massive pro-inflammatory cytokine responses. The test system established in this
study allowed further investigation of parameters prior to set up of oral hMSC in vivo studies