Oral mucosa dissociation protocols for targeting fibroblasts

Authors

• I Pavlinov
• Q Zhang
• M Kosikova
• HN Nguyen
• H Xie
• W Zheng

Abstract

Periodontitis is one of the most common diseases associated with the oral cavity resulting in mucosal inflammation and bone loss. It has been shown that immune cells such as neutrophils play pathogenic roles in periodontitis, however, we do not know the role of non-immune cells. Our lab recently published scRNAseq data in humans showing that stromal cells may recruit neutrophils to the oral mucosa leading to inflammation. Oral fibroblasts play an important role in wound healing and tissue repair in the gingival connective tissue. During further studies of oral fibroblasts in mice, we discovered that the enzyme selection for tissue dissociation influences the amount of fibroblasts recovered. In this current study, we compared three different tissue dissociation methods: collagenase ‚Ö£, collagenase ‚Ö° and enzymes in Miltenyi Whole skin dissociation kit in the process of oral mucosal tissue dissociation to obtain fibroblasts as single cells. We evaluated fibroblasts using flow cytometry by cell surface markers. There were significant differences in the viabilities and fibroblasts proportions between the methods. We conclude using collagenase ‚Ö° is the best dissociation method to obtain high viability and amount of fibroblasts from oral mucosa.

Scientific Focus Area: Immunology

This page was last updated on Monday, September 25, 2023