Definition of Different Cell Populations in Heterogeneous Primary DFSC Cultures

Objectives: The goal of the research is to characterize quantitatively the different subsets of cells in a mesenchymal DFSC culture through High content imaging analysis. We speculate in the primary cellular cultures isolated from native tissues which have similar fibroblast-like morphology, different subsets of cells exist, and these different subsets are providing the multilineage potential of the said cultures.
Methods: Dental follicle of retained third molars were collected after routine extractions in Dental faculty (Medical university – Sofia) after informed consent was obtained from the patients. Pericoronal DFSC were seeded in 96 well plates and subdued to High-Content Cellular Analysis. The IN Cell Analyzer 6000 imaging system (GE Healthcare, Pittsburgh, USA) enabled identification and quantification of the cell markers expression. A total of 36 fields of view per well were photographed at x20 magnification. Cells were divided according to cell/background fluorescence intensity values measured by In Cell Analyzer work station 3.7.3 (x64) software (GE Healthcare): positive expression – fluorescence intensity >10; no expression/autofluorescence – fluorescence intensity <10.
Results:
We revealed that all selected markers are strongly expressed by approximately one third of the cell population. Therefore, nearly one third of the isolated cells are non-differentiated stem cells based on the amount of cells expressing stem cell markers. Of all the stem cell markers in the present study, CD90 was strongly expressed by the highest number of cells (49%). The expression of the following markers was found to be statistically significant when compared to control: CD90, CD117, vimentin, ALP, Sox9, CK10, p63, FGF7(KGF), Col3. The cells from each of the fractions may co-express markers from more than one group, as the cell culture is heterogeneous.
Conclusions: Primary DFSC cultures contain different subsets of cells, and these different subsets are providing the multilineage potential of the said cultures.