Aim
To characterize stem cells originating from different dental tissues (apical papilla [SCAP], dental follicle [DFSC], and pulp [DPSC]) and test the capacity of Raman microspectroscopy to distinguish between the three dental stem cell types.
Methods
SCAP, DFSC, and DPSC cultures were generated from three immature wisdom teeth originating from three patients. Cell stemness was confirmed by inducing neuro-, osteo-, chondro-, and adipo-differentiaton and by mesenchymal marker expression analysis by flow-cytometry and real-time polymerase chain reaction. Cellular components were then evaluated by Raman microspectroscopy.
Results
We found differences between SCAP, DFSC, and DPSC Raman spectra. The ratio between proteins and nucleic acids (748/770), a parameter for discriminating more differentiated from less differentiated cells, showed significant differences between the three cell types. All cells also displayed a fingerprint region in the 600-700 cm
-1
range, and characteristic lipid peaks at positions 1440 cm
-1
and 1650 cm
-1
.
Conclusion
Although different dental stem cells exhibited similar Raman spectra, the method enabled us to make subtle distinction between them.
As data on the distribution of freshwater sponges (Porifera, Demospongiae, Spongillida) in Serbia are extremely scarce, we investigated the main Serbian rivers and lakes with respect to Porifera occurrence, for which 17 lotic and 11 lentic water bodies were selected. Sponges were found
in 11 of 17 rivers (62 specimens in total) and in 3 of 11 lakes/reservoirs (seven specimens in total). Classical morphological spicule analysis was coupled with polymerase chain reaction (PCR) and gene sequencing for species identification. Among the 69 collected speci- mens, five sponge species
of the family Spongillidae have been identified: Ephydatia fluviatilis (Linnaeus, 1759), Spongilla lacustris (Linnaeus, 1759), Ephydatia muelleri (Lieberkühn, 1856), Trochospongilla horrida Weltner, 1893, and Eunapius fragilis (Leidy, 1851).
The most frequently found sponge in Serbian rivers was E. fluviatilis (45% of all specimens), while the least frequent was E. fragilis (6 % of all specimens). The Tisa river has the highest sponge diversity (four species). In lentic water bodies, only E. fluviatilis (four
specimens) and S. lacustris (three specimens) were found. In general, sponges were infrequent and their abundance was low in Serbian fresh waters. While sponges seem to tolerate significant variations of physical and chemical parameters, some optimal values can be established.
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