The goal of this project is to compare chondrogenic differentiation potential of various cultivated humans MSCs (from Bone Marrow, Adipose Tissue and Umbilical Cord). The cells are cultivated in chondrogenic culture media supplemented by TGF-β3 and human PRP. The cell viability is measured using MTS assay. The MTS substrate is reduced by cellular succinate dehydrogenase and the absorbance change is determined spectrophotometrically. The cell proliferation is assessed by measuring the content of dsDNA synthesized on the scaffold. The scaffolds are treated with cell lysis buffer and the amount of the released dsDNA is determined using a fluorescence probe. Confocal microscopy is used to visualize the cells. The cell morphology of fixated cells is visualized using DiOC-6 and propidium iodide staining. The viable cells are visualized by BCECF-AM. The dead cells are visualized by propidium iodide.
Chondrogenic potential of the cells will be assessed via immunohistochemistry to visualize cartilage matrix proteins synthesized on the scaffold and by determination of gene expression of selected specific chondrogenic markers (such as COL II, aggrecan and COX9) by PCR.
Click here to see summary of first results - Chondroveil_Dr._Tiegermann.pdf