Quantification of chondrogenic differentiation in monolayer cultures of mesenchymal stromal cells. 2019

Tim Ruhl, and Justus P Beier
Department of Plastic Surgery, Hand Surgery - Burn Center, University Hospital RWTH Aachen, Germany. Electronic address: truhl@ukaachen.de.

Determining inhibitory or supportive effects of biological, chemical or physical factors on cell fates, including chondrogenic differentiation of mesenchymal stromal cells (MSCs), requires quantification techniques that are rapid, reproducible, and able to monitor these effects over time. Methods currently used to analyze chondrogenic differentiation are either qualitative staining procedures or indirect DNA quantifications. Because of these limitations, further methods are needed to improve determination of chondrogenic differentiation. In the present study, we applied a histological staining method, which is established for investigation of articular cartilage degeneration by use of Safranin O dye, on chondrogenic differentiated cells in monolayer cultures. MSCs were differentiated on 12-well formats, at increasing concentrations of TGF-β3, cell numbers, and incubation times. Quantification was performed by solubilizing the adsorbed dye into isopropanol followed by determining the optical density (O.D.) through spectrophotometry. Our results show that the O.D. is directly related to cell numbers and incubation periods, and that the technique is applicable to study agents which affect chondrogenic differentiation.

UI MeSH Term Description Entries
D010619 Phenazines
D002452 Cell Count The number of CELLS of a specific kind, usually measured per unit volume or area of sample. Cell Density,Cell Number,Cell Counts,Cell Densities,Cell Numbers,Count, Cell,Counts, Cell,Densities, Cell,Density, Cell,Number, Cell,Numbers, Cell
D002454 Cell Differentiation Progressive restriction of the developmental potential and increasing specialization of function that leads to the formation of specialized cells, tissues, and organs. Differentiation, Cell,Cell Differentiations,Differentiations, Cell
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D013053 Spectrophotometry The art or process of comparing photometrically the relative intensities of the light in different parts of the spectrum.
D059630 Mesenchymal Stem Cells Mesenchymal stem cells, also referred to as multipotent stromal cells or mesenchymal stromal cells are multipotent, non-hematopoietic adult stem cells that are present in multiple tissues, including BONE MARROW; ADIPOSE TISSUE; and WHARTON JELLY. Mesenchymal stem cells can differentiate into mesodermal lineages, such as adipocytic, osteocytic and chondrocytic. Adipose Tissue-Derived Mesenchymal Stem Cell,Adipose Tissue-Derived Mesenchymal Stromal Cell,Adipose-Derived Mesenchymal Stem Cell,Bone Marrow Mesenchymal Stem Cell,Mesenchymal Stromal Cell,Mesenchymal Stromal Cells,Multipotent Bone Marrow Stromal Cell,Multipotent Mesenchymal Stromal Cell,Adipose Tissue-Derived Mesenchymal Stem Cells,Adipose Tissue-Derived Mesenchymal Stromal Cells,Adipose-Derived Mesenchymal Stem Cells,Adipose-Derived Mesenchymal Stromal Cells,Bone Marrow Mesenchymal Stem Cells,Bone Marrow Stromal Cell,Bone Marrow Stromal Cells,Bone Marrow Stromal Cells, Multipotent,Bone Marrow Stromal Stem Cells,Mesenchymal Progenitor Cell,Mesenchymal Progenitor Cells,Mesenchymal Stem Cell,Mesenchymal Stem Cells, Adipose-Derived,Mesenchymal Stromal Cells, Multipotent,Multipotent Bone Marrow Stromal Cells,Multipotent Mesenchymal Stromal Cells,Stem Cells, Mesenchymal,Wharton Jelly Cells,Wharton's Jelly Cells,Adipose Derived Mesenchymal Stem Cell,Adipose Derived Mesenchymal Stem Cells,Adipose Derived Mesenchymal Stromal Cells,Adipose Tissue Derived Mesenchymal Stem Cell,Adipose Tissue Derived Mesenchymal Stem Cells,Adipose Tissue Derived Mesenchymal Stromal Cell,Adipose Tissue Derived Mesenchymal Stromal Cells,Mesenchymal Stem Cells, Adipose Derived,Progenitor Cell, Mesenchymal,Progenitor Cells, Mesenchymal,Stem Cell, Mesenchymal,Stromal Cell, Mesenchymal,Stromal Cells, Mesenchymal,Wharton's Jelly Cell,Whartons Jelly Cells
D018929 Cell Culture Techniques Methods for maintaining or growing CELLS in vitro. Cell Culture,Cell Culture Technique,Cell Cultures,Culture Technique, Cell,Culture Techniques, Cell
D020219 Chondrogenesis The formation of cartilage. This process is directed by CHONDROCYTES which continually divide and lay down matrix during development. It is sometimes a precursor to OSTEOGENESIS.
D064368 Healthy Volunteers Persons with no known significant health problems who are recruited to participate in research to test a new drug, device, or intervention as controls for a patient group. (from http://clinicalcenter.nih.gov/recruit/volunteers.html, accessed 2/14/2013) Healthy Participants,Healthy Subjects,Human Volunteers,Normal Volunteers,Healthy Participant,Healthy Subject,Healthy Volunteer,Human Volunteer,Normal Volunteer,Participant, Healthy,Participants, Healthy,Subject, Healthy,Subjects, Healthy,Volunteer, Healthy,Volunteer, Human,Volunteer, Normal,Volunteers, Human,Volunteers, Normal

Related Publications

Tim Ruhl, and Justus P Beier
January 2021, Methods in molecular biology (Clifton, N.J.),
Tim Ruhl, and Justus P Beier
January 2023, Methods in molecular biology (Clifton, N.J.),
Copied contents to your clipboard!