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https://www.arca.fiocruz.br/handle/icict/24993
PROLIFERATION AND DIFFERENTIATION OF STEM CELLS IN CONTACT WITH ELUATE FROM FIBRIN-RICH PLASMA MEMBRANE
Author
Affilliation
Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.
Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.
Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.
Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.
Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.
Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.
Pontifícia Universidade Católica do Paraná. Curitiba, PR, Brasil.
Abstract
To evaluate the ability of the eluate from fibrin-rich plasma (FRP) membrane to induce proliferation and differentiation of isolated human adipose-derived stem cells (ASCs) into chondrocytes. How method: FRP membranes were obtained by centrifugation of peripheral blood from two healthy donors, cut, and maintained in culture plate wells for 48h to prepare the fibrin eluate. The SCATh were isolated from adipose tissue by collagenase digestion solution, and expanded in vitro. Cells were expanded and treated with DMEM-F12 culture, a commercial media for chondrogenic differentiation, and eluate from FRP membrane for three days, and labeled with BrdU for quantitative assessment of cell proliferation using the HighContent Operetta® imaging system. For the chondrogenic differentiation assay, the SCATh were grown in micromass for 21 days and stained with toluidine blue and aggrecan for qualitative evaluation by light microscopy. The statistical analysis was performed using ANOVA and Tukey’s test. The results were that there was a greater proliferation of cells treated with the eluate from FRP membrane compared to the other two treatments, where the ANOVA test showed significance (p < 0.001). The differentiation into chondrocytes was visualized by the presence of mucopolysaccharide in the matrix of the cells marked in blue toluidine and aggrecan. This way concluded in by treatment with eluate from FRP membrane stimulated cell proliferation and induced differentiation of the stem cells into chondrocytes, suggesting a potential application of FRP membranes in hyaline cartilage regeneration therapies.
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