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Optimization of condrocytes in 3D culture system for tissue engineering
Tissue Engineering, is also known as a field of Regenerative Medicine. Condrocytes 3D culture is necessary to obtain a sufficient number of cells for cartilage bioengineered in Bioprinter system. However, the current evidences demonstrate that cellular microenvironment direct cell behavior through influences in signaling pathways. Cells behave more like origin tissue when cultured in three-dimensional environments. The growth of cells in a 3D culture system can optimize uniform tissue spheroids which are chondrospheres or building blocks for fabrication cartilage which can be used in bioprinter in order to provide a natural environment by an additional dimension for external signals, receptor binding, ECM interactions and ability to design and develop the construction of organs and living microtissues with the deposition layer by layer through spheroids.
Objetivos - Metodologia - Resultados - Discussão dos Resultados/Objectives - Methodology - Results - Discussion of Results/Objetivos - Metodología - Resultados - Discusión de los resultados
Objective: Develop a 3D cell culture system for optimizing the cartilage bioengineered. Method: The auricular cartilage cells were cultured until the fourth passage. Agarose mold 2% through were made the mold Microtissue 3D Sigma Aldrich® and placed in 12 well plates , and later added the middle were added to 5x105 cells and maintained in culture until the fourteenth day, where they were centrifuged and posteriorly placed into microtubes 1,5ml marking the spheroids with Mitotracker to assess mitochondrial function, hoesth to assess the nucleus and alexxa fluor 488 to analyze the cytoskeleton formation when the cell dispose in sphere form. Results: The analyzes performed in fluorescence microscopy Zeiss® showed that after the formation of the spheroid core remained integrate the active mitochondrial activity and disposition of the cytoskeleton was observed very clearly due to condensation of the extracellular matrix that formed the spheroid. Conclusion: The cells placed in the mold agarose Microtissue 3D Sigma Aldrich® were agreggated up over the days and formed uniform and intact spheroids and demonstrate to be useful for further bioprinter application.
Considerações Finais/Final considerations/Consideraciones finales
Support - This work was supported by Univer.Fed of São Paulo, São Paulo, Brazil,CAPES,CNPq – Productivity Scholarship (SED 307389/2015-4 level 1D; MTD 306972/2015-8 level 2) FAPESP 2013/00288-4;2013/06877-1;INCT-Nat.Inst.Sci.andTech–Biofabris,CNPq,573661/2008-1,FAPESP08/57860-3;2010/20763-0,CNPQ 402958/2012-8.
Palavras-chave/Key words/Palabras clave
3D culture system; condrocytes; spheroids; tissue engineering
Celulas Tronco Pluripotentes
SILVIO EDUARDO DUAILIBI, Jennifer Adriane Santos, Vladimir Mironov, Rodrigo Alvarenga Rezende, Jorge Vicente Lopes Silva, Monica Talarico Duailibi