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[Realistic imaging of cell systems using confocal laser scanning microscopy exemplified by 3-dimensional chondrocyte
J Aigner1, E Wilmes, A Naumann
1Klinik und Poliklinik für Hals-Nasen- und Ohrenkranke, LMU München, Klinikum Grosshadem.
Background:
We have developed a three-dimensional model for tissue engineering of cartilage. Chondrocytes were isolated and first multiplied in conventional monolayer cultures. Then the cells are seeded with or without agarose on special absorbable scaffolds that provided stability and enabled three-dimensional cell distribution of the tissue-engineered cartilage. The aim of the study was to investigate the possibility of avoiding agarose in tissue engineering because of the potential risk of causing an inflammatory process in later human implantation.
Method:
For the first time we investigated cell distribution combined with vitality directly in the cell carrier under the conditions described by using confocal laser-scanning microscopy. Working with unfixed cells, this method enables the reconstruction of three-dimensional cell cultures with suitable cell markers that closely simulates the physiologic situation, thereby exceeding each other method.
Results:
It was evident that agarose had no positive effect on cell distribution and vitality.
Conclusion:
Further experiments concerning the effect of agarose on synthesis of cartilage-specific matrix are in progress.