CONFOCAL LASER-SCANNING MICROSCOPY ENABLE S REALISTIC RECONSTRUCTION OF CELL SYSTEMS AS ILLUSTRATED WITH 3-DIMENSIONAL CULTURED CHONDROCYTES

Background: We have developed a three-dimensional model for tissue eng ineering 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 stabili ty and enabled three-dimensional cell distribution of the tissue-engin eered cartilage. The aim of the study was to investigate the possibili ty of avoiding agarose in tissue engineering because of the potential risk of causing an inflammatory process in later human implantation. M ethod: For the first time we investigated cell distribution combined w ith vitality directly in the cell carrier under the conditions describ ed by using confocal laser-scanning microscopy. Working with unfixed c ells, this method enables the reconstruction of three-dimensional cell cultures with suitable cell markers that closely simulates the physio logic situation, thereby exceeding each other method. Results: It was evident that agarose had no positive effect on cell distribution and v itality. Conclusion: Further experiments concerning the effect of agar ose on synthesis of cartilage-specific matrix are in progress.