Research Day
CONTROLLED-ASSEMBLY OF CHONDROCYTE SPHEROIDS USING 3D PRINTED MOLDS
Document Type
Abstract
Date
2021
Abstract
Spheroids are becoming popular in tissue engineering due to their ability to better mimic the 3D architecture of living tissues compared to 2D-cell cultures. Spheroids have granted new insights in personalized medicine, disease pathogenesis, drug discovery, and regenerative medicine as potential alternatives for cell and even whole-organ transplantation. However, the fabrication processes of spheroids have not been perfected and there are many procedural techniques that need to be optimized. The purpose of this study is to develop a procedure to consistently create chondrocyte spheroids of a desired size using agarose molds. The agarose is designed to contain 800-m diameter wells using 3D printed negative molds (NM). 3D models of NM were produced using FreeCAD software and printed using DLP 3D printer. Excess resin was cleaned and NM were cured under UV-light. Next, sterile agarose solution (2%) was prepared and pipetted into the NM. Hardened molds were removed and washed with culture medium. Different concentrations of human chondrocytes (up to 5.105 cell/well) were plated onto the agarose molds and incubated overnight to form spheroids. Finally, spheroids were transferred on non-adhering culture plates and cultured for up to 4-weeks. Spheroids were harvested, stained using DAPI and visualized under a fluorescence microscope. Spheroids shape (circularity) and size were determined by image analysis.