TY - GEN A1 - Ecke, Annemarie A1 - Scholka, Jenny A1 - Hansch, Anna A1 - Becker, Roland A1 - Anderer, Ursula T1 - Human serum or FCS: Differentiation of human cartilage-like microtissues depends on serum selection T2 - DGZ 2018 International Meeting of the German Society for Cell Biology: “Concepts of Cell Organisation and Dynamics” N2 - Introduction: Regeneration of articular cartilage defects is limited. Hence, cell-based therapies for the treatment were developed using e.g. chondrocytes isolated from patient biopsies. For cell culture fetal calf serum (FCS) is often used as media supplement to establish engineering processes for transplants in regenerative medicine. However, the usage of xenogeneic serum could mask the intrinsic behavior of human cells in vitro. This could be particularly important for the dedifferentiation process during cell expansion and redifferentiation in 3D-culture. The aim of this study was to compare the development of microtissues derived from chondrocytes cultivated in presence of FCS with those cultivated in human serum (HS). Material and Methods: Human articular chondrocytes isolated from knee joints were expanded in medium with HS or FCS. Scaffold-free microtissues were generated using the agar overlay technique. After four and eight weeks the macroscopic appearance (reflected-light microscopy) was documented and the size of the microtissues was determined. The differentiation degree was evaluated via histology to visualize typical glycosamino¬glycans (Safranin O, Alcian blue) and immunohisto¬chemistry (IH) to detect cartilage-specific markers (collagen type II, proteoglycans, COMP, Sox9) on cryosections. Results and Conclusion: Cells from all donors in both medium compositions generated microtissues – but with distinct differences in the differentiation levels. Microtissues cultured in medium with FCS showed a higher differentiation level compared to microtissues in HS. This is evidenced by a higher amount of matrix resulting in bigger microtissues (diameter in FCS is 42% larger compared to HS) and a more cartilage-like composition of the matrix (not any/little colouring in HS versus intense staining in FCS). IH analysis showed the same pattern. Not any/less positivity for collagen type II, proteoglycan, COMP, and Sox9 in HS versus moderate-to-high positivity in FCS-cultured microtissues. Absolute values vary from donor to donor whereas an individualized expression profile only became visible in microtissues cultured in HS medium. HS should be preferred for the engineering of cartilage-like microtissues, as it rather enables a "human-based" situation in vitro. Supported by „Gesundheitscampus Brandenburg“ and „Ministerium für Wissenschaft, Forschung und Kultur“, state of Brandenburg, Germany. KW - cartilage-like microtissues KW - serum type Y1 - 2020 UR - https://opus4.kobv.de/opus4-UBICO/frontdoor/index/index/docId/25239 UR - https://www.researchgate.net/publication/338863739_HUMAN_SERUM_OR_FCS_DIFFERENTIATION_OF_HUMAN_CARTILAGE-LIKE_MICROTISSUES_DEPENDS_ON_SERUM_SELECTION ER -