@misc{LutterLiedtkeEckeetal., author = {Lutter, Anne-Helen and Liedtke, Victoria and Ecke, Annemarie and Scholka, Jenny and Muschter, Antje and Hentschel, Erik and Becker, Roland and Anderer, Ursula}, title = {Direct co-culture of human chondrogenic microtissues with osteoblast-like Saos-2 cells positively influence the differentiation potential of cartilage cells in 3D culture}, doi = {10.1007\%2Fs10354-018-0664-9.pdf}, pages = {S. 51}, language = {en} } @misc{LutterLiedtkeScholkaetal., author = {Lutter, Anne-Helen and Liedtke, Victoria and Scholka, Jenny and Muschter, Antje and Becker, Roland and Anderer, Ursula}, title = {Co-culture of human chondrogenic microtissues with osteoblast-like Saos-2 cells or HFF-1 fibroblasts influence the differentiation potential of spheroids}, series = {Journal of Cellular Biotechnology}, volume = {4}, journal = {Journal of Cellular Biotechnology}, number = {1/2}, issn = {2352-3697}, doi = {10.3233/JCB-189005}, pages = {31 -- 42}, abstract = {BACKGROUND: Tissue engineering has become a major field of research in biotechnology and biomedicine. As a consequence, cell-based therapeutic approaches are entering the hospitals, especially for skeletal regeneration. Traumatic injuries of cartilage are treated with autologous cell suspensions or in vitro generated cartilage tissues, but there is actually no therapy available for degenerative cartilage defects. However, Osteoarthritis (OA) is a major public health problem in the world affecting 240 million people globally. OBJECTIVE: To develop suitable in vitro tissues, the properties of chondrogenic spheroids should be optimized via coculture with cells naturally occurring as joint neighbours. METHODS: Human chondrocytes were isolated from condyles and propagated in monolayer culture. Scaffold-free spheroids were generated and co-cultured with joint-specific partner cells (osteoblast-like osteosarcoma cells, fibroblasts). Morphology and differentiation was analyzed using histochemistry (Alcian blue, Safranin O) and immunohistochemistry for cartilage markers (collagen type II, Sox9, proteoglycan), proliferation-associated protein (Ki67) and markers of connective tissue (collagen type I and actin). RESULTS: The provision of a more natural microenvironment in vitro via co-culture of chondrocyte-based aggregates with osteoblast-like Saos-2 cells enhanced the differentiation potential of chondrogenic spheroids towards hyaline cartilage. CONCLUSIONS: The study showed the positive influence of Saos-2 cells on the differentiation potential of human chondrocytes in co-culture.}, language = {en} } @misc{EckeScholkaHanschetal., author = {Ecke, Annemarie and Scholka, Jenny and Hansch, Anna and Becker, Roland and Anderer, Ursula}, title = {Human serum or FCS: Differentiation of human cartilage-like microtissues depends on serum selection}, series = {DGZ 2018 International Meeting of the German Society for Cell Biology: "Concepts of Cell Organisation and Dynamics"}, journal = {DGZ 2018 International Meeting of the German Society for Cell Biology: "Concepts of Cell Organisation and Dynamics"}, abstract = {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 f{\"u}r Wissenschaft, Forschung und Kultur", state of Brandenburg, Germany.}, language = {en} }