TY - GEN A1 - Lutter, Anne-Helen A1 - Scholka, Jenny A1 - Richter, Heiko A1 - Anderer, Ursula T1 - Applying XTT, WST-1, and WST-8 to human chondrocytes: A comparison of membrane-impermeable tetrazolium salts in 2D and 3D cultures T2 - Clinical Hemorheology and Microcirculation KW - chondrogenic differentiation Y1 - 2017 U6 - https://doi.org/10.3233/CH-179213 SN - 1875-8622 SN - 1386-0291 VL - 67 IS - 3-4 SP - 327 EP - 342 ER - TY - GEN A1 - Lutter, Anne-Helen A1 - Hempel, Ute A1 - Anderer, Ursula A1 - Dieter, Peter T1 - Biphasic influence of PGE2 on the resorption activity of osteoclast-like cells derived from human peripheral blood monocytes and mouse RAW264.7 T2 - Prostaglandins, Leukotrienes and Essential Fatty Acids KW - osteoclastic differentiation Y1 - 2016 U6 - https://doi.org/10.1016/j.plefa.2016.03.017 SN - 1532-2823 SN - 0952-3278 VL - 111 SP - 1 EP - 7 ER - TY - GEN A1 - Lutter, Anne-Helen A1 - Hempel, Ute A1 - Dieter, Peter A1 - Anderer, Ursula T1 - Multinuclearity is not mandatory for Bone Resorption Mononuclear Osteoclasts are able to resorbe Extracellular Bone Matrix in vitro T2 - Bone-Tec, Munich, 2017 KW - mononuclear osteoclast Y1 - 2017 UR - https://www.researchgate.net/publication/327860900 ER - TY - GEN A1 - Lutter, Anne-Helen A1 - Scholka, Jenny A1 - Richter, Heiko A1 - Anderer, Ursula T1 - Comparison of membrane-impermeable tetrazolium salt assays in 2D and 3D cultures of human chondrocytes T1 - 5. Naturwissenschaftstag, Senftenberg, 2017 KW - WST-8, WST-1, XTT KW - 2D/3D cultures KW - chondrocyte Y1 - 2017 U6 - https://doi.org/10.13140/RG.2.2.10682.49600 ER - TY - GEN A1 - Lutter, Anne-Helen A1 - Hempel, Ute A1 - Dieter, Peter A1 - Anderer, Ursula T1 - The biomarker-based ODEM-assay affects cellular characteristics of osteoclasts in vitro Tissue culture polystyrene vs Osteoblast-Derived Extracellular Matrix T1 - 6th Munich Biomarker Conference, Munich, 2016 Y1 - 2016 U6 - https://doi.org/10.13140/RG.2.2.20195.96806 ER - TY - CHAP A1 - Lutter, Anne-Helen A1 - Hempel, Ute A1 - Dieter, Peter A1 - Anderer, Ursula T1 - Cost-efficient diagnostic platform to measure osteoclast activity T2 - 35. Jahrestagung der Deutschen Gesellschaft für Klinische Mikrozirkulation und Hämorheologie, Mainz, 2016 Y1 - 2016 U6 - https://doi.org/10.13140/RG.2.2.10550.19528 ER - TY - GEN A1 - Lutter, Anne-Helen A1 - Liedtke, Victoria A1 - Ecke, Annemarie A1 - Scholka, Jenny A1 - Muschter, Antje A1 - Hentschel, Erik A1 - Becker, Roland A1 - Anderer, Ursula T1 - Direct co-culture of human chondrogenic microtissues with osteoblast-like Saos-2 cells positively influence the differentiation potential of cartilage cells in 3D culture Y1 - 2018 U6 - https://doi.org/10.1007%2Fs10354-018-0664-9.pdf N1 - Austrian Bone Conference – ABC 2018, November 23–24, 2018, Vienna, Austria ER - TY - GEN A1 - Lutter, Anne-Helen A1 - Liedtke, Victoria A1 - Scholka, Jenny A1 - Muschter, Antje A1 - Becker, Roland A1 - Anderer, Ursula T1 - Co-culture of human chondrogenic microtissues with osteoblast-like Saos-2 cells or HFF-1 fibroblasts influence the differentiation potential of spheroids T2 - Journal of Cellular Biotechnology N2 - 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. Y1 - 2018 U6 - https://doi.org/10.3233/JCB-189005 SN - 2352-3697 SN - 2352-3689 VL - 4 IS - 1/2 SP - 31 EP - 42 ER - TY - GEN A1 - Ecke, Annemarie A1 - Lutter, Anne-Helen A1 - Scholka, Jenny A1 - Hansch, Anna A1 - Becker, Roland A1 - Anderer, Ursula T1 - Tissue Specific Differentiation of Human Chondrocytes Depends on Cell Microenvironment and Serum Selection T2 - Cells KW - Chondrocytes KW - Cartilage Y1 - 2019 U6 - https://doi.org/10.3390/cells8080934 SN - 2073-4409 VL - 8 IS - 8 ER - TY - GEN A1 - Lutter, Anne-Helen A1 - Muschter, Antje A1 - Anderer, Ursula T1 - Diagnostic Test System to detect resorption activity of osteoclasts in vitro just using small blood samples T2 - Osteologie KW - Osteoclast KW - diagnostic Y1 - 2019 U6 - https://doi.org/10.1055/s-0039-1680038 SN - 2567-5818 VL - 28 IS - 4 PB - Georg Thieme Verlag KG CY - Stuttgart ; New York ER - TY - GEN A1 - Lutter, Anne-Helen A1 - Ecke, Annemarie A1 - Scholka, Jenny A1 - Waldeck, Peter A1 - Tretter, Tim A1 - Anderer, Ursula T1 - Differentiation of human chondrocytes under the influence of microalgae-based exopolysaccharides T2 - 38. Jahrestagung der Deutschen Gesellschaft für Klinische Mikrozirkulation und Hämorheologie : Programm, 22. und 23. November 2019, Braunschweig N2 - BACKGROUND: Exopolysaccharides are formed by microalgae in response to adverse conditions and are found mainly as a coating around the cells. The protective function of EPS plays an important role in the survival of microalgae in polluted habitats. So far, the particular biological and physico-chemical properties of EPS have not been adequately investigated and their effect on cells such as human chondrocytes has not been tested. Microalgae in particular offer an ideal platform for the production of dietary supplements or drugs that enable new therapeutic approaches. METHODS: To investigate the influence of EPS on human chondrocytes, cytotoxicity and differentiation potential were analyzed in 3D cultures. RESULTS: We were able to demonstrate a positive, but donor-specific influence of EPS on the differentiation of human chondrocytes. Furthermore, the EPS showed no cytotoxic effects in a range of up to 250 µg EPS per ml medium. CONCLUSION: EPS have the potential to positively influence the differentiation of human chondrocytes. Further experiments are necessary to analyze the donor-specific effect. Y1 - 2019 UR - https://www.orbera.de/referenzen/DGKMH_2019/pdf_DGKMH_2019.pdf ER - TY - GEN A1 - Martin, Frank A1 - Neubert, Annemarie A1 - Lutter, Anne-Helen A1 - Scholka, Jenny A1 - Hentschel, Erik A1 - Richter, Heiko A1 - Anderer, Ursula T1 - MTS, WST-8, and ATP viability assays in 2D and 3D cultures: Comparison of methodologically different assays in primary human chondrocytes T2 - Clinical Hemorheology and Microcirculation N2 - BACKGROUND: Tissue engineering enables the production of three-dimensional microtissues which mimic naturally occurring conditions in special tissues. These 3D culture systems are particularly suitable for application in regenerative medicine or experimental pharmacology and toxicology. Therefore, it is important to analyse the cells in their 3D microenvironment with regard to viability and differentiation. Tetrazolium assays (WST-8 and MTS) are still the methods of choice for estimating the number of living, metabolically active cells, with WST-8 being cell-impermeable compared to MTS. In contrast to these methods, the ATP assay is an endpoint method based on the luciferase-induced reaction of ATP with luciferin after cell lysis. OBJECTIVE:We compared three methodologically different proliferation/toxicity assays (MTS, WST-8, ATP) in monolayer (2D) and 3D culture systems to improve the technically challenging determination of the number of viable cells. METHODS: Chondrocytes were isolated from human articular cartilage. Three different test systems (MTS, WST-8, ATP) were applied to monolayer cells (2D, varying cell numbers) and spheroids (3D, different sizes) in 96-well plates. The intracellular ATP concentration was determined by luciferase-induced reaction of ATP with luciferin using a luminometer. Formazan formation was measured spectrophotometrically after different incubation periods. Evaluation was performed by phase contrast microscopy (toxicity), correlation of cell count and ATP concentration or absorption signal (Gompertz function) and propidium iodide (PI) staining to proof the cell lysis of all cells in spheroids. RESULTS: In 2D culture, all three assays showed a good correlation between the number of seeded cells and the ATP concentration or absorption data, whereas the MTS-assay showed the lowest specificity. In 3D culture, the spheroid sizes were directly related to the number of cells seeded. The absorption data of the WST-8 and MTS assay correlated only for certain spheroid size ranges, whereas the MTS-assay showed again the lowest specificity. Only the measured intracellular ATP content showed a linear correlation with all spheroid sizes ranging from 100–1000 µm. The WST-8 assay revealed the second-best sensitivity which allows the measurement of spheroids larger than 240 µm. Phase contrast observation of monolayer cells showed toxic effects of MTS after 6 h incubation and no signs of toxicity of WST-8. Staining with propidium iodide showed complete lysis of all cells in a spheroid in the ATP assay. CONCLUSION: Among tetrazolium-based assays, WST-8 is preferable to MTS because of its non-toxicity and better sensitivity. When determining the number of viable cells in the 2D system, caution is advised when using the ATP assay because of its two-phase slope of the correlation graph concerning cell number and intracellular ATP. In 3D systems of human chondrocytes, the ATP-assay is superior to the other two test systems, as the correlation graph between cell number and intracellular ATP is biphasic. Since differentiation processes or other metabolic events can influence the results of proliferation and toxicity assays (determination of viable cells), this should be taken into account when using these test systems. KW - ATP assay KW - MTS KW - WST-8 KW - Human chondrocytes KW - Spheroid KW - 3D culture KW - 2D culture Y1 - 2024 UR - https://journals.sagepub.com/doi/abs/10.3233/CH-248101 U6 - https://doi.org/10.3233/CH-248101 SN - ISSN 1386-0291 VL - 88 SP - 3 EP - 19 PB - IOS Press ER -