DIFFERENTIATION OF MESENCHYMAL LIMB BUD CELLS TO CHONDROCYTES IN ALGINATE BEADS |
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| Authors: | M SHAKIBAEI P De SOUZA |
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| Institution: | 1. University Grenoble Alpes, Faculty of Medicine, Laboratory AGEIS EA 7407, Team Tools for e-Gnosis Medical, F-38700 La Tronche, France;2. The National Natural History Collections, The Hebrew University of Jerusalem, 9190401 Jerusalem, Israel;1. Rheumatology Department, Hospital Universitario de La Princesa, IIS-IP, Madrid, Spain;2. Rheumatology Department, Hospital Universitario de San Pedro, Logroño, Rioja, Spain;3. Radiology Department, Hospital Universitario de La Princesa, Madrid, Spain;1. Molecular Medicine Centre, Medical University of Sofia, 1431 Sofia, Bulgaria;2. Department of Medical Chemistry and Biochemistry, Medical Faculty, Medical University of Sofia, 1431 Sofia, Bulgaria;3. Clinic of Pediatrics, University Hospital “Alexandrovska”, 1431 Sofia, Bulgaria;4. Imaging Department, University Hospital “St Ivan Rilski”, 1431 Sofia, Bulgaria;5. National Genetic Laboratory, University Hospital of Obstetrics and Gynecology, 1431 Sofia, Bulgaria;1. Cardiac Intensive Care Unit, Heart Center, Guangzhou Women and Children''s Medical Center, Guangzhou Medical University, Guangzhou, China;2. The First Affiliated Hospital of Jinan University, Guangzhou, Jinan University, Guangzhou, China;3. Institute of Pediatrics, Guangzhou Women and Children''s Medical Center, Guangzhou Medical University, Guangzhou, China;4. Department of Pediatric Surgery, Guangzhou Women and Children''s Medical Center, Guangzhou Medical University, Guangzhou, China;1. Department of cardiology, Thorax Center, Erasmus MC, Rotterdam, The Netherlands;2. Department of Clinical Chemistry, Erasmus MC, Rotterdam, The Netherlands |
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| Abstract: | Manyin vitromodels of embryonic material used for the cultivation of chondrocytes yield mixed cultures consisting of chondrocytes and fibroblast-like cells. For the optimization of cartilage cell cultures, alginate, a semisolid medium, was employed to obtain pure chondrocyte cultures. Isolated mesenchymal cells from 12-day-old mouse limb buds were grown in alginate for up to 4 weeks. A sub-population of the cells differentiated to chondrocytes and exhibited a stable phenotype until the end of the culture period. After 3 to 4 days a cartilage-specific matrix started to develop. Fibroblast-like cells from this mixed culture did not survive; they became necrotic. When alginate was later on dissolved by chelating agents, only chondrocytes were isolated. During dissolution of alginate and centrifugation, chondrocytes did not lose their contact with their new matrix present on their surfaces. Cultivation of these chondrocytes or chondrones in mass culture yields a pure chondrocyte population. Immunoelectron microscopic investigations revealed collagen type II, fibronectin, decorin and chondroitin sulfate-proteoglycans in the chondrocyte capsules and in mass culture. |
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| Keywords: | limb bud cells alginate culture chondrocyte differentiation organoid culture |
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