Isolation and Culturing of Primary Murine Chondroprogenitor Cells : A Mammalian Model of Chondrogenesis

Vágó, Judit and Somogyi, Csilla and Takács, Roland and Barna, Krisztina Bíróné and Jin, Eun‐Jung and Zákány, Róza and Matta, Csaba (2024) Isolation and Culturing of Primary Murine Chondroprogenitor Cells : A Mammalian Model of Chondrogenesis. CURRENT PROTOCOLS, 4 (3). No. e1005. ISSN 2691-1299

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Abstract

Embryonic limb bud-derived micromass cultures are valuable tools for investigating cartilage development, tissue engineering, and therapeutic strategies for cartilage-related disorders. This collection of fine-tuned protocols used in our laboratories outlines step-by-step procedures for the isolation, expansion, and differentiation of primary mouse limb bud cells into chondrogenic micromass cultures. Key aspects covered in these protocols include synchronized fertilization of mice (Basic Protocol 1), tissue dissection, cell isolation, micromass formation, and culture optimization parameters, such as cell density and medium composition (Basic Protocol 2). We describe techniques for characterizing the chondrogenic differentiation process by histological analysis (Basic Protocol 3). The protocols also address common challenges encountered during the process and provide troubleshooting strategies. This fine-tuned comprehensive protocol serves as a valuable resource for scientists working in the fields of developmental biology, cartilage tissue engineering, and regenerative medicine, offering an updated methodology for the study of efficient chondrogenic differentiation and cartilage tissue regeneration.

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