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Primary chondrocyte isolation and culture

CW Cuicui Wang
JY Jun Ying
XN Xiaolei Nie
TZ Tianhong Zhou
DX Ding Xiao
GS Gaurav Swarnkar
YA Yousef Abu-Amer
JG Jianjun Guan
JS Jie Shen
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Primary chondrocytes were isolated from the ribcages of neonatal C57BL/6J pups as previously described.56 Briefly, the ribcages were dissected without soft tissue, followed by 2 mg·mL−1 pronase (Millipore Sigma, #10165921001) and 3 mg·mL−1 collagenase D (Millipore Sigma, #11088866001) digestion with agitation. The remaining sterna were further digested with 3 mg·mL−1 collagenase D for 4 to 6 h. Chondrocytes were collected and cultured in DMEM with 10% FBS. Following 48 h of treatment with vehicle and 1 ng·mL−1 IL-1β, the culture medium was collected for the angiogenic protein array and HUVEC angiogenesis assay.

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