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Isolation of PDPCs

XH Xin Huang
DH Donghua Huang
TZ Ting Zhu
XY Xiaohua Yu
KX Kaicheng Xu
HL Hengyuan Li
HQ Hao Qu
ZZ Zhiyuan Zhou
KC Kui Cheng
WW Wenjian Wen
ZY Zhaoming Ye
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PDPCs were harvested from the femurs of Sprague–Dawley (SD) rats (male, 4 weeks old). First, aseptically remove the muscle and connective tissue from the femurs. Next, the femurs were dissected, cut into small pieces (1–2 × 1–2 mm), and embedded in 5% low melting point agarose (Takara, Shiga, Japan) to prevent them from digestion. PDPCs were harvested by 0.2% collagenase digestion for 1 h at 37 °C. Single-cell suspensions were plated onto a 60-mm culture dish in Dulbecco’s Modified Eagle’s Medium–low glucose (DMEM, Gibco) with 10% (vol./vol.) fetal bovine serum (FBS; Gibco) and 1% (vol./vol.) penicillin/streptomycin (Gibco) at 37 °C with 5% CO2. The culture medium was changed every 3 days. The cells were used between passages 3 and 5.

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