Objective To investigate the cell compatibility of the porcine acellular lumens matrix substituting bile duct and evaluate the method to guide the clinical application of the porcine lumens scaffold. Methods Porcine bile duct and ureter were treated using detergent sodium dodecylsulphate (SDS) and 1% Triton X-100 to prepare the acellular lumens matrix. The toxic effects of different concentrations of acellular lumens matrix extract were tested by MTT to assess the proliferation of human scarfskin fibroblasts (HSF). The cytotoxicity of the target biomaterial was graded according to the national standards. The growth manner of the human intrahepatic bile duct endothelial cells (HIBDCs) seeded on the acellular lumens matrix was studied after 20 d under scanning electron microscopy.Results Acellular lumens matrix was completely devoid of cellular and nuclear material while maintaining the integrity of extracellular collagenous matrix. The cytotoxicity score of the matrix was in grade 0－1, which meant the biomaterial had no cytotoxicity. The microscopy showed the seeded HIBDCs had the potentials of spread and proliferation on the matrix, but there were few cells infiltrating into the acellular lumens matrix. Conclusions Porcine acellular lumens matrix is a natural non-toxic xenogenic lumens substitute with good cell affinity, but the time of adherence is long, so further endeavors are needed to improve the progress of adherence.