The experimental study of endothelial progenitor cells in rat liver decellularized scaffold

Objective To engineer intact whole rat liver decellularized scaffolds and repopulated bone marrow-derived endothelial progenitor cells (EPCs) into the scaffolds by continous perfusion technology thus providing experimental support for the vasculation of decellularized liver scaffolds in liver engineering. Methods Decellularized liver scaffolds were obtained by perfusing method. The composition and structure was examed by hematoxylin-eosin staining (HE) and immunohistochemistry. Bone marrow derived EPCs were characterised by immunofluorescence of CD31, CD133 and Vascular endothelial growth factor (VEGF). EPCs were recellularized into the decellularized scaffolds by portal-vein infusion method and cultured by the dynamic circulation perfusion device. After cultivation, HE staining and immunofluorescence of CD31 were conducted to observe the growth situation of EPCs in the scaffolds. Results The rat decellulariezd liver scaffolds were successfully obtained by perfusion method. Histological staining demonstrated the remove of cellular component and the reservation of extracellular cell matrix. Immunohistochemistry staining demonstrated the retention of collagen I and laminin. The primary cultured EPCs were postive for CD31, CD133 and VEGF. The circulation perfusion device was composed of a peristaltic pump, oxygenator, chamber and the convey tubes. HE staining and immunofluorence revealed that EPCs can be located around the blood vessel wall. Conclusion The primary cultured EPCs can grew around the blood vessel wall of the decellularized liver scaffolds by circulation perfusion method. Key words: Liver; Decellularized scaffolds; Circulation perfusion culture; Vascularization