culturing pancreatic islets in microfluidic flow enhances morphology of the associated endothelial cells培养胰岛细胞在微流体流动增强相关的内皮细胞的形态.pdfVIP

Culturing Pancreatic Islets in Microfluidic Flow Enhances Morphology of the Associated Endothelial Cells 2. 1. 1 3 Krishana S. Sankar , Brenda J. Green , Alana R. Crocker , Jocelyne E. Verity , Svetlana M. Altamentova3, Jonathan V. Rocheleau1,2,3* 1 Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada, 2 Department of Physiology, University of Toronto, Toronto, Ontario, Canada, 3 Toronto General Research Institute, University Health Network, Toronto, Ontario, Canada Abstract Pancreatic islets are heavily vascularized in vivo with each insulin secreting beta-cell associated with at least one endothelial cell (EC). This structure is maintained immediately post-isolation; however, in culture the ECs slowly deteriorate, losing density and branched morphology. We postulate that this deterioration occurs in the absence of blood flow due to limited diffusion of media inside the tissue. To improve exchange of media inside the tissue, we created a microfluidic device to culture islets in a range of flow-rates. Culturing the islets from C57BL6 mice in this device with media flowing between 1 and 7 ml/24 hr resulted in twice the EC-density and -connected length compared to classically cultured islets. Media containing fluorescent dextran reached the center of islets in the device in a flow-rate-dependant manner consistent with improved penetration. We also observed deterioration of EC morphology using serum free media that was rescued by addition of bovine serum albumin, a known anti-apoptotic signal with limited diffusion in tissue. We further examined the effect of flow on beta-cells showing dampened glucose-stimulated Ca2+-response from cells at the periphery of the