Aleksandra Policha1, Ping Zhang1, Bing Shue2, Amanda Smolock2, Lily Chang2, Masaya Jimbo2, Thomas Tulenko3, Neil Moudgill4, Anumeha Whisenhunt4, Atul Rao4, Joshua Eisenberg4, Paul DiMuzio4
1Thomas Jefferson University Hospital, Department of General Surgery, Philadelphia, PA; 2Jefferson Medical College, Philadelphia, PA; 3Cooper University Hospital, Camden, NJ; 4Thomas Jefferson University Hospital, Department of General Surgery, Division of Vascular and Endovascular Surgery, Philadelphia, PA.
OBJECTIVES: Surgical bypass and dialysis access creation are common procedures performed in diabetic patients. Only 45% of patients have autologous vascular tissue available for use as a conduit for these procedures. We have developed an alternative conduit composed of a vascular scaffold seeded with autologous adipose-derived stem cells (ASC) differentiated into endothelial cells (EC) to create a non-thrombogenic lumen. Herein, we examine ASC from diabetic (DM) vs. non-diabetics (NDM) patients in terms of isolation efficiency, proliferation, commitment towards EC lineage, and seeding onto a vascular scaffold.
METHODS: ASC were isolated from peri-umbilical liposuction specimens by collagenase dispersion (DM n=53; NDM n=145). Isolation efficiency was defined by number of ASC isolated per gram of adipose tissue. Proliferation was assessed by constructing growth curves over 14d. ASC were differentiated in endothelial growth medium for 3wk. Differentiation was determined by measuring EC-specific gene expression (CD31, vWF) using qPCR. Lastly, ASC were seeded onto decellularized vein, flow conditioned from 0-9 dynes over 5d, and examined using confocal microscopy.
RESULTS: ASC isolation efficiency did not differ between DM and NDM patients (224,028 cells/gm vs. 259,345 cells/gm, respectively; p=0.21). The growth curves for DM (n=6) and NDM (n=6) ASC also appeared similar, with no significant differences observed in cells counts between days 1-14. After 1, 2, and 3wk in culture, no significant difference in CD31 or vWF expression between DM (n=6) and NDM (n=6) cells was observed. Finally, retention of ASC to vascular scaffolds under physiological shear stress was similar between the two groups.
CONCLUSIONS: ASC are isolated from diabetics in quantities similar to non-diabetics. These cells grow at similar rates, exhibit differentiation into an EC phenotype, and line the lumen of a tissue engineered vascular graft.
AUTHOR DISCLOSURES: L. Chang, Nothing to disclose; P. DiMuzio, Nothing to disclose; J. Eisenberg, Nothing to disclose; M. Jimbo, Nothing to disclose; N. Moudgill, Nothing to disclose; A. Policha, Nothing to disclose; A. Rao, Nothing to disclose; B. Shue, Nothing to disclose; A. Smolock, Nothing to disclose; T. Tulenko, Nothing to disclose; A. Whisenhunt, Nothing to disclose; P. Zhang, Nothing to disclose.
Posted April 2012