Then, saline, 5×104, 2×105, or 1×106 freshly isolated (no expansion) and expanded CD34+ cells/kg body weight were transplanted via spleen, respectively. The administration of CCl4 was continued for three more weeks until the rats were sacrificed. Examination items were as follows. 1)FACS, real-time PCR and gene array analysis of freshly isolated and expanded CD34+ cells, 2) morphometry Ku-0059436 price of fibrotic areas in Azan-Mallory stained liver, 3) immunohistochemistry using anti-α-smooth

muscle actin (SMA), CD31, keratin19, albumin and PCNA antibodies, and 4) the expression of metalloproteinase and tissue inhibitor of metalloproteinase-1 by gelatin zymography and real-time PCR. Results: For 7 days in culture, CD34+ cells were effectively expanded to 8-fold. Increased expression of VE-cadherin, KDR and Tie2 was determined by FACS analysis. The expression of VEGF, transforming growth factor-α, fibroblast growth factor-2, endothelial nitric oxide synthase and angiopoietin-2

in expanded CD34+ cells was increased compared with that in freshly isolated CD34+ cells. Gene array analysis showed that the most up-regulated gene in expanded CD34+ cells compared with freshly isolated CD34+ cells was integrin-3β. Expanded CD34+ cell transplantation reduced liver fibrosis with the decrease of αSMA positive cells. The transplanted cells differentiated into CD31+ and smooth myosin heavy chain-1+ cells. The transplantation of expanded CD34+ cells significantly up-regulated the number of PCNA positive hepatocyte compared with the transplantation of freshly isolated CD34+ in 3 different groups of cell number, respectively. Conclusion: Torin 1 ic50 These observations suggest that ex vivo expanded CD34+ cell transplantation may become a promising therapeutic strategy for patients with decompensated liver cirrhosis. Disclosures: Michio Sata – Speaking and Teaching: MSD K. K., Chugai 6-phosphogluconolactonase Pharmaceutical Co., The following people have nothing to disclose:

Toru Nakamura, Takuji Torimura, Hiroshi Masuda, Hideki Iwamoto, Hironori Koga, Mitsuhiko Abe, Yu Ikezono, Osamu Hashimoto, Takato Ueno Hepatocyte transplantation is a potential treatment for a myriad of liver disorders that are currently only curable by liver transplantation. A major limiting factor of hepatocyte transplantation is an inability to non-invasively and longitudinally monitor engraftment and expansion of transplanted cells. We hypothesized that the sodium iodide symporter (NIS) gene could be used to visualize transplanted hepatocytes and set out to test this reporter system in a rodent model of liver repopulation. FAH+ C57BI/6J mouse hepatocytes were transduced ex vivo using a lentiviral vector containing the mouse Slc5a5 (NIS) gene under the control of a liver specific promoter. Transduction efficiencies of 70-80% were achieved and NIS-labeled cells could robustly concentrate radiolabeled iodine in vitro.