Simultaneous Engraftment of Hepatocytes and Endothelial Cells in Mitomycin C Treated Fischer Rats

Abstract

Treatment with retrorsine crosslinks host hepatocyte DNA and prevents proliferation after partial hepatectomy (PH), allowing selective expansion of transplanted progenitors. Limitations of this protocol are the time required (7 weeks) and the carcinogenicity of retrorsine. This report describes a rapid transplantation protocol that replaces retrorsine with mitomycin C, a widely used chemotherapy agent. To promote donor cell expansion, dipeptidyl peptidase IV (DPPIV) deficient Fischer rats were injected with mitomycin C, and after 7 days, were injected following PH, with liver isolates containing both hepatocytes and endothelial cells from adult, fetal or newborn DPPIV+ Fischer rats. Injections of gentamicin initiated at one-day post surgery decreased mortality from 55% to 20%. By 4 weeks, host livers contained well-defined hepatocyte colonies that increased 2.6X in size between 4 and 9 weeks and expressed canalicular DPPIV and characteristic hepatocyte proteins. Host livers also contained large clusters of sinusoids lined with DPPIV+ donor cells positive for RECA-1, a pan-endothelial marker. Mixed colonies with donor hepatocytes and endothelial cells were also observed. Primary advantages of this protocol are the rapid production of donor colonies and the potential for developing a clinically applicable protocol for cellular therapy. Grants CA42715, CA93840 and P20RR017695.