A semiquantitative PCR technique for detecting chimerism in hamster-to-rat bone marrow xenotransplantation.
Although bone marrow transplantation has been used to induce donor-specific tolerance in many allogeneic models, similar effort in xenogeneic transplantation is met with obstacles like more severe graft versus host disease (GVHD). We are currently engaged in developing a GVHD-free hamster-to-rat xenotransplantation model using splenectomy, total body irradiation, and donor bone marrow transplantation. To test donor cell chimerism, particularly in the solid tissues, we developed a semiquantitative polymerase chain reaction (PCR) method using primers specific for hamster beta-actin and mitochondrial cytochrome C oxidase I and II (MCO I and II) genes and rat sex determination region on the Y chromosome (SRY) gene. Using this method, we estimated the level of hamster cells chimerism in rats subjected to splenectomy, total body irradiation (10 Gy), and hamster bone marrow transplantation (3 x 10(8) cell/recipient) and observed high levels of donor cells in all recipient tissues tested.
Kuddus
Ruhul H
RH
Thomas E Starzl Transplantation Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA. kuddusru@uvsc.edu
Lee
Y-H
YH
Valdivia
Luis A
LA
D000199:Actins / Q000235:genetics; D000818:Animals; D016026:Bone Marrow Transplantation / Q000276:immunology*; D006224:Cricetinae; D017931:DNA Primers; D003576:Electron Transport Complex IV / Q000235:genetics; D005260:Female; D024282:Genes, sry; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D012680:Sensitivity and Specificity; D013156:Splenectomy; D018183:Transplantation Chimera / Q000276:immunology*; D014183:Transplantation, Heterologous / Q000276:immunology*; D014916:Whole-Body Irradiation
D000199:Actins; D017931:DNA Primers; D003576:Electron Transport Complex IV