Detection and quantification of human-specific mRNA from hepatocyte-like cells derived from dental pulp using real-time polymerase chain reaction

Abstract

ObjectivesWe quantified viable hepatocyte-like cells (HLCs) administered via portal or tail veins in the livers and lungs of immunodeficient rats using real-time reverse transcription polymerase chain reaction (RT-PCR) and human glyceraldehyde 3-phosphate dehydrogenase (GAPDH) primers. MethodsImmunodeficient rats were infused with HLCs via portal or tail veins. mRNA was quantified based on the route of cell administration and the presence of liver injury. ResultsHuman-specific GAPDH mRNA primers detected 0.1 pg human RNA in 100 ng (1:106) of rat liver RNA. When infused into the portal vein, the quantity of HLC mRNA reduced to 5% 3 h after infusion. Most HLCs were entrapped in the lungs when infused via the tail vein and decreased to approximately 10% 6 h after infusion. A small number of HLCs made it to the liver but disappeared rapidly, regardless of liver injury. 24 h after infusion, viable HLCs were detected only in the lungs of rats with liver injury (P < 0.05). ConclusionsThe quantity of viable human cells in immunodeficient rats was estimated using real-time RT-PCR and primers specific to human mRNA.