Detection and quantification of human-specific mRNA from hepatocyte-like cells derived from dental pulp using
Naohiro Shimamura1, Kazuyuki Fujii2, Shogo Ohkoshi3
1Clinical Examination, The Nippon Dental University Graduate School of Life Dentistry at Niigata, Niigata, Japan.
Objectives:
We 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.
Methods:
Immunodeficient 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.
Results:
Human-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).
Conclusions:
The quantity of viable human cells in immunodeficient rats was estimated using real-time RT-PCR and primers specific to human mRNA.
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