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Pharmacogenetic selection of transplanted human hepatocytes in immunocompetent rats
Author(s) -
Smolic Martina,
Wu Catherine H,
Madadi Shilpa,
Gupta Nidhi,
Smolic Robert,
Coash Marcy,
Smith Jeanette,
Wu George Y
Publication year - 2012
Publication title -
journal of digestive diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.684
H-Index - 51
eISSN - 1751-2980
pISSN - 1751-2972
DOI - 10.1111/j.1751-2980.2012.00632.x
Subject(s) - medicine , pharmacogenetics , selection (genetic algorithm) , pharmacology , computational biology , genetics , biology , genotype , artificial intelligence , gene , computer science
Objective To introduce a genetic survival advantage for transplanted human hepatocytes over host cells in rats. Methods Green fluorescent protein ( GFP ) was introduced into H uh‐7 human hepatoma cells to create fluorescent GFP ‐ H uh‐7 cells. mRNA of CYP 2 E 1, the enzyme that converts acetaminophen ( APA ) into hepatotoxic intermediates, was quantified by real‐time polymerase chain reaction ( PCR) . The effects of APA on GFP ‐ H uh‐7 and control H uh‐7 cells were determined in a cell culture. Immunological tolerance was induced by the injection of GFP ‐ Huh ‐7 cells into fetal rats in utero . The GFP ‐ H uh‐7 cells were transplanted after birth of the rats into tolerant rats followed by APA treatment. Serum alanine aminotransferase ( ALT ) levels and liver histological data were obtained. GFP ‐ H uh‐7 cells were detected by quantitive PCR and microscopy. Results CYP 2 E 1 mRNA levels in the GFP ‐ H uh‐7 cells were 2.7% of parental H uh‐7 cells. In 1 mmol/ L APA , parental H uh‐7 cells decreased by 60% while GFP ‐ H uh‐7 cells increased to within 95% of untreated controls after 5 days. In rats in which GFP ‐ H uh‐7 cells were transplanted and treated with APA , serum ALT increased to a peak of 200 U / L on day 1 and returned to normal levels by day 3. Fluorescence microscopy of liver specimens from rats transplanted with GFP ‐ H uh‐7 cells showed substantial increases in GFP ‐ H uh‐7, but not H uh‐7 cells by day 7 after APA treatment. Real‐time PCR confirmed a 10‐fold increase of GFP mRNA in APA ‐treated rats, but not in those without APA treatment. Conclusions The difference in CYP 2 E 1 gene expression between GFP ‐ H uh‐7 and rat hepatocytes provides a convenient means for the enrichment of transplanted human cells in rat liver.