Transfection of Primary Hepatocytes with Liver-Enriched Transcription Factors Using Adenoviral Vectors.
Primary cultured hepatocytes are probably the best model to study endogenous metabolic pathways, toxicity, or drug metabolism. Many of these studies require expression of ectopic genes. It would be desirable to use a method of transfection that allows dose-response studies, high efficiency of transfection, and the possibility to express several genes at the same time. Adenoviral vectors fulfill these requirements, becoming a valuable tool for primary hepatocyte transfection. Moreover, they are easy to generate and do not require a high level of biocontainment. In the present chapter, we describe the generation, cloning, amplification, and purification of an adenoviral vector capable of infecting primary cultured hepatocytes. This recombinant adenovirus induces robust expression of the protein of interest in hepatocytes within a wide range of doses.
Unidad de Hepatología Experimental-CIBERehd, Instituto de Investigación Sanitaria La Fe, Valencia, Spain.
Marta
M
Benet
Ramiro
R
Jover
Roque
R
Bort
D000256:Adenoviridae / Q000235:genetics*; D003001:Cloning, Molecular; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D061251:Primary Cell Culture / Q000379:methods; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic; D014162:Transfection / Q000379:methods*
D014157:Transcription Factors