RESUMEN
In vitro studies of drug toxicity and drug-drug interactions are crucial for drug development efforts. Currently, the utilization of primary human hepatocytes (PHHs) is the de facto standard for this purpose, due to their functional xenobiotic response and drug metabolizing CYP450 enzyme metabolism. However, PHHs are scarce, expensive, require laborious maintenance, and exhibit lot-to-lot heterogeneity. Alternative human in vitro platforms include hepatic cell lines, which are easy to access and maintain, and induced pluripotent stem cell (iPSC) derived hepatocytes. In this study, we provide a direct comparison of drug induced CYP3A4 and PXR expression levels of PHHs, hepatic cell lines Huh7 and HepG2, and iPSC derived hepatocyte like cells. Confluently cultured Huh7s exhibited an improved CYP3A4 expression and were inducible by up to 4.9-fold, and hepatocytes differentiated from human iPSCs displayed a 3.3-fold CYP3A4 induction. In addition, an increase in PXR expression levels was observed in both hepatic cell lines and iPSC derived hepatocytes upon rifampicin treatment, whereas a reproducible increase in PXR expression was not achieved in PHHs. Our results indicate that both hepatoma originated cell lines and iPSCs may provide alternative sources to primary hepatocytes, providing reliable and reproducible results for CYP3A4/PXR metabolism, upon in vitro maturation. This study may serve as a guide for the selection of suitable and feasible in vitro platforms for drug-drug interaction and toxicology studies.
Asunto(s)
Inductores del Citocromo P-450 CYP3A/farmacología , Citocromo P-450 CYP3A/metabolismo , Hepatocitos/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacos , Diferenciación Celular , Línea Celular , Evaluación Preclínica de Medicamentos/métodos , Interacciones Farmacológicas , Hepatocitos/fisiología , Humanos , Células Madre Pluripotentes Inducidas/fisiología , Receptor X de Pregnano/metabolismo , Reproducibilidad de los Resultados , Pruebas de Toxicidad/métodosRESUMEN
IMPACT STATEMENT: In this review, we address the potential of human-induced pluripotent stem cell-based hepatobiliary differentiation technology as a means to study human liver development and cell fate determination, and to model liver diseases in an effort to develop a new human-relevant preclinical platform for drug development.
Asunto(s)
Conductos Biliares/efectos de los fármacos , Desarrollo de Medicamentos/métodos , Células Madre Pluripotentes Inducidas/efectos de los fármacos , Hepatopatías/terapia , Hígado/efectos de los fármacos , Proteínas Adaptadoras Transductoras de Señales/fisiología , Adulto , Animales , Conductos Biliares/citología , Diferenciación Celular , Línea Celular , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Evaluación Preclínica de Medicamentos/métodos , Cuerpos Embrioides/efectos de los fármacos , Proteínas Hedgehog/fisiología , Hepatocitos/efectos de los fármacos , Humanos , Técnicas In Vitro , Células Madre Pluripotentes Inducidas/citología , Hígado/citología , Hepatopatías/patología , Modelos Animales , Transducción de Señal/efectos de los fármacos , Factores de Transcripción/fisiología , Proteínas Señalizadoras YAPRESUMEN
The recent advances in the induced pluripotent stem cell (iPSC) research have significantly changed our perspectives on regenerative medicine by providing researchers with a unique tool to derive disease-specific stem cells for study. In this review, we describe the human iPSC generation from developmentally diverse origins (i.e. endoderm-, mesoderm-, and ectoderm- tissue derived human iPSCs) and multistage hepatic differentiation protocols, and discuss both basic and clinical applications of these cells including disease modeling, drug toxicity screening/drug discovery, gene therapy and cell replacement therapy.