Preservation of transporter function in 3D primary human hepatocyte cultures allows liver toxicity studies

Introduction: Primary human hepatocytes (hHeps) in 2D culture are the “gold standard” for in vitro toxicity studies. Since these conditions do not reflect tissue in vivo properties, hepatocytes rapidly lose their biotransformation capacity. To mimic in vivo conditions, we developed a 3D culture system maintaining hepatocytes polarity and function. Materials and methods: hHeps were isolated with two-step collagenase perfusion (according to ethical guidelines) and seeded in collagen I gels in 96-well-plates. As quality criteria for our system we measured cell viability, production of glucose and urea, functionalit…