Vinken Mathieu, Elaut Greetje, Henkens Tom, Papeleu Peggy, Snykers Sarah, Vanhaecke Tamara, Rogiers Vera
Department of Toxicology, Vrije Universiteit Brussel, Belgium.
Methods Mol Biol. 2006;320:247-254. doi: 10.1385/1-59259-998-2:247.
Mimicking the in vivo microenvironment is one of the current strategies to maintain liver-specific functionality in primary cultured hepatocytes for long periods. Freshly isolated hepatocytes entrapped in collagen gel type I (collagen gel immobilization culture) or sandwiched between two layers of hydrated collagen type I (collagen gel sandwich culture) are known to display liver-specific functions (e.g., biotransformation capacity) for more than 6 wk. We describe how to set up both types of organotypical hepatocyte culture systems. Besides a detailed protocol, we give some practical tips, taken from our own experience with long-term hepatocyte culture.
模拟体内微环境是目前在原代培养肝细胞中长期维持肝脏特异性功能的策略之一。已知包埋于I型胶原凝胶(胶原凝胶固定培养)或夹于两层水合I型胶原之间(胶原凝胶夹心培养)的新鲜分离肝细胞可在超过6周的时间内表现出肝脏特异性功能(如生物转化能力)。我们描述了如何建立这两种类型的器官型肝细胞培养系统。除了详细的方案外,我们还根据自己长期肝细胞培养的经验给出了一些实用技巧。