Isolation and co-culture of rat parenchymal and non-parenchymal liver cells to evaluate cellular interactions and response.
Isolation and co-culture of rat parenchymal and non-parenchymal liver cells to evaluate cellular interactions and response.
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DOI：
10.1038/srep25329
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发表时间：
2016-05-04
影响因子：
4.6
通讯作者：
Yarmush ML
中科院分区：
文献类型：
--
作者：
Bale SS;Geerts S;Jindal R;Yarmush ML
肝脏是人体的中枢器官，是宿主与外界环境之间的第一道防线。肝脏对任何外部干扰的反应是常驻肝细胞的集体反应。目前大多数体外肝脏模型都集中在肝细胞（主要代谢成分）上，忽略了周围环境和非实质细胞（NPC）的相互作用和提示。最近的研究表明，NPC的贡献是至关重要的，特别是在疾病条件下，药物及其代谢产物的结果。沿着肝细胞、NPCs-Kupffer（KC）、窦状内皮细胞（LSEC）和星状细胞（SC）是肝脏的主要细胞成分。在体外肝脏平台中掺入原代细胞对于模拟肝脏的功能及其总体反应至关重要。在此，我们从大鼠肝脏中分离单个NPC细胞组分，并将它们以transwell格式共培养，将原代大鼠肝细胞与LSEC，SC和KC合并。我们的研究结果表明，在共培养物中的多个细胞的存在和贡献捕获肝细胞和NPC之间的相互作用，并调节炎症刺激，如LPS的反应。分离和共培养方法可以为创建体外肝脏模型提供稳定的平台，所述体外肝脏模型提供除了单独的肝细胞之外的定义的功能。
The liver is a central organ in the human body, and first line of defense between host and external environment. Liver response to any external perturbation is a collective reaction of resident liver cells. Most of the current in vitro liver models focus on hepatocytes, the primary metabolic component, omitting interactions and cues from surrounding environment and non-parenchymal cells (NPCs). Recent studies suggest that contributions of NPCs are vital, particularly in disease conditions, and outcomes of drugs and their metabolites. Along with hepatocytes, NPCs–Kupffer (KC), sinusoidal endothelial (LSEC) and stellate cells (SC) are major cellular components of the liver. Incorporation of primary cells in in vitro liver platforms is essential to emulate the functions of the liver, and its overall response. Herein, we isolate individual NPC cell fractions from rat livers and co-culture them in a transwell format incorporating primary rat hepatocytes with LSECs, SCs, and KCs. Our results indicate that the presence and contributions of multiple cells within the co-culture capture the interactions between hepatocytes and NPC, and modulates the responses to inflammatory stimulus such as LPS. The isolation and co-culture methods could provide a stable platform for creating in vitro liver models that provide defined functionality beyond hepatocytes alone.