Long Term Maintenance of Functional Primary Hu-man Hepatocytes in Organoid Culture Using Natural Mi-croparticles

Background: To discover new drugs in clinical trials, extensive preclinical testing has to be done. Through nine developed com-pounds one of them get approved by the FDA. Animal models and in vitro models are both currently used to investigate pharma-codynamics and pharmacokinetics of new drugs. in vitro models are promising tools to predict drug toxicity and drug screening tests in pharmaceutical industry. In liver, isolated primary hu-man hepatocytes (PHH) are the gold standard for drug screen-ing approaches which representing the metabolizing profile like in vivo environments. However, these cells lose their functional metabolic features 72 hours after cultivation. In recent years some attempts have been done to extend the maintenance and functionality of PHH, including 3D cultures, micro-fluid flow cultures, 3D printing, and using synthetic and natural materials. Supportive cells in 3D culture are reliable way to improve main-tanance of PHH through increase cell-cell interaction and growth factors secretion. In this work we used mesenchymal stromal cells (MSCs), human umbilical vein endothelial cells (HUVECs) as supportive cells and liver matrix derived microparticles (MPs) as natural extra cellular matrix (ECM).Materials and Methods: Three cell types including PHH, MSCs, and HUVECs co-cultured in AgreewellTM plate and or-ganoids harvested the day after. The MPs were fabricated with a water in oil method. The organoids were cultured in Aggrewell plate in 2 groups with and without MPs.Results: MPs size distribution measured, that was approximate-ly between 8-12 um. The MPs were able to incorporation into organoids successfully. The ratio of incorporation was > 80%. The toxicity assay examined, data was shown that the MPs have no toxicity for cells. The ratio of MPs to cells set up in 1:2 ratio. Conclusion: Organoids are reliable tool to investigation of drug screening and toxicity assays in liver.

کلیدواژه ها:

Drug Screening ، Microparticles ، Organoid ، Hepato-cyte