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Biofabrication of size-controlled liver microtissues incorporated with ECM-derived microparticles to prolong hepatocyte function

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Bio-Design and Manufacturing 2021年第4期4卷 790-805页

作者： Zahra Heydari Ibrahim Zarkesh Mohammad-Hossein Ghanian Mahdokht HAghdaei Svetlana Kotova Ensieh Zahmatkesh Zahra Farzaneh Abbas Piryaei Iman Akbarzadeh Anastasia Shpichka Roberto Gramignoli Peter Timashev Hossein Baharvand Massoud Vosough Department of Developmental Biology University of Science and CultureACECR14155-4364 TehranIran Department of Regenerative Medicine Cell Science Research CentreRoyan Institute for Stem Cell Biology and TechnologyACECR14155-4364 TehranIran Department of Stem Cells and Developmental Biology Cell Science Research CenterRoyan Institute for Stem Cell Biology and TechnologyACECR14155-4364 TehranIran Department of Cell Engineering Cell Science Research CenterRoyan Institute for Stem Cell Biology and TechnologyACECR14155-4364 TehranIran Transplant Research Center Shiraz University of Medical Sciences71936-13311 ShirazIran N.N.Semenov Federal Research Center for Chemical Physics Russian Academy of SciencesMoscow 117977Russia Department of Biology and Anatomical Sciences School of MedicineShahid Beheshti University of Medical Sciences14155-4364 TehranIran Department of Tissue Engineering and Applied Cell Sciences School of Advanced Technologies in MedicineShahid Beheshti University of Medical Sciences14155-4364 TehranIran Institute for Regenerative Medicine Sechenov UniversityMoscow 117977Russia Division of Pathology Department of Laboratory MedicineKarolinska Institute17177 StockholmSweden

Multicellular microtissues of primary human hepatocytes(PHHs)co-cultured with other supporting cell types are a promis-ing model for drug screening and toxicological ***,these liver microtissues(LMs)rapidly lose their functions during ex vivo ***,in order to mimic the cellular and structural hepatic microenvironment,we co-cultured PHHs with human mesenchymal stromal cells(MSCs)and human umbilical vein endothelial cells(HUVECs)in the presence of cell-sized microparticles(MPs)derived from liver extracellular matrix(LEMPs).The microwell culture platform enabled biofabrication of size-controlled multicellular microtissues(PHH:HUVEC:MSC=3:2:1)with efficient LEMP incorporation(about 70%at a 2:1 ratio of cells:MP).The biofabricated liver microtissues(BLMs)were cultured ex vivo for 14 days and compared to the cell-only LM in terms of gene and protein expression,functional activity,cytochrome P450(CYP450)enzyme inducibility,and drug *** results supported superior hepatic-related gene expression,functional activity,and polarity for PHH in BLM compared to ***450 enzyme inducibility and dose-responsive sensitivity to toxic drugs were significantly higher in the BLM *** conclusion,microtissue engineering by incorporation of tissue-specific microparticles within a multicellular microtissue can offer some advantages for drug discovery studies and cell transplantation *** the near future,this approach could generate a scalable platform of several functional biofabricated microtissues representing different organs.

关键词： liver microtissue Hepatocyte maintenance Microparticles Decellularized tissue Drug screening

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