000089606 001__ 89606 000089606 005__ 20210902121636.0 000089606 0247_ $$2doi$$a10.1002/hep.31037 000089606 0248_ $$2sideral$$a117653 000089606 037__ $$aART-2020-117653 000089606 041__ $$aeng 000089606 100__ $$aSchneeberger, K. 000089606 245__ $$aLarge-scale production of LGR5-positive bipotential human liver stem cells 000089606 260__ $$c2020 000089606 5060_ $$aAccess copy available to the general public$$fUnrestricted 000089606 5203_ $$aBackground and Aims: The gap between patients on transplant waiting lists and available donor organs is steadily increasing. Human organoids derived from leucine‐rich repeat‐containing G protein‐coupled receptor 5 (LGR5)–positive adult stem cells represent an exciting new cell source for liver regeneration; however, culturing large numbers of organoids with current protocols is tedious and the level of hepatic differentiation is limited. Approach and Results: Here, we established a method for the expansion of large quantities of human liver organoids in spinner flasks. Due to improved oxygenation in the spinner flasks, organoids rapidly proliferated and reached an average 40‐fold cell expansion after 2 weeks, compared with 6‐fold expansion in static cultures. The organoids repopulated decellularized liver discs and formed liver‐like tissue. After differentiation in spinner flasks, mature hepatocyte markers were highly up‐regulated compared with static organoid cultures, and cytochrome p450 activity reached levels equivalent to hepatocytes. Conclusions: We established a highly efficient method for culturing large numbers of LGR5‐positive stem cells in the form of organoids, which paves the way for the application of organoids for tissue engineering and liver transplantation. 000089606 536__ $$9info:eu-repo/grantAgreement/ES/ISCIII/FIS/PI18-00529$$9info:eu-repo/grantAgreement/ES/ISCIII/FIS/PI15-00563$$9This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 874586-ORGANTRANS$$9info:eu-repo/grantAgreement/EC/H2020/874586/EU/Organ 3D printing for liver regenerative medicine/ORGANTRANS 000089606 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc-sa$$uhttp://creativecommons.org/licenses/by-nc-sa/3.0/es/ 000089606 590__ $$a17.425$$b2020 000089606 591__ $$aGASTROENTEROLOGY & HEPATOLOGY$$b6 / 92 = 0.065$$c2020$$dQ1$$eT1 000089606 592__ $$a5.488$$b2020 000089606 593__ $$aMedicine (miscellaneous)$$c2020$$dQ1 000089606 593__ $$aHepatology$$c2020$$dQ1 000089606 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion 000089606 700__ $$aSánchez-Romero, N. 000089606 700__ $$aYe, S.C. 000089606 700__ $$avan Steenbeek, F.G. 000089606 700__ $$aOosterhoff, L.A. 000089606 700__ $$aPla Palacin, I. 000089606 700__ $$aChen, C. 000089606 700__ $$avan Wolferen, M.E. 000089606 700__ $$avan Tienderen, G. 000089606 700__ $$aLieshout, R. 000089606 700__ $$aColemonts-Vroninks, H. 000089606 700__ $$aSchene, I. 000089606 700__ $$aHoekstra, R. 000089606 700__ $$aVerstegen, M.M.A. 000089606 700__ $$avan der Laan, L.J.W. 000089606 700__ $$aPenning, L.C. 000089606 700__ $$aFuchs, S.A. 000089606 700__ $$aClevers, H. 000089606 700__ $$aDe Kock, J. 000089606 700__ $$aBaptista, P.M. 000089606 700__ $$aSpee, B. 000089606 773__ $$g72, 1 (2020), 257-270$$pHepatology$$tHEPATOLOGY$$x0270-9139 000089606 8564_ $$s533782$$uhttps://zaguan.unizar.es/record/89606/files/texto_completo.pdf$$yVersión publicada 000089606 8564_ $$s36007$$uhttps://zaguan.unizar.es/record/89606/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada 000089606 909CO $$ooai:zaguan.unizar.es:89606$$particulos$$pdriver 000089606 951__ $$a2021-09-02-08:56:05 000089606 980__ $$aARTICLE