biostudies-literatureGong SKey R&D Program of ZhejiangNational Natural Science Foundation of ChinaNational Key Research and Development Program of Chinae70033https://www.ebi.ac.uk/biostudies/studies/S-EPMC12414634biostudies-literatureUnknown58(9)Liver transplantation is currently the sole definitive treatment option for end-stage liver failure. However, a significant shortage of donors prevails due to high clinical demands. Recently, human liver organoids have shown significant potential in regenerative medicine for liver diseases. Nevertheless, current static cultures of organoids grown in well-plates heavily rely on extracellular matrix hydrogels (Matrigel), thereby limiting both the scalability and quantity of organoid culture. In this study, we present a groundbreaking culture mode that eliminates all reliance on extracellular matrix hydrogels, enabling the successful preparation of functional human liver ductal organoids (LDOs) based on the cell suspension culture mode in a mechanically stirred bioreactor. Initially, the deveCell proliferationScalable Matrigel-Free Suspension Culture for Generating High-Quality Human Liver Ductal Organoids.PMC124146342023YFC230680092159202824706902024C031492023C03046Shen MGong SYang LYan QHe JZheng SAshraf KHe KRehman TULi WLiu YGuo MYang ZLuo XMohsin AfalseScalable Matrigel-Free Suspension Culture for Generating High-Quality Human Liver Ductal Organoids.Liver transplantation is currently the sole definitive treatment option for end-stage liver failure. However, a significant shortage of donors prevails due to high clinical demands. Recently, human liver organoids have shown significant potential in regenerative medicine for liver diseases. Nevertheless, current static cultures of organoids grown in well-plates heavily rely on extracellular matrix hydrogels (Matrigel), thereby limiting both the scalability and quantity of organoid culture. In this study, we present a groundbreaking culture mode that eliminates all reliance on extracellular matrix hydrogels, enabling the successful preparation of functional human liver ductal organoids (LDOs) based on the cell suspension culture mode in a mechanically stirred bioreactor. Initially, the deve2025-01-01T00:00:00Z2025 Sep2026-06-02T22:36:53.614Z2026-05-28T03:06:57.57ZS-EPMC124146344016695510.1111/cpr.70033