Hormonally and chemically defined expansion conditions for organoids of biliary tree Stem Cells  

作　　者：Wencheng Zhang Yangyang Cui Mengqi Lu Mingyang Xu Yuting Li Haimeng Song Yi Luo Jinjia Song Yong Yang Xicheng Wang Lijun Liao Yunfang Wang Lola Reid Zhiying He 

机构地区：[1]Institute for Regenerative Medicine,Medical Innovation Center and State Key Laboratory of Cardiology,Shanghai East Hospital,School of Life Sciences and Technology,Tongji University,Shanghai,200123,China [2]Shanghai Engineering Research Center of Stem Cells Translational Medicine,Shanghai,200335,China [3]Shanghai Institute of Stem Cell Research and Clinical Translation,Shanghai,200120,China [4]Postgraduate Training Base of Shanghai East Hospital,Jinzhou Medical University,Jinzhou,Liaoning,121001,China [5]Department of General Surgery,The First Affiliated Hospital of Nanchang University,Nanchang,Jiangxi,330006,China [6]Department of Anesthesiology and Pain Management,Shanghai East Hospital,Tongji University School of Medicine,Shanghai,200123,China [7]Hepatobiliary and Pancreatic Center,Medical Research Center,Beijing Tsinghua Changgung Hospital,Beijing,102218,China [8]Departments of Cell Biology and Physiology,Program in Molecular Biology and Biotechnology,UNC School of Medicine,Chapel Hill,NC,27599,United States

出　　处：《Bioactive Materials》2024年第11期672-695,共24页生物活性材料（英文）

基　　金：Major Program of National Key Research and Development Project(2020YFA0112600,2019YFA0801502);National Natural Science Foundation of China(82173019,82270638,8220374,82300718);Project of Shanghai Science and Technology Commission(22ZR1451100,22Y11908500);Peak Disciplines(Type IV)of Institutions of Higher Learning in Shanghai,and Shanghai Engineering Research Center of Stem Cells Translational Medicine(20DZ2255100).

摘　　要：Wholly defined ex vivo expansion conditions for biliary tree stem cell(BTSC)organoids were established,consisting of a defined proliferative medium(DPM)used in combination with soft hyaluronan hydrogels.The DPM consisted of commercially available Kubota's Medium(KM),to which a set of small molecules,particular paracrine signals,and heparan sulfate(HS)were added.The small molecules used were DNA methyltransferase inhibitor(RG108),TGF-βType I receptor inhibitor(A83-01),adenylate cyclase activator(Forskolin),and L-type Ca2+channel agonist(Bay K8644).A key paracrine signal proved to be R-spondin 1(RSPO1),a secreted protein that activates Wnts.Soluble hyaluronans,0.05%sodium hyaluronate,were used with DPM to expand monolayer cultures.Expansion of organoids was achieved by using DPM in combination with embedding organoids in Matrigel that was replaced with a defined thiol-hyaluronan triggered with PEGDA to form a hydrogel with a rheology[G*]of less than 100 Pa.The combination is called the BTSC-Expansion-Glycogel-System(BEX-gel system)for expanding BTSCs as a monolayer or as organoids.The BTSC organoids were expanded more than 3000-fold ex vivo in the BEX-gel system within 70 days while maintaining phenotypic traits indicative of stem/progenitors.Stem-cell-patch grafting of expanded BTSC organoids was performed on the livers of Fah-/-mice with tyrosinemia and resulted in the rescue of the mice and restoration of their normal liver functions.The BEX-gel system for BTSC organoid expansion provides a strategy to generate sufficient numbers of organoids for the therapeutic treatments of liver diseases.

关 键 词：Biliary tree stem cells(BTSCs) ORGANOIDS Paracrine signals Defined Proliferative Medium(DPM) BTSC-Expansion-Glycogel(BEX-gel) 

分 类 号：O62[理学—有机化学]