The Microfluidic Environment Reveals a Hidden Role of Self-Organizing Extracellular Matrix in Hepatic Commitment and Organoid Formation of hiPSCs
Federica Michielin,
Giovanni G. Giobbe,
Camilla Luni,
Qianjiang Hu,
Ida Maroni,
Michael R. Orford,
Anna Manfredi,
Lucio Di Filippo,
Anna L. David,
Davide Cacchiarelli,
Paolo De Coppi,
Simon Eaton,
Nicola Elvassore
Affiliations
Federica Michielin
Great Ormond Street Institute of Child Health, University College London, WC1N1EH London, UK
Giovanni G. Giobbe
Great Ormond Street Institute of Child Health, University College London, WC1N1EH London, UK
Camilla Luni
Shanghai Institute for Advanced Immunochemical Studies (SIAIS), ShanghaiTech University, 201210 Shanghai, China
Qianjiang Hu
Shanghai Institute for Advanced Immunochemical Studies (SIAIS), ShanghaiTech University, 201210 Shanghai, China
Ida Maroni
Department of Industrial Engineering, University of Padova, 35131 Padova, Italy; Venetian Institute of Molecular Medicine (VIMM), 35129 Padova, Italy
Michael R. Orford
Great Ormond Street Institute of Child Health, University College London, WC1N1EH London, UK
Anna Manfredi
Telethon Institute of Genetics and Medicine (TIGEM), Armenise/Harvard Laboratory of Integrative Genomics, 80078 Pozzuoli, Italy
Lucio Di Filippo
Next Generation Diagnostic Srl, 80078 Pozzuoli, Italy
Anna L. David
Elizabeth Garrett Anderson Institute for Women’s Health, University College London, WC1E 6AU London, UK
Davide Cacchiarelli
Telethon Institute of Genetics and Medicine (TIGEM), Armenise/Harvard Laboratory of Integrative Genomics, 80078 Pozzuoli, Italy; Department of Translational Medicine, University of Naples “Federico II,” 80131 Naples, Italy
Paolo De Coppi
Great Ormond Street Institute of Child Health, University College London, WC1N1EH London, UK; Specialist Neonatal and Paediatric Surgery, Great Ormond Street Hospital, WC1N 3JH London, UK
Simon Eaton
Great Ormond Street Institute of Child Health, University College London, WC1N1EH London, UK
Nicola Elvassore
Great Ormond Street Institute of Child Health, University College London, WC1N1EH London, UK; Shanghai Institute for Advanced Immunochemical Studies (SIAIS), ShanghaiTech University, 201210 Shanghai, China; Department of Industrial Engineering, University of Padova, 35131 Padova, Italy; Venetian Institute of Molecular Medicine (VIMM), 35129 Padova, Italy; Corresponding author
Summary: The specification of the hepatic identity during human liver development is strictly controlled by extrinsic signals, yet it is still not clear how cells respond to these exogenous signals by activating secretory cascades, which are extremely relevant, especially in 3D self-organizing systems. Here, we investigate how the proteins secreted by human pluripotent stem cells (hPSCs) in response to developmental exogenous signals affect the progression from endoderm to the hepatic lineage, including their competence to generate nascent hepatic organoids. By using microfluidic confined environment and stable isotope labeling with amino acids in cell culture-coupled mass spectrometry (SILAC-MS) quantitative proteomic analysis, we find high abundancy of extracellular matrix (ECM)-associated proteins. Hepatic progenitor cells either derived in microfluidics or exposed to exogenous ECM stimuli show a significantly higher potential of forming hepatic organoids that can be rapidly expanded for several passages and further differentiated into functional hepatocytes. These results prove an additional control over the efficiency of hepatic organoid formation and differentiation for downstream applications.
