Generation of 2.5D lung bud organoids from human induced pluripotent stem cells

Issue title: 40th Conference of the German Society for Clinical Microcirculation and Hemorheology, 5-6 November 2021, Senftenberg, Germany

Guest editors: J.-H. Küpper, A. Krüger-Genge and F. Jung

Article type: Research Article

Authors: Xu, Xun^{a; 1} | Nie, Yan^{a; b; 1} | Wang, Weiwei^{a; 1} | Ullah, Imran^a | Tung, Wing Tai^{a; b} | Ma, Nan^{a; c; *} | Lendlein, Andreas^{a; b; c; *}

Affiliations: [a] Institute of Active Polymers and Berlin-Brandenburg Centre for Regenerative Therapies, Helmholtz-Zentrum Hereon, Teltow, Germany | [b] Institute of Biochemistry and Biology, University of Potsdam, Potsdam, Germany | [c] Institute of Chemistry and Biochemistry, Free University of Berlin, Berlin, Germany

Correspondence: [*] Corresponding author: Prof. Dr. Nan Ma, and Prof. Dr. Andreas Lendlein, Institute of Active Polymers, Helmholtz-Zentrum Hereon, Kantstr. 55, 14513 Teltow, Germany. E-mail: nan.ma@hereon.de; andreas.lendlein@hereon.de.

Note: [1] These authors contributed equally to this work.

Abstract: Human induced pluripotent stem cells (hiPSCs) are a promising cell source to generate the patient-specific lung organoid given their superior differentiation potential. However, the current 3D cell culture approach is tedious and time-consuming with a low success rate and high batch-to-batch variability. Here, we explored the establishment of lung bud organoids by systematically adjusting the initial confluence levels and homogeneity of cell distribution. The efficiency of single cell seeding and clump seeding was compared. Instead of the traditional 3D culture, we established a 2.5D organoid culture to enable the direct monitoring of the internal structure via microscopy. It was found that the cell confluence and distribution prior to induction were two key parameters, which strongly affected hiPSC differentiation trajectories. Lung bud organoids with positive expression of NKX 2.1, in a single-cell seeding group with homogeneously distributed hiPSCs at 70% confluence (SC_70%_hom) or a clump seeding group with heterogeneously distributed cells at 90% confluence (CL_90%_het), can be observed as early as 9 days post induction. These results suggest that a successful lung bud organoid formation with single-cell seeding of hiPSCs requires a moderate confluence and homogeneous distribution of cells, while high confluence would be a prominent factor to promote the lung organoid formation when seeding hiPSCs as clumps. 2.5D organoids generated with defined culture conditions could become a simple, efficient, and valuable tool facilitating drug screening, disease modeling and personalized medicine.

Keywords: Lung organoid, human induced pluripotent stem cell, cell culture

DOI: 10.3233/CH-219111

Journal: Clinical Hemorheology and Microcirculation, vol. 79, no. 1, pp. 217-230, 2021