Nascent matrix deposition supports alveolar organoid formation from aggregates in synthetic hydrogels

Madeline K Eiken; Charlie J Childs; Lindy K Brastrom; Tristan Frum; Eleanor M Plaster; Donia W Ahmed; Ryan C Spencer; Orren Shachaf; Suzanne Pfeiffer; Justin E Levine; Konstantinos-Dionysios Alysandratos; Darrell N Kotton; Jason R Spence; Claudia Loebel

doi:10.1016/j.stemcr.2024.11.006

Nascent matrix deposition supports alveolar organoid formation from aggregates in synthetic hydrogels

Stem Cell Reports. 2024 Dec 12:102376. doi: 10.1016/j.stemcr.2024.11.006. Online ahead of print.

Affiliations

¹ Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA.
² Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI, USA.
³ Department of Biomedical Engineering, University of Texas, Austin, TX, USA.
⁴ Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA 02118, USA; The Pulmonary Center and Department of Medicine, Boston University Chobanian & Avedisian School of Medicine, Boston, MA 02118, USA.
⁵ Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA; Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI, USA; Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI, USA; Program in Cell and Molecular Biology, University of Michigan Medical School, Ann Arbor, MI, USA.
⁶ Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA; Department of Materials Science and Engineering, University of Michigan, Ann Arbor, MI, USA. Electronic address: [email protected].

PMID: 39672155
DOI: 10.1016/j.stemcr.2024.11.006

Abstract

Human induced pluripotent stem cell (iPSC)-derived alveolar organoids have emerged as a system to model the alveolar epithelium in homeostasis and disease. However, alveolar organoids are typically grown in Matrigel, a mouse sarcoma-derived basement membrane matrix that offers poor control over matrix properties, prompting the development of synthetic hydrogels as a Matrigel alternative. Here, we develop a two-step culture method that involves pre-aggregation of organoids in hydrogel-based microwells followed by embedding in a synthetic hydrogel that supports alveolar organoid growth, while also offering considerable control over organoid and hydrogel properties. We find that the aggregated organoids secrete their own nascent extracellular matrix (ECM) both in the microwells and upon embedding in synthetic hydrogels, which supports their growth. Thus, the synthetic hydrogels described here allow us to de-couple exogenous and nascent ECM to interrogate the role of ECM in organoid formation.

Keywords: alveolospheres; extracellular matrix; hydrogels; lung; organoids.