title: Functional ultrasound imaging for assessment of extracellular matrix scaffolds used for liver organoid formation.
abstract: A method of 3D functional ultrasound imaging has been developed to enable non-destructive assessment of extracellular matrix scaffolds that have been prepared by decellularization protocols and are intended for recellularization to create organoids. A major challenge in organ decellularization is retaining patent micro-vascular structures crucial for nutrient access and functionality of organoids. The imaging method described here provides statistical distributions of flow rates throughout the tissue volumes, 3D vessel network architecture visualization, characterization of microvessel volumes and sizes, and delineation of matrix from vascular circuits. The imaging protocol was tested on matrix scaffolds that are tissue-specific, but not species-specific, matrix extracts, prepared by a process that preserved >98% of the collagens, collagen-associated matrix components, and matrix-bound growth factors and cytokines. Image-derived data are discussed with respect to assessment of scaffolds followed by proof-of-concept studies in organoid establishment using Hep3B, a human hepatoblast-like cell line. Histology showed that the cells attached to scaffolds with patent vasculature within minutes, achieved engraftment at near 100%, expressed liver-specific functions within 24 h, and yielded evidence of proliferation and increasing differentiation of cells throughout the two weeks of culture studies. This imaging method should prove valuable in analyses of such matrix scaffolds.
authors.affiliation: UNC/NCSU Joint Department of Biomedical Engineering, UNC School of Medicine, Chapel Hill, NC 27599, USA. Electronic address: gessner.ryan@gmail.com.
authors.forename: Ryan C
authors.initials: RC
authors.lastname: Gessner
authors.forename: Ariel D
authors.initials: AD
authors.lastname: Hanson
authors.forename: Steven
authors.initials: S
authors.lastname: Feingold
authors.forename: Avery T
authors.initials: AT
authors.lastname: Cashion
authors.forename: Ana
authors.initials: A
authors.lastname: Corcimaru
authors.forename: Bryant T
authors.initials: BT
authors.lastname: Wu
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Mullins
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Aylward
authors.forename: Lola M
authors.initials: LM
authors.lastname: Reid
authors.forename: Paul A
authors.initials: PA
authors.lastname: Dayton
mesh_terms: D000818:Animals; D002460:Cell Line; D005109:Extracellular Matrix / Q000737:chemistry / Q000000981:diagnostic imaging* / Q000648:ultrastructure; D006801:Humans; D008099:Liver / Q000166:cytology / Q000000981:diagnostic imaging* / Q000648:ultrastructure; D009940:Organoids / Q000166:cytology*; D051381:Rats; D017208:Rats, Wistar; D054457:Tissue Scaffolds / Q000737:chemistry*; D014463:Ultrasonography
keywords: Acoustic angiography; Extracellular matrix; Liver; Microbubble; Organoids; Scaffolds