Comprehensive Characterization of Solubilized Lung ECM

Non-diseased, non-smoking human lung donor tissue was obtained (International Institute for the Advancement of Medicine), decellularized, and solubilized following protocols previously published by our group^{31 (link),33 (link),70 (link)}. The solubilized lung ECM (sECM) was aliquoted and stored at -80 °C until further use. The ECM was diluted for colorimetric assays to measure collagen, elastin, sulphated glycosaminoglycan, and hyaluronan content (Biocolor, United Kingdom). Fibronectin and laminin were quantified using the Human Laminin ELISA kit (Abcam, United Kingdom) and Human Fibronectin ELISA Kit (R&D Systems, Minneapolis, MN), respectively. To estimate spectral counts of specific collagen types and the composition of growth factors, the sECM was sent to and processed by RayBiotech (Peachtree Corners, GA) using the Human L3 Glass Slide Array and Human Growth Factor Array Q1. Additionally, total protein content of the sECM was quantified with a Pierce™ BCA Protein Assay Kit (Thermofisher, Waltham, MA). A total of six individual lung donors were decellularized, solubilized, and analyzed for the solubilized ECM content.

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Leach T., Gandhi U., Reeves K.D., Stumpf K., Okuda K., Marini F.C., Walker S.J., Boucher R., Chan J., Cox L.A., Atala A, & Murphy S.V. (2023). Development of a novel air–liquid interface airway tissue equivalent model for in vitro respiratory modeling studies. Scientific Reports, 13, 10137.

Publication 2023

Human Laminin ELISA kit Human Fibronectin ELISA Kit Human Growth Factor Array Q1 Pierce™ BCA Protein Assay Kit

Assay Collagen Colorimetric Donor tissue Donors individual Ecm protein Elastin Elisa Fibronectin Glycosaminoglycan Growth factor Human Hyaluronan Laminin Lung Protein a

Corresponding Organization :

Other organizations : Wake Forest University, Forest Institute, Virginia Tech - Wake Forest University School of Biomedical Engineering & Sciences, University of North Carolina at Chapel Hill, Lung Institute

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Collagen content
Elastin content
Sulphated glycosaminoglycan content
Hyaluronan content
Fibronectin content
Laminin content
Specific collagen types (via spectral counts)
Composition of growth factors
Total protein content

control variables

Non-diseased, non-smoking human lung donor tissue

controls

No positive or negative controls were explicitly mentioned in the protocol.

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