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JESSE BROWN VA MEDICAL CENTER

3 ARTICLES PUBLISHED IN JoVE

Bioengineering

Epithelial Cell Repopulation and Preparation of Rodent Extracellular Matrix Scaffolds for Renal Tissue Development

Joseph S. Uzarski^1,2, Jimmy Su^1,2,3,4, Yan Xie^1,2, Zheng J. Zhang^1,2, Heather H. Ward⁵, Angela Wandinger-Ness⁶, William M. Miller^7,8, Jason A. Wertheim^1,2,3,4,8,9

¹Comprehensive Transplant Center, Feinberg School of Medicine, Northwestern University, ²Department of Surgery, Feinberg School of Medicine, Northwestern University, ³Department of Biomedical Engineering, Northwestern University, ⁴Simpson Querrey Institute for BioNanotechnology in Medicine, Northwestern University, ⁵Department of Internal Medicine, University of New Mexico HSC, ⁶Department of Pathology, University of New Mexico HSC, ⁷Department of Chemical and Biological Engineering, Northwestern University, ⁸Chemistry of Life Processes Institute, Northwestern University, ⁹Department of Surgery, Jesse Brown VA Medical Center

This protocol describes decellularization of Sprague Dawley rat kidneys by antegrade perfusion of detergents through the vasculature, producing acellular renal extracellular matrices that serve as templates for repopulation with human renal epithelial cells. Recellularization and use of the resazurin perfusion assay to monitor growth is performed within specially-designed perfusion bioreactors.

Biology

Methods to Study Epithelial Transport Protein Function and Expression in Native Intestine and Caco-2 Cells Grown in 3D

Arivarasu N. Anabazhagan^*1, Ishita Chatterjee^*1, Shubha Priyamvada¹, Anoop Kumar¹, Sangeeta Tyagi¹, Seema Saksena^1,2, Waddah A. Alrefai^1,2, Pradeep K. Dudeja^1,2, Ravinder K. Gill¹

¹Department of Medicine, University of Illinois at Chicago, ²Department of Research, Jesse Brown VA Medical Center

We describe simple methods to study the regulation of intestinal serotonin transporter (SERT) function and expression using an in vitro cell culture model of Caco-2 cells grown in 3D and an ex vivo model of mouse intestines. These methods are applicable to the study of other epithelial transporters.

Kidney Organoid Generation at the Air-Liquid Interface

Ashwani Kumar Gupta^1,2, David Z. Ivancic¹, Bilal A. Naved^1,3, Jason A. Wertheim^1,2,3,4,5,6, Leif Oxburgh⁷

¹Comprehensive Transplant Center, Northwestern University Feinberg School of Medicine, ²Department of Surgery, Northwestern University Feinberg School of Medicine, ³Department of Biomedical Engineering, McCormick School of Engineering, Northwestern University, ⁴Simpson Querrey Institute for BioNanotechnology, Northwestern University, ⁵Chemistry of Life Processes Institute, Northwestern University, ⁶Department of Surgery, Jesse Brown VA Medical Center, ⁷The Rogosin Institute

This protocol describes asynchronous mixing of human embryonic stem cells derived kidney progenitors at the air-liquid interface to efficiently generate kidney organoids.