August 2015 - This Month in JoVE: Isolating Stem Cells, Bioengineering the Kidney, and Getting Kids to eat Carrots

Protocol

Generation of Integration-free Human Induced Pluripotent Stem Cells Using Hair-derived Keratinocytes

Sandy S.C. Hung, Alice Pébay, Raymond C.B. Wong

Centre for Eye Research Australia & Department of Ophthalmology, University of Melbourne

This manuscript provides a step-by-step procedure for the derivation and maintenance of human keratinocytes from plucked hair and subsequent generation of integration-free human induced pluripotent stem cells (hiPSCs) by episomal vectors.

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 Jenny 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.

A Treatment Package without Escape Extinction To Address Food Selectivity

Jessica Weber¹, Anibal Gutierrez, Jr.²

¹Florida International University, ²Department of Psychology and the Center for Children and Families, Florida International University

Feeding difficulties are a common problem for children with developmental disorders, including autism, and behavioral interventions often include escape extinction. Recent research has begun to evaluate treatments that do not include escape extinction. This manuscript describes a multicomponent treatment package that does not use escape extinction to treat feeding difficulties.

A Novel Culture Model for Human Pluripotent Stem Cell Propagation on Gelatin in Placenta-conditioned Media

Ji-Hye Jung, Byung Soo Kim

Department of Biomedical Science, Graduate School of Medicine, Korea University

This protocol provides a simple and efficient way to propagate human pluripotent stem cells (hPSCs) using only conditioned media derived from the human placenta in a gelatin-coated dish without additional exogenous supplementation or hPSC-specific synthetic substrata.