U.S. Army Institute of Surgical Research View Institution's Website 5 articles published in JoVE Developmental Biology Production and Administration of Therapeutic Mesenchymal Stem/Stromal Cell (MSC) Spheroids Primed in 3-D Cultures Under Xeno-free Conditions Joni H. Ylostalo1, Nikolay Bazhanov2, Arezoo Mohammadipoor3, Thomas J. Bartosh4 1Department of Biology, University of Mary Hardin-Baylor, 2Department of Pediatrics, University of Texas Medical Branch, 3Multi-Organ Support Technology Task Area, U.S. Army Institute of Surgical Research, 4Internal Medicine, Texas A&M University Health Science Center The therapeutic potential of mesenchymal stem/stromal cells (MSCs) is well-documented, however the best method of preparing the cells for patients remains controversial. Herein, we communicate protocols to efficiently generate and administer therapeutic spherical aggregates or 'spheroids' of MSCs primed under xeno-free conditions for experimental and clinical applications. Biology Physiology Lab Demonstration: Glomerular Filtration Rate in a Rat Carmen Hinojosa-Laborde1, Brian Jespersen2, Robert Shade3 1Tactical Combat Casualty Care Research, U.S. Army Institute of Surgical Research, 2Department of Pharmacology and Toxicology, Michigan State University, 3Southwest National Primate Research Center, Texas Biomedical Research Institute The purpose of this protocol is to demonstrate the principles and techniques for measuring and calculating glomerular filtration rate, urine flow rate, and excretion of sodium and potassium in a rat. This demonstration can be used to provide students with an overall conceptual understanding of how to measure renal function. Developmental Biology Deriving Retinal Pigment Epithelium (RPE) from Induced Pluripotent Stem (iPS) Cells by Different Sizes of Embryoid Bodies Alberto Muñiz1, Kaini R. Ramesh1, Whitney A. Greene1, Jae-Hyek Choi1, Heuy-Ching Wang1 1Ocular Trauma, U.S. Army Institute of Surgical Research The objective of this report is to describe the protocols to derive the retinal pigment epithelium (RPE) from induced pluripotent stem (iPS) cells using different sizes of embryoid bodies. Bioengineering Constructing a Collagen Hydrogel for the Delivery of Stem Cell-loaded Chitosan Microspheres David O. Zamora*1, Shanmugasundaram Natesan*1, Robert J. Christy1 1Department of Regenerative Medicine, United States Army Institute of Surgical Research A major hurdle in current stem cell therapies is determining the most effective method to deliver these cells to host tissues. Here, we describe a chitosan-based delivery method that is efficient and simple in approach, while allowing adipose-derived stem cells to maintain their multipotency. Bioengineering Engineering a Bilayered Hydrogel to Control ASC Differentiation Shanmugasundaram Natesan1, David O. Zamora1, Laura J. Suggs2, Robert J. Christy1 1Department of Extremity Trauma Research and Regenerative Medicine, United States Army Institute of Surgical Research, 2Department of Biomedical Engineering, The University of Texas at Austin This protocol focuses on utilizing the inherent ability of stem cells to take cue from their surrounding extracellular matrix and be induced to differentiate into multiple phenotypes. This methods manuscript extends our description and characterization of a model utilizing a bilayered hydrogel, composed of PEG-fibrin and collagen, to simultaneously co-differentiate adipose-derived stem cells1.