1Materials Science and Engineering, CSIRO
We describe the use of the monoclonal antibodies TG30 (CD9) and GCTM-2 for the combined detection of cell surface antigens via fluorescence activated cell sorting (FACS) for the identification and enrichment of live human embryonic stem cells (hESC) using positive selection and also the use of negative selection to purge hESCs from a mixed cell population.
1School of Molecular Biosciences, Washington State University, 2Center for Reproductive Biology, Washington State University
<em>Caenorhabditis elegan</em> is a useful model to explore the functions of polyunsaturated fatty acids in development and physiology. This protocol describes an efficient method of supplementing the <em>C. elegans</em> diet with polyunsaturated fatty acids.
1Department of Craniofacial Development and Stem Cell Biology, and Department of Orthodontics, Dental Institute, Guy's Hospital, UK, King's College London, 2Department of Pediatric Dentistry and Orthodontics, College of Dentistry, King Saud University, Kingdom of Saudi Arabia
Here we detail a method to culture tooth germs in mandible slices using a tissue chopper. This method allows unique access to the tooth during development, providing excellent opportunity for manipulation and lineage tracing, not available using more traditional culture methods.
1Department of Biological Sciences, Ohio University
"Freeze-cracking," a method for exposing the inner tissues of the nematode C. elegans to antibodies for protein localization, is demonstrated.
JoVE Clinical and Translational Medicine
1Division of Gastroenterology, Case Western Reserve University School of Medicine, Cleveland, 2Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, 3Digestive Health Research Center, Case Western Reserve University School of Medicine, Cleveland
Over the past few years, new generation endoscopes have emerged as important diagnostic research aids for evaluating murine colitis and colorectal tumors. We present herein a detailed protocol for endoscopic assessment of inflammation and colorectal tumors in mice, as well as a novel scoring system that uses decimal identifiers to document the endoscopic severity of colitis and colorectal tumors.
1Cytori Therapeutics Inc, 2Division of Cardiac Surgery, Department of Surgery, David Geffen School of Medicine at UCLA, 3Division of Plastic and Reconstructive Surgery, Department of Surgery, David Geffen School of Medicine at UCLA, 4Department of Orthopedic Surgery, David Geffen School of Medicine at UCLA, 5Regenerative Bioengineering and Repair Laboratory, David Geffen School of Medicine at UCLA
In 2001, researchers at UCLA described the isolation of a population of adult stem cells, termed Adipose-derived Stem Cells or ASCs, from adipose tissue. This article outlines the isolation of ASCs from lipoaspirates using a manual, enzymatic digestion protocol using collagenase.
1Department of Surgery, Weill Cornell Medical College
A simple method to derive and maintain spermatogonial stem and progenitor cell lines from adult mice is presented here. The method utilizes feeder cells originating from the somatic cell compartment of the adult mouse testis. This technique is applicable to common mouse strains, including transgenic, knock-out, and knock-in mice.
?Department of Biological Sciences, University of Toledo
Imaging of centrosomal proteins during Drosophila spermatogenesis is a powerful method to identify new proteins critical for centrosome biology as well as to elucidate the particular function of known players in this process.
1Humanitas Clinical and Research Center, Italy, 2Institute of Genetic and Biomedical Research (IRGB), National Research Council (CNR)
Pluripotent stem cells, either embryonic or induced pluripotent stem (iPS) cells, constitute a valuable source of human differentiated cells, including cardiomyocytes. Here, we will focus on cardiac induction of iPS cells, showing how to use them to obtain functional human cardiomyocytes through an embryoid bodies-based protocol.
JoVE Immunology and Infection
1School of Chemistry, Food and Pharmacy, The University of Reading, 2Department of Nutritional Sciences, The University of Reading
A progressive colonization procedure is described to further assess its impact on the host hepatic metabolism. Colonization is monitored non invasively by evaluating the urinary excretion of microbial co-metabolites by NMR-based metabolic profiling while hepatic metabolism is assessed by High Resolution Magic Angle Spinning (HR MAS) NMR profiling of intact biopsy.