1Institute for Physiological Chemistry, Department of Biochemical Endocrinology, University of Duisburg-Essen, 2Institute for Anatomy, Department of Neuroanatomy, University of Duisburg-Essen, 3Morphoplant GmbH, 4ARCONS Institute for Applied Research and Didactics
The chick chorioallantoic membrane (CAM) is a unique, naturally immunodeficient supportive culture environment to study angiogenesis and tumorigenesis. This video article demonstrates the different steps in chick ex ovo culture, application of potentially angiogenic substances and successful inoculation of tumor cells and tissues on the surface of the CAM.
Published November 30, 2009. Keywords: Developmental Biology, chick, ex ovo culture, in ovo culture, chorioallantoic membrane, CAM, angiogenesis, tumor, limb buds; grafting
1Neuroscience Division of the Biomedical & Health Sciences Institute, University of Georgia, 2Department of Cellular Biology, University of Georgia
Serum utilized in embryo cultures contains unknown components that can affect the outcome of experiments especially in studies involving signaling interactions. Here we utilized a serum-free oxygenated culture system and show that mid-gestation mouse embryos cultured for 16-40 hr exhibit morphological development comparable to embryos developing in utero.
Published March 1, 2014. Keywords: Developmental Biology, mouse embryo, mid-gestation, serum-free, defined media, roller culture, organogenesis, development
1Department of Evolutionary Functional Genomics, Evolutionary Biology Center, Uppsala University, 2Department of Plant Biology and Forest Genetics, Uppsala BioCenter, Swedish University of Agricultural Sciences
We describe a modified DIG in situ hybridization protocol, which is fast and applicable on a wide range of plant species including Norway spruce. With just a few adjustments, including altered RNase treatment and proteinase K concentration, the protocol may be used in studies of different tissues and species.
Published April 17, 2009. Keywords: Plant Biology, RNA, expression analysis, Norway spruce, Arabidopsis, rapeseed, conifers
1Program in Epithelial Biology, Stanford University School of Medicine
Tissue-specific analysis of a hair follicle regeneration model using lentivirus to mediate gain- or loss-of-function.
Published February 28, 2013. Keywords: Genetics, Tissue Engineering, Medicine, Biomedical Engineering, Cellular Biology, Surgery, Epithelial Biology, regeneration, chamber, hair, follicle, dermis, dermal cells, keratinocyte, graft, epithelial, cell culture, lentivirus, knockdown, shRNA-mediated knockdown, overexpression, mice, transgenic mice, animal model
1Department of Biochemistry and Cell Biology, Rice University
An approach for analyzing migration and eventual fate of avian neural crest cells in quail-chick chimeric embryos is described. This method is a simple and straightforward technique for tracing neural crest cells during migration and differentiation that are otherwise difficult to distinguish within an unmanipulated chick embryo.
Published February 7, 2012. Keywords: Neuroscience, Neural crest, chick, quail, chimera, fate map, cell migration, cell differentiation
JoVE Clinical and Translational Medicine
1Department of Pathology, University of Alabama at Birmingham - UAB, 2Department of Radiology, University of Alabama at Birmingham - UAB, 3Department of Cell Biology and Neurobiology, University of Alabama at Birmingham - UAB
A method for reliably grafting luciferase-tagged human malignant peripheral nerve sheath tumor cells into the sciatic nerve of immunodeficient mice is described. The use of bioluminescence imaging to demonstrate proper establishment of tumor grafts and criteria for random segregation of animals into study groups are also discussed.
Published March 7, 2011. Keywords: Medicine, Orthotopic grafting, Schwann cell, sciatic nerve, MPNST, neurofibrosarcoma, neurofibromatosis, experimental therapeutics
1Research Department of Cell and Developmental Biology, University College London
Slice cultures facilitate the manipulation of embryo development by gene and pharmacological perturbations. However, culture conditions must ensure that normal development can proceed within the reduced environment of the slice. We illustrate a protocol that facilitates normal spinal cord development to proceed for at least 24 hr.
Published March 25, 2013. Keywords: Developmental Biology, Neurobiology, Neuroscience, Medicine, Cellular Biology, Molecular Biology, Anatomy, Physiology, Biomedical Engineering, Genetics, Surgery, Cells, Animal Structures, Embryonic Structures, Nervous System, spinal cord, embryo, development, Slice-Culture, motor neuron, neurons, immunostaining, chick, imaging, animal model
JoVE Clinical and Translational Medicine
1Department of Surgery, University Hospital Zürich, 2Zürich Centre for Integrative Human Physiology, University of Zürich, 3Institute of Veterinary Physiology, Vetsuisse Faculty, University of Zürich, 4Imperial Weight Centre, Department of Investigative Medicine, Imperial College London
Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool.
Published June 11, 2012. Keywords: Medicine, Physiology, Roux-en-Y Gastric bypass, rat model, gastric pouch size, gut hormones
1Laboratory of Stem Cell and Neuro-Vascular Biology, Genetics and Developmental Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health
We introduce a whole-mount immunohistochemistry and laser scanning confocal microscopy with multiple labelling for analyzing intricate vascular network formation in mouse embryonic limb skin.
Published May 20, 2011. Keywords: Developmental Biology, Confocal microscopy, whole-mount immunohistochemistry, mouse embryo, blood vessel, lymphatic vessel, vascular patterning, arterial differentiation
1Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, Keck School of Medicine, University of Southern California
We present a simple protocol to visualize regions of programmed cell death (PCD) in mouse embryos and differentiating embryonic stem (ES) cell cultures using a highly soluble dye called LysoTracker.
Published October 11, 2012. Keywords: Developmental Biology, Molecular Biology, Stem Cell Biology, Cellular Biology, mouse embryo, embryonic stem cells, lysosome, programmed cell death, imaging, sonic hedgehog