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University of California, San Francisco - UCSF

19 ARTICLES PUBLISHED IN JoVE

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Biology

Organotypic Slice Culture of E18 Rat Brains
Laura Elias 1, Arnold Kriegstein 1
1Institute for Regeneration Medicine, University of California, San Francisco - UCSF

Organotypic Slice Culture of E18 Rat Brains

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Biology

In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos
William Walantus 1, Laura Elias 1, Arnold Kriegstein 1
1Institute for Regeneration Medicine, University of California, San Francisco - UCSF

In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos

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Biology

Heterotopic Heart Transplantation in Mice
Fengchun Liu 1, Sang Mo Kang 1
1Department of Surgery, University of California, San Francisco - UCSF

The mouse heterotopic heart transplantation model has been proven by many investigators to be an important method for studying mechanisms of rejection and immune response. However, the techniques involved are still challenging. By modifying standard techniques we have had success with more than 1000 transplants.

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Biology

In Utero Intraventricular Injection and Electroporation of E15 Mouse Embryos
William Walantus 1, David Castaneda 1, Laura Elias 1, Arnold Kriegstein 1
1Institute for Regeneration Medicine, University of California, San Francisco - UCSF

In Utero Intraventricular Injection and Electroporation of E15 Mouse Embryos

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Biology

Cortical Neurogenesis: Transitioning from Advances in the Laboratory to Cell-Based Therapies
Arnold R. Kriegstein 1
1Institute for Regeneration Medicine, University of California, San Francisco - UCSF

Cortical Neurogenesis: Transitioning from Advances in the Laboratory to Cell-Based Therapies

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Biology

Murine Pancreatic Islet Isolation
Gregory L. Szot 1, Pavel Koudria 1, Jeffrey A. Bluestone 1
1Diabetes Center, University of California, San Francisco - UCSF

Murine Pancreatic Islet Isolation

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Biology

Investigating the Immunological Mechanisms Underlying Organ Transplant Rejection
Sang Mo Kang 1
1Department of Surgery, University of California, San Francisco - UCSF

Investigating the Immunological Mechanisms Underlying Organ Transplant Rejection

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Biology

Regulatory T cells: Therapeutic Potential for Treating Transplant Rejection and Type I Diabetes
Jeffry A. Bluestone 1
1Diabetes Center, University of California, San Francisco - UCSF

Regulatory T cells: Therapeutic Potential for Treating Transplant Rejection and Type I Diabetes

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Biology

Small Bowel Transplantation In Mice
Fengchun Liu 1, Sang-Mo Kang 1
1Department of Surgery, University of California, San Francisco - UCSF

The mouse small bowel transplantation model has been recognized as an important tool to study mechanismes of immune rejection and screen new immunosuppressive drugs. However, this model is limited to use because the techniques involved is an extremely technically challenge. Now we introduce the modified technique.

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Biology

Transplantation of Pancreatic Islets Into the Kidney Capsule of Diabetic Mice
Gregory L. Szot 1, Pavel Koudria 1, Jeffrey A. Bluestone 1
1Diabetes Center, University of California, San Francisco - UCSF

Our protocol was developed to cleanly and easily deliver islets or cells under the kidney capsule of mice. Cells are concentrated into pellets in the final tubing used for transplanting the cells under the kidney capsule. The ease of this technique reduces stress to the cells and the mouse.

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Biology

Orthotopic Mouse Model of Colorectal Cancer
William Tseng 1,2, Xianne Leong 2, Edgar Engleman 2
1Department of Surgery, University of California, San Francisco - UCSF, 2Department of Pathology, Stanford University School of Medicine

Two techniques can be used to establish this model: injection of a cancer cell suspension into the cecal wall or transplantation of a piece of subcutaneous tumor onto the cecum. This model is useful for studying the natural progression of colorectal cancer and testing new therapeutic agents against colorectal cancer.

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Biology

Actin Co-Sedimentation Assay; for the Analysis of Protein Binding to F-Actin
Jyoti Srivastava 1, Diane Barber 1
1Department of Cell and Tissue Biology, University of California, San Francisco - UCSF

Proteins bind to filamentous actin (F-actin) through distinct actin binding modules. In this video we demonstrate the procedure of actin co-sedimentation, which is an in vitro assay routinely used to analyze proteins or specific domains that bind F-actin.

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Biology

A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples
Eric Johansen 1, Birgit Schilling 2, Michael Lerch 1, Richard K. Niles 1, Haichuan Liu 1, Bensheng Li 2, Simon Allen 1, Steven C. Hall 1, H. Ewa Witkowska 1, Fred E. Regnier 3, Bradford W. Gibson 2, Susan J. Fisher 1, Penelope M. Drake 1
1Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco - UCSF, 2Buck Institute for Age Research, 3Department of Chemistry, Purdue University

Lectin-conjugated POROS beads were employed for HPLC. Glycopeptide standards served as positive and negative controls. MARS-14 depleted, trypsin-digested human plasma was chromatographed and flow-through (FT) and bound fractions collected for ESI-LC-MS/MS analyses. Glycopeptides were enriched in the bound fraction as compared to FT.

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Biology

Lentivirus Production
Xiaoyin Wang 1, Michael McManus 1
1Diabetes Center, University of California, San Francisco - UCSF

To make lentiviruses, DNA vectors are transfected into human 293 cells. After harvest and concentrating the supernatant, virus titer is determined by fluorescence expression with a flow cytometer.

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Neuroscience

The Subventricular Zone En-face: Wholemount Staining and Ependymal Flow
Zaman Mirzadeh 1, Fiona Doetsch 2,3, Kazunobu Sawamoto 4, Hynek Wichterle 2,5, Arturo Alvarez-Buylla 1
1Department of Neurosurgery, The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco - UCSF, 2Department of Pathology and Cell Biology, College of Physicians and Surgeons, Columbia University, 3Department of Neuroscience and Neurology, College of Physicians and Surgeons, Columbia University, 4Department of Developmental and Regenerative Biology, Nagoya City University Graduate School of Medical Sciences, 5Center for Motor Neuron Biology and Disease, College of Physicians and Surgeons, Columbia University

The lateral ventricle walls contain the largest germinal region in the adult mammalian brain. Traditionally, studies on neurogenesis in this region have relied on classical sectioning techniques for histological analysis. Here we present an alternative approach, the wholemount technique, which provides a comprehensive, en-face view of this germinal region.

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Neuroscience

Establishing Intracranial Brain Tumor Xenografts With Subsequent Analysis of Tumor Growth and Response to Therapy using Bioluminescence Imaging
Tomoko Ozawa 1, C. David James 1
1Department of Neurological Surgery, University of California, San Francisco - UCSF

Luciferase-modified human brain tumor xenografts can be established intracranially in athymic mice, with subsequent monitoring of tumor growth and response to therapy using bioluminescence imaging. In combination with survival analysis, bioluminescence monitoring is an essential research tool for pre-clinical testing of therapies being considered for treating brain tumors.

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Neuroscience

Systemic and Local Drug Delivery for Treating Diseases of the Central Nervous System in Rodent Models
Laura Serwer 1, Rintaro Hashizume 1, Tomoko Ozawa 1, C. David James 1
1Department of Neurological Surgery, University of California, San Francisco - UCSF

Thorough preclinical testing of drugs that act in the central nervous system often involves assessing and comparing drug biodistribution in association with specific routes of administration. Here, three commonly used methods of systemic delivery (intravenous, intraperitoneal, and oral) as well as a method for local delivery (convection-enhanced delivery) are demonstrated in mice.

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Developmental Biology

Viral-mediated Labeling and Transplantation of Medial Ganglionic Eminence (MGE) Cells for In Vivo Studies
Daniel Vogt 1, Pei-Rung Wu 1, Shawn F. Sorrells 2, Christine Arnold 2, Arturo Alvarez-Buylla 2, John L. R. Rubenstein 1
1Department of Psychiatry, University of California San Francisco, 2Department of Neurological Surgery, University of California San Francisco

GABAergic cortical interneuron progenitors disperse, develop and synaptically integrate into a host cortex after transplantation. These cells can be easily transduced before transplantation for in vivo studies of genetically modified GABAergic precursors. Here, we show viral labeling techniques to target specific interneuron subgroups using existing Cre lines and Cre-dependent reporters.

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Medicine

Bioluminescence Imaging of an Immunocompetent Animal Model for Glioblastoma
Aaron J. Clark 1, Shayan Fakurnejad 2, Quanhong Ma 2, Rintaro Hashizume 2,3
1Department of Neurological Surgery, University of California San Francisco, 2Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, 3Biochemistry and Molecular Genetics, Northwestern University Feinberg School of Medicine

GL261 glioma cells provide a useful immunocompetent animal model of glioblastoma. The goals of this protocol are to demonstrate proper techniques for monitoring intracranial tumor growth using in vivo bioluminescence imaging, and to verify the utility of luciferase-modified GL261 cells for studying tumor immunology and immunotherapeutic approaches for treating glioblastoma.

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