mouse Protocols

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Human embryonic stem cells (hESCs) are pluripotent cells capable of differentiation to representatives of all three germ layers. Includes: Isolation of Primary Mouse Embryo Fibroblasts; Thawing and preparing p1 MEF feeder plates; Preparation of MEF- Conditioned Medium (MEF-CM; Microdissection Passaging of hESCs; Bulk passaging of hESC; Cryopreservation of hESCs; Thawing of hESCs; Karyotyping. - [Read Human Embryonic Stem Cell Protocols]

Category: Cytoskeleton Protocols: Actin Myosin Protocols

The same GFP-tagged actin construct used in cell transfection experiments has been used to produce transgenic mice. Transgenic animals allow the imaging of brain tissue in the intact animal, as acutely cut slices or as organotypic slice cultures. They also serve as a source of cells for imaging neurons at high resolution in dispersed low-density cell culture. In contrast to cells transfected in culture, where the level of actin-GFP expression in neurons varies considerably, transgenic mice... - [Read Imaging Actin in Tissue Slices from Transgenic Mouse Brain Protocol]

Category: Immunology: Immunofluorescence Protocols: Immunofluorescence Staining Protocols

Protocol for immunofluorescent staining of cell suspensions of mouse lymphoid tissue. - [Read Immunofluorescent Staining of Cell Suspensions of Mouse Lymphoid Tissue Protocol]

Category: Mouse Protocols

Protocol for the immunohistology of mouse spleens. - [Read Immunohistology of Mouse Spleens Protocol]

Category: Stem Cell Protocols: Stem Cell Injection Protocols

This protocol describes a method for injecting mouse blastocysts with embryonic stem (ES) cells to produce chimeras. - [Read Injecting Blastocysts Protocol]

Category: Mouse Protocols: Mouse Surgical Procedures Protocols

Protocol describes intraperitoneal (IP) injection, which is the typical means of introducing most compounds, such as hormones and anesthetics, into the mouse. - [Read Intraperitoneal (IP) Injection Protocol]

Category: Mouse Protocols: Mouse Embryology Protocols

Isolation of extraembryonic tissues allows one to study normal mouse development as well as the molecular basis of defects which cause fetal death. This protocol describes a method for isolating extraembryonic membranes from pregnant mice. - [Read Isolating Mice Extraembryonic Membranes Protocol]

Category: Immunology: Mononuclear Cell Isolation Protocols

Mononuclear phagocyte progenitor cells derived from femoral and tibial bone marrow are propagated in the presence of M-CSF. This macrophage growth factor is secreted by L929 cells and is used in the form of L929 cell conditioned medium. The progenitor cells proliferate and differentiate through monoblast, promonocyte and monocyte stages before maturing to macrophages. At this time the cells become firmly adherent to the culture vessel. - [Read Isolation and Culture of Mouse Bone Marrow-Derived Macrophages Protocol]

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the isolation of fibroblasts from mouse embryos. Mouse embryonic fibroblast (MEF) cells are used as a feeder layer for the culture of mouse embryonic stem (ES) cells to help maintain them as pluripotent stem cells. The inhibition of ES-cell differentiation provided by the MEF feeders appears to be due to their production of leukemia inhibitory factor (LIF). - [Read Isolation and Freezing of Primary Mouse Embryonic Fibroblasts (MEF) For Feeder Plates]

Category: Mouse Protocols: Mouse Cell Culture Protocols

The procedures involve the isolation and growth of primary cell cultures from rodent and human tissue as well as the use of viral vectors for the introduction and expression of mammalian genes in cells in culture and in live rodents. - [Read Isolation and Growth of Primary Cell Cultures from Mouse Protocol]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: MEF Protocols Mouse Embryonic Fibroblasts

Embryonic stem cells are usually grown on a layer of mitotically inactivated primary mouse embryonic fibroblasts to promote growth and prevent differentiation. Boris Greber - [Read Isolation and handling of primary mouse embryonic fibroblasts (MEFs)]

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

This procedure describes the isolation and culture of adult mouse cardiac myocytes from one heart. The isolation routinely yields approximately 1 million rodshaped myocytes per heart. - [Read Isolation of Adult Mouse Cardiac Myocytes from One Heart Protocol]

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol describes the isolation and culture of adult mouse cardiac myocytes from one heart. The isolation routinely yields approximately 1 million rodshaped myocytes per heart. - [Read Isolation of Adult Mouse Cardiac Myocytes from One Heart Protocol]

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

This procedure describes the isolation and culture of adult mouse cardiac myocytes from two or more hearts. Includes modifications for the digestion of two or more hearts in the same procedure and subsequent pooling of myocytes derived from the multiple hearts. The isolation procedure is performed by one or more technicians and routinely yields approximately 1 million rod-shaped myocytes per heart. - [Read Isolation of Adult Mouse Cardiac Myocytes from Two or More Hearts Protocol]

Category: Immunology: Mononuclear Cell Isolation Protocols

Presents two methods for preparing dendritic cells (DCs), a highly specialized type of antigen-presenting cell (APC). The first method involves the isolation of DCs from mouse spleen, resulting in a cell population that is highly enriched in accessory cell and APC function. A support protocol for collagenase digestion of splenocyte suspensions is described to increase the yield of dendritic cells. The second method involves generating large numbers of DCs from mouse bone marrow progenitor cells. - [Read Isolation of Dendritic Cells Protocol]

Category: Transgenic Mouse Protocols: Mouse Tail DNA Extraction Genotyping

Isolation of DNA from Mouse Tail Biopsies. University of Michigan Transgenic Animal Model Core, Transgenic Animal Web. - [Read Isolation of DNA from Mouse Tail Biopsies]

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

This protocol describes the isolation of fibroblasts from mouse, rat, and hamster embryos. - [Read Isolation of Fibroblasts from Mammalian Embryos Protocol]

Category: Mouse Protocols: Mouse Embryology Protocols

Isolation of mouse embryos video clip. - [Read Isolation of Mouse Embryos Video Clip]

Category: Mouse Protocols

Provides two protocols that may be used to isolate IHL. One can be used to isolate IHL from multiple livers in parallel, whereas the more elaborate alternate protocol yields more cells per liver but is more appropriately used to recover the IHL from a single liver. - [Read Isolation of Mouse Intrahepatic Lymphocytes Protocol]

Category: Immunology: Mononuclear Cell Isolation Protocols

Covers techniques for preparing lymphoid cell suspensions and removing red blood cells and dead cells. - [Read Isolation of Mouse Mononuclear Cells Protocol]

Category: Immunology: T Cell Protocols

Peyer’s Patch, and Lamina Propria Cells lymphocyte populations should be analyzed when studying the immunological status of the intestine, for example in oral immunization or in intestinal disease (including infectious disease and tumors). This protocol details techniques for isolation of IEL, PP cells, and LP cells from the small intestine of the mouse. - [Read Isolation of Mouse Small Intestinal Intraepithelial Lymphocytes Protocol]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Isolation of Primary Fibroblasts from Mouse Embryos. Bradley's Lab, Texas. - [Read Isolation of Primary Fibroblasts from Mouse Embryos]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: MEF Protocols Mouse Embryonic Fibroblasts

Isolation of Primary Mouse Embryonic Fibroblasts (PMEFs). ESGRO. - [Read Isolation of Primary Mouse Embryonic Fibroblasts (PMEFs)]

Category: Immunology: B Cell Protocols

Describes the isolation of resting B lymphocytes (B cells) from mouse spleens by using negative selection with anti-CD43 and anti-Mac-1/CD11b monoclonal antibodies coupled to magnetic microbeads. This strategy depletes non-B cells from a mixed population of splenocytes and relies on the fact that most mature leukocytes, with the exception of resting splenic B cells, express CD43. - [Read Isolation of Resting B Lymphocytes from One or More Groups of Four Mouse Spleens Protocol]

Category: Immunology: Mononuclear Cell Isolation Protocols: B Lymphocyte Isolation Protocols

Protocol describes the isolation of resting B lymphocytes (B cells) from mouse spleens by using negative selection with anti-CD43 and anti-Mac-1/CD11b monoclonal antibodies coupled to magnetic microbeads. This strategy depletes non-B cells from a mixed population of splenocytes and relies on the fact that most mature leukocytes, with the exception of resting splenic B cells, express CD43. - [Read Isolation of Resting B Lymphocytes from Sixteen Mouse Spleens Protocol]