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Preparation of NeoR Murine Embryonic Fibroblasts Protocol - http://ourworld.compuserve.com/homepages/TheBroons/tnp7.htm

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

Protocol for the preparation of NeoR murine embryonic fibroblasts. - [Read Preparation of NeoR Murine Embryonic Fibroblasts Protocol]

Preparation of NeoR Murine Embryonic Fibroblasts Protocol - http://ourworld.compuserve.com/homepages/TheBroons/tnp7.htm

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol for the preparation of NeoR murine embryonic fibroblasts. - [Read Preparation of NeoR Murine Embryonic Fibroblasts Protocol]

Preparing Embryonic Stem (ES) Cells for Aggregation Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4419

Category: Stem Cell Protocols

Ths protocol describes the preparation of ES cells, which can then be aggregated with diploid or tetraploid embryos to produce aggregation chimeras. - [Read Preparing Embryonic Stem (ES) Cells for Aggregation Protocol]

Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4400

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the preparation of feeder cells from MEF cells or from the STO mouse fibroblast cell line. The cells are rendered mitotically inactive by treatment with {gamma}-irradiation. The feeder layers can then be used to maintain embryonic stem (ES) cells in the undifferentiated state. - [Read Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells Protocol]

Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomyc - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4399

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the preparation of feeder cell layers from MEF cells or from the STO mouse fibroblast cell line. The cells are rendered mitotically inactive by treatment with mitomycin C. The feeder layers can then be used to maintain embryonic stem (ES) cells in the undifferentiated state. - [Read Preparing Feeder Cell Layers from STO or Mouse Embryo Fibroblast (MEF) Cells: Treatment with Mitomyc]

Preparing Mouse Embryo Fibroblasts Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4398

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the preparation of mouse embryo fibroblasts (MEFs), which can then be used as feeder cells to maintain embryonic stem (ES) cells in the undifferentiated state. - [Read Preparing Mouse Embryo Fibroblasts Protocol]

Preparing Siliconized Pipettes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4355

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol describes a method for preparing siliconized pipettes. Such pipettes minimize the loss of embryos or embryonic tissues during transfer. - [Read Preparing Siliconized Pipettes Protocol]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

This protocol describes the culture of embryonic stem (ES) cells using mitotically inactivating primary mouse embryonic fibroblast (MEF) cells as a feeder layer (preparation described in Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates). The ES culture medium is supplemented with recombinant leukemia inhibitory factor (LIF) to help maintain the cells as pluripotent stem cells. This protocol has been optimized for the ES-D3 cell line. - [Read Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol]

Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4484

Category: Stem Cell Protocols: Mouse Embryonic Fibroblasts Protocols

Protocol for Nuclear Staining of Plants for Confocal Microscopy - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4685

Category: Microscopy Protocols: Confocal Microscopy Protocols

Protocol describes a useful way to observe the development of embryos, as well as meristems & young primordia developing at the shoot apex by confocal microscopy after staining the nuclei with propidium iodide. The number of cells can be exactly quantified in a meristem or in young primordia. Because embryonic & meristematic cells are largely filled out by their nuclei, it is easier to image only the nuclei. This method allows analysis of whole-mount material, which is more easily reconstructed. - [Read Protocol for Nuclear Staining of Plants for Confocal Microscopy]

Protocol Live Imaging of Caenorhabditis Elegans - http://www.cshprotocols.org/cgi/content/extract/2006/29/pdb.ip19

Category: Microscopy Protocols: In Vivo Imaging Protocols

To image early cleavages and chromatin dynamics, it is convenient to use histone H2B fused to GFP or lamin::GFP. Time-lapse movies can be obtained using conventional confocal microscope systems and their included software. Early embryos dissected from transgenic hermaphrodites are placed with egg salts on agar pads. Chromatin dynamics can be followed easily, and wild-type embryonic cells can be compared with mutants or RNAi-treated embryos. - [Read Protocol Live Imaging of Caenorhabditis Elegans]

Protocols for the Maintenance of Human Embryonic Stem Cells in Feeder Free Conditions - http://www.geron.com/PDF/scprotocols.pdf

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

Includes protocols: Mouse Embryonic Fibroblasts (MEF) Preparation; Harvesting MEFs; Cryopreservation of MEFs; Thawing and maintaining MEFs; Irradiating & Plating MEFs; Culture of Human ES cells with Matrigel® and Conditioned Medium; Preparation of Conditioned Medium (CM); Preparation of Matrigel® -coated plates; Passage of human ES cells on Matrigel®; Daily maintenance of feeder-free culture; Freezing Human ES Cells; Thawing Human ES cells; Formation of Embryoid Bodies; - [Read Protocols for the Maintenance of Human Embryonic Stem Cells in Feeder Free Conditions]

Simplifying hESC culture - http://www.bloodjournal.org/cgi/content/full/105/12/4550

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Growth Media and Requirements

Human embryonic stem cells are a valuable resource for research and cell replacement therapy but are notoriously cumbersome to culture. Bhatia and colleagues show that an increased dose of basic fibroblast growth factor eliminates the need for feeder laye - [Read Simplifying hESC culture]

Subcutaneous Injection of Embryonic Stem (ES) Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4383

Category: Stem Cell Protocols: Stem Cell Injection Protocols

This protocol describes a method for subcutaneous injection of ES cells into mice. - [Read Subcutaneous Injection of Embryonic Stem (ES) Cells Protocol]

Techniques for Primary Culture of C. elegans Embryonic Neurons - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=14

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: C. elegans Neuronal Cell Culture Protocols

Development of techniques for primary culture of C. elegans embryonic neurons. - [Read Techniques for Primary Culture of C. elegans Embryonic Neurons]

Techniques for Primary Culture of C. elegans Embryonic Neurons II - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=15

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: C. elegans Neuronal Cell Culture Protocols

Development of techniques for primary culture of C.elegans embryonic neurons. - [Read Techniques for Primary Culture of C. elegans Embryonic Neurons II]

Techniques for Primary Culture of C. elegans Embryonic Neurons III - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=16

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols: C. elegans Neuronal Cell Culture Protocols

Development of techniques for primary culture of C. elegans embryonic neurons - [Read Techniques for Primary Culture of C. elegans Embryonic Neurons III]

Tissue subcellular fractionation and protein extraction for use in mass-spectrometry-based proteomic - http://www.nature.com/nprot/journal/v1/n4/abs/nprot.2006.273.html

Category: Protein Protocols: Protein Extraction From Tissue

Tissue subcellular fractionation and protein extraction for use in mass-spectrometry-based proteomics. Although many methods exist for fractionating proteins, the method described here can capture the majority of subcellular fractions simultaneously at reasonable purity. The scalability of this method makes it amenable to small samples, such as embryonic tissues, in addition to larger tissues. The protocol described is for the general fractionation and extraction of proteins from organs / tissue - [Read Tissue subcellular fractionation and protein extraction for use in mass-spectrometry-based proteomic]

Transplantation of Tissues Under the Kidney Capsule Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4382

Category: Mouse Protocols: Mouse Surgical Procedures Protocols

Protocol describes a method for transplanting tissue under the kidney capsule. The in vivo developmental potential of transplanted adult or embryonic tissues can then be determined. - [Read Transplantation of Tissues Under the Kidney Capsule Protocol]

Treat MOUSE EMBRYONIC FIBROBLASTS (MEFs) with Mitomycin C - http://www.uphs.upenn.edu/abramson/simonPlasmids.html

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

Treat MOUSE EMBRYONIC FIBROBLASTS (MEFs) with Mitomycin C - [Read Treat MOUSE EMBRYONIC FIBROBLASTS (MEFs) with Mitomycin C]

Articles (1-2 of 2)

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

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