differentiation Protocols

Category: Neuroscience Protocols

Bead aggregation assay to demonstrate presynaptic differentiation induced by the NGL family of cell adhesion molecules. - [Read Bead Aggregation Assay Protocol]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Culture conditions have been established for a second blastocyst-derived cell line, trophoblast stem (TS) cells, in addition to embryonic stem (ES) cells. This protocol describes a method for culturing TS cell lines. These cells can then be used to study trophoblast differentiation and placental function. - [Read Culturing Trophoblast Stem (TS) Cell Lines Protocol]

Category: Stem Cell Protocols

Pluripotent ES cells can develop into many types of differentiated tissues if they are placed back into a differentiating environment. Often, differentiation proceeds through an intermediate stage called the embryoid body (EB). EBs can be manipulated further to generate more differentiated cell types. This protocol describes a method for differentiation of ES cells into EBs. - [Read Differentiating Embryonic Stem (ES) Cells into Embryoid Bodies Protocol]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

In vitro differentiation of ES cells occurs when the cells are allowed to aggregate in suspension culture in the absence of mouse embryonic fibroblast (MEF) feeders and leukemia inhibitory factor (LIF). Hanging drops provide a uniform aggregate size, which is then expanded by continued growth in suspension culture. The embryoid bodies are then plated and allowed to differentiate further in culture. - [Read Differentiation of Embryonic Stem (ES) Cells Using the Hanging Drop Method]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: Embryotoxic Potential Testing of Chemicals

EMBRYONIC STEM CELL TEST (EST). The embryotoxic potential of chemicals is determined by the evaluation of the inhibition of differentiation of embryonic stem cells (ES) and the inhibition of growth of ES and 3T3 cells. Scientific Information Service - [Read EMBRYONIC STEM CELL TEST (EST)]

Category: Lipid Protocols

Protocol for fatty acid oxidation assay. Includes: Differentiation of C2C12 cells; Preparation; Oleic acid oxidation. - [Read Fatty Acid Oxidation Assay Protocol]

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

This method enables the culturing of thyroid cells without loss of differentiation and medium change. It is potentially useful for the long-term study of drug effects on the thyroid gland. - [Read Human Thyroid Culture Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

The effect of a test compound, in the presence and absence of S-9 mix, on the differentiation and growth of rat limb bud and CNS cells in vitro indicates whether it is potentially a teratogen in vivo. - [Read In Vitro Micromass Teratogen Assay]

Category: RNAi Technology Protocols: RNAi Avian Embryos Protocols

Protocol describes a method for in ovo transfection of avian embryos with double-stranded RNA (dsRNA).  The dsRNA is injected into the spinal cord of the embryo.  Subsequent electroporation facilitates the cellular uptake of the dsRNA molecules.  It may be necessary to optimize the stage of the embryo and the electroporation procedure to improve the effectiveness of in ovo RNAi—cell competence changes with differentiation. - [Read Injection of dsRNA and Electroporation in Avian Embryos 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: 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: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Boris Greber. Important rules for ES cell culture, Signs of differentiation, General tips, ES medium, Thawing ES cells, Freezing, Passaging. - [Read Mouse embryonic stem (ES) cell culture - basic procedures]

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

Great Procedures. rules for ES cell culture, differentiation of ES cells, ES medium, Thawing, Passaging, Freezing. Mouse embryonic stem (ES) cell culture - basic procedures. Max Planck Institute for Molecular Genetics. Boris Greber. - [Read Mouse embryonic stem (ES) cell culture - basic procedures PDF]

Category: Neuroscience Protocols: Neuronal Cell Culture Protocols

Neuron-fibroblast coculture assay to demonstrate presynaptic differentiation.  Includes: Preparation of HEK293T cells, Coculture of HEK293T cells and neurons, Immunostaining, Vesicle turnover experiment, Image acquisition and quantitation. - [Read Neuron Fibroblast Coculture Assay to Demonstrate Presynaptic Differentiation]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Germ layers are multipotent tissues that have the ability to differentiate into various tissue types. Isolation and manipulation of germ layers is important for examining differentiation. This protocol describes a method for separating postimplantation germ layers. - [Read Separating Postimplantation Germ Layers Protocol]

Category: Staining Protocols: Simple Staining Protocols

Protocol for simple staining. Smear is stained with a solution of a single dye which stains all cells the same color. Differentiation of cell types or structures is not the objective of the simple stain. However, certain structures which are not stained by this method may be easily seen, for example, endospores and lipid inclusions. - [Read Simple Staining Protocol]

Category: Neuroscience Protocols: Neuronal Stains Protocols

Protargol-S (silver proteinate) is used with the addition of copper metal.  The copper replaces the silver in the connective tissue, allowing a greater differentiation between the nerve fibers and the connective tissue.  The silver is reduced with hydroquinone to the visible metallic form.  The sections are toned with gold chloride, the gold chloride is reduced with oxalic acid, increasing the deposit of metallic gold on the sections. - [Read Staining of Nerve Fibers Protocol]

Category: Primary Cell Isolation and Culture Protocols

Thymosin β4 (Tβ4)-induced outgrowth and differentiation of vasculogenic precursor cells. - [Read Thymosin β4 (Tβ4)-induced Outgrowth and Differentiation of Vasculogenic Precursor Cells]

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol for thymosin β4 (Tβ4)-induced outgrowth and differentiation of vasculogenic precursor cells from adult epicardium. Includes: Preparation of EPDCs; Characterisation of EPDC Phenotypes by Immunofluorescence. - [Read Thymosin β4 (Tβ4)-induced Outgrowth and Differentiation of Vasculogenic Precursor Cells Protocol]