Why Use Pre-optimized Reagents for Differentiation of Pluripotent Stem Cells into Oligodendrocytes?
Successful differentiation of pluripotent stem cells into oligodendrocytes is dependent on high quality media and differentiation supplements.
Oligodendrocytes are cells of the central nervous system which myelinate axons and support rapid nerve conduction. In CNS disorders, such as stroke, multiple sclerosis and spinal cord injury, demyelination of axons contributes to functional deficits. Studies have demonstrated that neural transplantation therapy can restore functions lost by demyelination. However, the use of neural transplantation is limited by an inadequate supply of oligodendrocyte precursors. R&D Systems offers the Mouse Oligodendrocyte Differentiation Kit to drive successful and efficient differentiation of embryonic and induced pluripotent stem cells into oligodendrocytes.
R&D Systems Mouse Oligodendrocyte Differentiation Kit:
Is pre-optimized to reduce variation.
Has defined components to decrease variation.
Includes sufficient reagents for the differentiation of 3 x 107 embryonic stem cells.
The Mouse Oligodendrocyte Differentiation Kit contains ITS and N-2 MAX Media Supplements, which are used to select and enrich neural precursor populations that are characterized by Nestin and A2B5 staining. Purified Bovine Fibronectin is included to support cell attachment and spreading. A growth factor panel, consisting of Recombinant Human Fibroblast Growth Factor Basic (FGF basic), Recombinant Human Epidermal Growth Factor (EGF), and Recombinant Human Platelet-derived Growth Factor AA (PDGF-AA) is included for effective oligodendrocyte differentiation.
The quantity of each component provided in the kit is estimated to be sufficient for the differentiation of 3 x 107 embryonic stem cells.
Recombinant Human EGF
Recombinant Human FGF basic
Recombinant Human PDGF-AA
ITS Media Supplement
N-2 MAX Media Supplement
Stability and Storage
Unopened Kit should bestored at = -20 °C in a manual defrost freezer. Do not use the kit past the expiration date.
Opened Reagents can be stored at 2 °C to 8 °C for up to 1 month or aliquotted and stored at = -20 °C in a manual defrost freezer for up to 6 months.* Avoid repeated freeze-thaw cycles.
*Provided this is within the expiration date of the kit.
Limitations of the Procedure
FOR LABORATORY RESEARCH USE ONLY. NOT FOR DIAGNOSTIC USE.
The safety and efficacy of this product in diagnostic or other clinical uses has not been established.
The quality of the embryonic stem cells and any variation in this procedure may cause variation in the efficiency of oligodendrocyte generation.
The acute and chronic effects of overexposure to reagents of this kit are unknown. Safe laboratory procedures should be followed and protective clothing should be worn when handling kit reagents.
The ITS and N-2 MAX Supplements contain human transferrin. This transferrin was tested at the donor level using an FDA licensed method and found to be non-reactive for anti-HIV-1/2 and Hepatitis B surface antigen. As no testing can offer complete assurance of freedom from infectious agents, these reagents should be handled as if capable of transmitting infection.
Mouse Oligodendrocytes Generated Using the Mouse Oligodendrocyte Differentiation Kit. D3 mouse embryonic stem cells were expanded in KO-ES Media and differentiated into oligodendrocytes using the Mouse Oligodendrocyte Differentiation Kit (Catalog # SC004). Oligodendrocytes were detected using a Mouse Anti-Human/Mouse/Rat/Chicken Oligodendrocyte Marker O4 Monoclonal Antibody (Catalog # MAB1326). The cells were stained with the NorthernLights™-557 Affinity-purified Goat Anti-Mouse Secondary Antibody (Catalog # NL019). The nuclei were counterstained with DAPI. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Neural stem cells provide an excellent model for research focused on neural development and neurological disorders. R&D Systems offers ready-to-use primary cortical stem cells isolated from E14.5 Sprague-Dawley rats. In addition, primary mouse cortical stem cells isolated from E14.5 CD-1 mice are available. Every lot of R&D Systems Cortical Stem Cells is validated for a high level of Nestin expression and the capacity for multi-lineage differentiation into astrocytes, neurons, and oligodendrocytes. Our cortical stem cells are tested to ensure highest quality and lot to lot consistency. Both rat and mouse Cortical Stem Cells can be optimally expanded as monolayers or neurospheres.
To complement the use of primary neural stem cells, we offer a range of supportive products, including culture media which is specifically optimized for use with neural stem cells. We offer kits to promote the in vitro proliferation of neural precursors, and kits to differentiate neural stem cells into dopaminergic neurons or oligodendrocytes. In addition, kits are available which contain panels of antibodies designed to monitor the differentiation and identification of neural precursors, astrocytes, neurons, and oligodendrocytes.
Mouse Oligodendrocyte Differentiation Kit Scientific Data Examples
Mouse Oligodendrocytes Generated Using the Mouse Oligodendrocyte Differentiation Kit.
D3 mouse embryonic stem cells were expanded in KO-ES Media and differentiated into oligodendrocytes using the Mouse Oligodendrocyte Differentiation Kit (Catalog # SC004). Oligodendrocytes were detected using a Mouse Anti-Human/Mouse/Rat/Chicken Oligodendrocyte Marker O4 Monoclonal Antibody (Catalog # MAB1326). The cells were stained with the NorthernLights™-557 Affinity-purified Goat Anti-Mouse Secondary Antibody (Catalog # NL019). The nuclei were counterstained with DAPI. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Preparation & Storage
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Refer to the product datasheet for complete product details.
Briefly, pluripotent stem cells are differentiated into oligodendrocytes using the following procedure:
Generate embryoid bodies from pluripotent stem cells
Select for and expand Nestin-positive cells
Induce A2B5-positive cells using supplemented media
Differentiate A2B5-positive cells into oligodendrocytes
Which Brands are Currently Available on bio‑techne.com? R&D Systems, Tocris Bioscience and ProteinSimple branded products are available to purchase through bio‑techne.com. ProteinSimple branded instruments and Simple Plex Assays are available to quote. ProteinSimple branded instrument consumables and ACD branded products will be available on bio‑techne.com in the near future. Novus Biologicals branded products are not currently available on bio‑techne.com and can be found at novusbio.com.
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