View our list of available Coronavirus (COVID-19, SARS-CoV-2, 2019-nCOV) antibodies, recombinant proteins, ELISA Kits... Options Full Datasheet Printer Friendly Datasheet Prepare for Printing Full Datasheet Printer Friendly Datasheet Prepare for Printing Have Questions? Ask Us! Request A Quote Today! Add To Compare List SNL Feeder Cells Cell Line Catalog # MBS169018 Unit / Price 1 Vial / $895 +1 FREE 8GB USB 5x1 Vial / $3,760 +4 FREE 8GB USB Product Name Product Info Accession #s Product Desc Diseases/Tissues/Pathways Applications References Product Name SNL Feeder Cells, Cell Line Popular Item Full Product Name SNL Feeder Cells Product Synonym Names SNL Feeder Cells Research Use Only For Research Use Only. Not for use in diagnostic procedures. TOP Request for Current Manual Insert Request Current Manual MBS169018 Technical Datasheet Technical Datasheet PDF Concentration 1 mL, 3 x 10^6 cells/mL in 70% DMEM, 20% FBS, 10% DMSO (lot specific) Methods I. Establishing SNL Feeder Cell Cultures from Frozen Cells1. Place 10 ml of complete DMEM growth medium in a 50-ml conical tube. Thaw the frozen cryovial of cells by gentle agitation for 1-2 minutes in a 37 degree C water bath. Decontaminate the cryovial by wiping the surface of the vial with 70% (v/v) ethanol.2. Transfer the thawed cell suspension to the conical tube containing 10 mL of growth medium.3. Collect the cells by centrifugation at 1000 rpm for 5 minutes at room temperature. Remove the growth medium by aspiration.4. Resuspend the cells in the conical tube in 15 mL of fresh growth medium by gently pipetting up and down.5. Transfer the 15 mL cell suspension to a T-75 tissue culture flask. Place the cells in a 37 degree C incubator at 5% CO2.6. Monitor cell density daily. Cells should be passaged when the culture reaches 95% confluence.II. Freezing SNL Feeder Cells1. Trypsinize cells and resuspend cell pellet in cold Freeze Medium at twice the desired final cell concentration.2. Aliquot 1 mL of cells into sterile cryovials and place cryovials immediately into freezing container. Store overnight at -80 degree C.3. Transfer frozen vials to -135 degree C freezer or liquid nitrogen.III. Mitomycin C Treatment and Preparation of Feeder1. Culture cells to 90% confluence. Wash it once with sterile PBS.2. Add 10 ug/mL Mitomycin C (Sigma), incubate for 2 hrs.3. Wash 3 times with sterile PBS to remove Mitomycin.4. After dissociation by Trypsin, the Mitomycin-treated SNLs can be freezed and stored in liquid nitrogen, or used as feeder by plating them at 75 000 cells/cm2 in gelatin-coated tissue culture dishes for one day. Medium 1. Culture Medium: D-MEM (high glucose), 10% fetal bovine serum (FBS), 0.1 mM MEM Non-Essential Amino Acids (NEAA), 2 mM L-glutamine, 1% Pen-Strep (optional)2. Freeze Medium: 70% DMEM. 20% FBS, 10% DMSO Quality Control This cryovial contains at least 3.0 × 10^6 SNL feeder cells as determined by morphology, trypan-blue dye exclusion, and viable cell count. The SNL feeder cells are tested free of microbial contamination. Dry Ice Shipment Extra charge fee may add to your shipping cost as dry ice is required to ship this product. Preparation and Storage Liquid nitrogenNote: For best results begin culture of cells immediately upon receipt. If this is not possible, store at -80 degree C until first culture. Store subsequent cultured cells long term in liquid nitrogen.Dry Ice Shipment: Extra charge fee may add to your shipping cost as dry ice is required to ship this product.Shipping Note: Product is available for shipment in the United States, Canada and European countries. Please inquire for shipment to other countries. Other Notes Small volumes of SNL Feeder Cells cell line vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply. TOP Related Product Information for SNL Feeder Cells cell line • Immortalized cell line derived from mouse fibroblast STO cell transformed with murine LIF and neomycin resistance genes• Useful for culture of human or mouse iPS cells as well as ES cell growth• Must be mitotically inactivated prior to addition of stem cellsThe SNL 76/7 cell line, established by Dr. Allan Bradley, is clonally derived from a mouse fibroblast STO cell line transformed with neomycin resistance and murine LIF genes. SNL can be used as a feeder cell for ES cell growth, and it also has been recently used in mouse or human iPS culture.Background: Embryonic stem (ES) cells have been derived from the inner cellmasses (ICM) of blastocysts in many species. They are capable of unlimited, undifferentiated proliferation on feeder cell layers and remain karyotypically normal and phenotypically stable. In addition, ES cells have the ability to differentiate into a wide variety of cell types in vitro and in vivo. In mES cell culture, the feeder layer can be replaced by the addition of LIF in the growth medium. However, LIF does not have the same effect on hES cell culture as mES. Therefore, both the derivation and maintenance of hES cells require the use of feeder cells.SNL 76/7, established by Dr. Allan Bradley (1), is clonally derived from a mouse fibroblast STO cell line transformed with neomycin resistance and murine LIF genes. SNL can be used as a feeder cell for ES cell growth, and it also has been recently used in mouse or human iPS culture (2, 3, 4). Product Categories/Family for SNL Feeder Cells cell line Stem Cell Research; Feeder Cells; SNL 76/7 Passage-Independent Feeder Cells Application Notes for SNL Feeder Cells cell line SNL feeder cells are used for the maintenance of ES or iPS cells in the undifferentiated state. The cells must be mitotically inactivated prior to the addition of ES or iPS cells, such as treatment with mitomycin C (2-4 hr, 10 ug/mL). TOP Product References and Citations for SNL Feeder Cells cell line 1. McMahon, A.P. and Bradley, A. (1990) Cell 62:1073-1085.2. Okita, K; Ichisaka, T; Yamanaka, S. (2007) Nature 448:313-317.3. Takahashi K, Okita K, Nakagawa M, Yamanaka S. (2007) Nat Protoc. 2:3081-9.4. Takahashi K, Narita M, Yokura M, Ichisaka T, Yamanaka S. (2009) PLoS One 4(12):e8067.1. Yi, L. et al. (2012). Multiple Roles of p53-Related Pathways in Somatic Cell Reprogramming and Stem Cell Differentiation. Cancer Res. 72: 5635-5645. TOP Precautions All of MyBioSource's Products are for scientific laboratory research purposes and are not for diagnostic, therapeutics, prophylactic or in vivo use. Through your purchase, you expressly represent and warrant to MyBioSource that you will properly test and use any Products purchased from MyBioSource in accordance with industry standards. MyBioSource and its authorized distributors reserve the right to refuse to process any order where we reasonably believe that the intended use will fall outside of our acceptable guidelines. Disclaimer While every efforts were made to ensure the accuracy of the information provided in this datasheet, MyBioSource will not be liable for any omissions or errors contained herein. MyBioSource reserves the right to make changes to this datasheet at any time without prior notice.It is the responsibility of the customer to report product performance issues to MyBioSource within 30 days of receipt of the product. Please visit our Terms & Conditions page for more information. TOP