SCM014 Neural Stem Cell Freezing Medium (1X)

SCM014
50 mL  
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      Overview

      Replacement Information

      Key Specifications Table

      Key Applications
      Cult
      Description
      Catalogue NumberSCM014
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionNeural Stem Cell Freezing Medium (1X)
      References
      Product Information
      PresentationSerum-free formulation. Contains 10% DMSO.
      Applications
      ApplicationThe Neural Stem Cell Freezing Medium (1X) has been optimized & validated for Stem cell culture. It is available in a 50ml format.
      Key Applications
      • Stem Cell Culture
      Application Notes1. Thaw cell culture freezing medium completely and mix well by gently swirling bottle. Keep freezing medium on ice during use.

      2. Cells to be frozen should be in late log phase growth.

      3. Monolayers will need to be dissociated. After dissociation, cells are resuspended in Neural Stem Cell Basal Medium (Cat. No. SCM003) and counted to determine viability and number.

      4. Centrifuge cells at 300 x g for 3 min. Remove the medium above the pellet.

      5. Resuspend the cells in Neural Stem Cell Freezing Medium at a concentration of ~4 x 106 cells/mL. Freeze 1 mL of cells/vial. After the cells have been resuspended and aliquoted into appropriate cryogenic storage vials, they can be placed in a freezing container and the normal freeze down procedure should be started within five minutes.

      6. Cells must be stored at or below -80°C. For long term storage the cells should be stored in ultra-low temperature freezer (-150°C), or in liquid nitrogen (-196°C).

      7. Thawing of cryopreserved cells should be as follows:

      a. Do not thaw the cells until the recommended medium and appropriately coated poly-L-ornithine and laminin plasticware and/or glassware are on hand.
      b. Thaw cells quickly in a 37°C water bath. Important: Do not vortex the cells.
      c. Sterilize vial by rinsing with 70% ethanol.
      d. In a laminar flow hood, use a 1 or 2 mL pipette to transfer the cells to a sterile 15 mL conical tube. Be careful to not introduce any bubbles during the transfer process.
      e. Using a 10 mL pipette, slowly add dropwise 9 mL of Neural Stem Cell Basal Medium (pre-warmed at 37°C) to the 15 mL conical tube. IMPORTANT: Do not add the whole volume of medium at once to the cells. This may result in decreased cell viability due to osmotic shock.
      f. Gently mix the cell suspension by slow pipeting up and down twice. Be careful to not introduce any bubbles. IMPORTANT: Do not vortex the cells.
      g. Centrifuge the tube at 300 x g for 2-3 minutes to pellet the cells.
      h. Decant as much of the supernatant as possible
      i. Resuspend the cells in a total volume of 10 mL of Neural Stem Cell Basal Medium (pre-warmed at 37°C) containing freshly added growth factors.

      Note: Growth factors should always be added fresh to the Neural Stem Cell Basal Medium. For rat neural stem cells: add 2-4 μL of FGF-2 (100 μg/mL) stock to 10 mL of Neural Stem Cell Basal Medium. For mouse neural stem cells, add 2 μL of FGF (100 μg/mL stock), EGF (100 μg/mL stock) and heparin (10 mg/mL stock) to 10 mL of Neural Stem Cell Basal Medium.

      j. Plate the cell mixture onto a poly-L-ornithine and laminin-coated 10-cm tissue culture plate.
      k. Incubate the cells at 37°C in a 5% CO2 humidified incubator.
      l. The next day, exchange the medium with fresh Neural Stem Cell Basal Medium (pre-warmed to 37°C) containing freshly added growth factors (refer to step 7i for growth factor concentrations). Exchange with fresh medium containing growth factors every other day thereafter.
      m. When the cells are approximately 80% confluent, they can be dissociated with Accutase™ (Cat. No. SCR005) and passaged or alternatively frozen for later use.
      Biological Information
      Media FormLiquid
      Stem Cell Type
      • Neural Stem Cells
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStore at -20°C.
      Packaging Information
      Material Size50 mL
      Transport Information
      Supplemental Information
      Specifications