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        Cell Culture Media and Solutions

        互联网

        1803

         

         

        165.0 ml fetal bovine serum, heat inactivated

        12.0 ml 200mM (100X) L-glutamine

        12.0 ml antimycotic/antibiotic (100X), liquid, Gibco, Cat. No. 600-5240AG

        Filter sterilize through a 0.22 � cellulose acetate filter and store up to 2 weeks at 4 degrees C.

         

        1X Cyclosporin media: (100 ml)

         

        1.0 ml 100X Cyclosporin A

         

        2X Cyclosporin A media: (1 �/ml)

         

         

        100X Cyclosporin A: (100ml)

         

         

        Freezing media: (1 liter)

         

        Store the tubes at -80 degrees C for up to one year.

        700 ml RPMI-1640 with 2mM L-Glutamine

        200 ml fetal bovine serum (FBS)

        100 ml dimethyl sulfoxide (DMSO, Sigma)

        --------

        1000 ml total volume

        Note: Filter sterilize media and FBS with a 0.22 � cellulose acetate filter. Do NOT filter DMSO, it will dissolve the cellulose acetate membrane.

         

        Growth media: (600 ml)

         

        90.0 ml FBS

        6.0 ml 200 mM (100X) L-glutamine

        0.6 ml 50 mg/ml gentamicin reagent

        Note: Filter sterilize through a 0.22 � filter and store up to 2 weeks at 4 degrees C.

         

        Growth media: (1 liter)

         

        165.0 ml fetal bovine serum, heat inactivated at 50-60 degrees C for one half hour

        12.0 ml 200mM (100 X) L-glutamine

        1.2 ml 50 mg/ml gentamicin reagent

        Filter sterilize through a 0.22 � cellulose acetate filter and store up to 2 weeks at 4 degrees C.

         

        Wash media: (1 liter)

         

        10.0 ml 2.5 M (100X) Hepes buffer

        1.2 ml 50 mg/ml gentamicin reagent

        Filter sterilize through a 0.22 � cellulose acetate filter and store up to 2 weeks at 4 degrees C.

         

         

          Antimycotic/antibiotic media:
          • To 1 liter of sterile RPMI 1640 with 2mM L-glutamine, add:
            To 100 ml of growth media add:
            To 100 ml of growth media add 2 ml of 100X Cyclosporin A.
            Dissolve 1 mg CSA in 0.1 ml ethanol in a sterile 15 ml centrifuge tube with a small magnetic stirrer. Add 0.02 ml (= 20 � ) of Tween 80 and mix well. While continually stirring, add 1 ml RPMI drop by drop. Bring to a final volume of 100 ml with RPMI. Filter sterilize with a 0.22 � filter. Store at 4 degrees C for up to 4 months.
            Prepare a 1 liter volume and divide into 25-50 ml centrifuge tubes containing 40 ml each.
            To 500 ml of sterile RPMI 1640 with 2 mM L-glutamine, add:
            To 1 liter of strerile RPMI 1640 with 2mM L-glutamine, add:
            To 1 liter of sterile RPMI 1640 with 2mM L-glutamine, add:

         

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