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1. to determine the viable cell density 2 Centrifuge the cells for 3 5 minutes at 250 x g and 20 C Remove and discard the IMPIOOO3A2 NCPF 10001 Page 1 of 3 oe mene cp cyagen Supernatant using a pipette 3 Resuspend the cell pellet in the OriCelITM NCR Protein Free Cryopreservation Medium at a cell density of 10 10 cells mL 4 Dispense aliquots of the cell suspension into cryogenic storage vials that are properly labeled 5 Place the vials directly in a 80 C freezer After 24 hours transfer the frozen vials to liquid nitrogen for long term preservation Thawing 1 Remove the cryovial of frozen cells from storage and quickly thaw the vial in a 37 C water bath 2 Gently dilute and resuspend the cells in 8 10mL of cell culture medium 3 Centrifuge the cells for 3 5 minutes at 250 x g and 20 C 4 Carefully aspirate off as much of the supernatant as possible and add 2 3mL of cell culture medium to resuspend the cells 5 Transfer the cells into an appropriate growth vessel with the appropriate amount of growth medium and incubate inside a 37 C incubator with a 5 CO humidified atmosphere STABILITY AND STORAGE Store at 2 8 C This product is stable at 2 8 C for up to 3 years and should be discarded beyond the labeled expiration date For optimal performance repeated warming and freeze thawing should be avoided QUALITY CONTROL OriCell NCR noncontrolled rate Protein Free Cryopreservation Medium h2. as been tested for performance on mesenchymal stem cells and Vero cells The standard evaluation includes e Sterility test bacteria fungi and mycoplasma e pH test e Osmolality IMP10003A2 NCPF 10001 Page 3 of 4 SO gus cy cyagen e Endotoxin Cyagen Biosciences reserves all rights on the technical documents of its OriCell cell culture products No part of this document may be reproduced or adapted for other purposes without written permission from Cyagen Biosciences KAAERHETIV F DS IP VIVNAAATAAIVADNBA T 564 0063 ARAKEA LRT 118435 KYUHO LIRE Jvaf EER HAS TEL 06 6990 8051 FAX 06 6325 6058 FI AILYAR k TEL 072 636 8160 FAX 072 634 7222 http www dspbio co jp dspb ls bio ds pharma co jp IMP10003A2 NCPF 10001 Page 4 of 4
3. fA DS PHARMA IY BIOMEDICAL eee J ya g ji We help You discover life OriCell NCR Protein Free Cryopreservation Medium Cat No NCPF 10001 c DS PHARMA 7 BIOMEDICAL Gyagen PRODUCT DESCRIPTION OriCell NCR noncontrolled rate Protein Free Cryopreservation Medium is a serum free and ready to use freezing medium Its chemically defined and protein free formulation has been optimized to stem cells and primary cells thus greatly enhancing the viability and integrity of these cells by protecting them from damage during the one step freeze thaw procedure Unlike other conventional freezing media which require a slow programmed freeze our product allows the cells to be resuspended and put directly at 80 C This product is intended for laboratory research use only It is not intended for diagnostic therapeutic clinical household or any other applications FEATURES e Stable and ready to use e Chemically defined and protein free e Promotes high cell viability gt 90 for most mammalian cells post thaw e Maintain stem cell pluripotency e Does not affect the proliferation and differentiation properties of cells e Cells can be directly frozen at 80 C thus eliminating the need for a programmed freeze PACKAGING Available in two sizes 50 mL Bottle 20 mL Bottle INSTRUCTIONS Cryopreservation 1 Collect cells that are in the logarithmic growth phase Perform a cell count
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