Technical Data
Media
Endothelial Progenitor Outgrowth Cells, Human, Freezing Medium
E3010-68F
Molecular Biology Grade
Storage -20°C    Shipping Dry Ice

Description:
EPOC freezing medium was optimized for cryopreservation of our human endothelium progenitor Outgrowth cells (EPOC).
This product contains fetal bovine serum. Each lot is tested for sterility.

Procedure for Freezing EPOC:
1. Freeze EPC cells when they reach 90% to 95% confluence.
2. Warm Dulbecco’s PBS (without Ca2+ and Mg2+) and 0.05% trypspin/EDTA in a 37°C water bath. Thaw EPOC freezing medium at room temperature.
3. Remove EPOC growth medium, rinse EPOC with Dulbecco’s PBS, and incubate the cells with trypsin/EDTA solution until approximately 90% of the cells begin to detach. Monitor the cells under a microscope to avoid over-trypsinization.
4. Add fetal bovine serum equal to 1/10 volume of the trypsin/EDTA solution to neutralize trypsin. Gently shake the culture vessel to mix.
5. Transfer the cell suspension into a 15ml conical tube. Take a small aliquot of the suspension to count cells. Centrifuge the cell suspension 150 x g for 5 minutes at room temperature.
6. Carefully remove the supernatant. Add a volume of EPOC freezing medium and re-suspend cells to 0.5 to 1 x 10e6 per ml. Dispense 1ml into each cryovial.
7. Place cryovials in a freezing container to slowly lower the temperature in a -80°C freezer. Transfer the vials to liquid nitrogen the next day for long term storage.

Storage and Stability:
Store at -20°C Stable for 1 year from the date of receipt under proper storage condition.



Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.