-------引自Clonetics® Endothelial Cell Systems (Instructions for Use)
Unpacking and Storage Instructions
4. BulletKit® Instructions: Upon arrival, store Basal Medium at 4-8°C and SingleQuots® at -20°C in a freezer that is not self-defrosting. If thawed upon arrival, growth factors can be stored at 4°C and added to basal medium within 72 hours of receipt. After SingleQuots® are added to basal medium, use within one month. Do not re-freeze.
5. ReagentPack™ Subculture Reagents are sterile-filtered and then stored at -20°C until shipment. Subculture reagents may thaw during transport. They may be refrozen once. If you plan to use within 3 days, store at 4°C. Trypsin/EDTA Solution has a limited shelf life or activation at 4°C. If, upon arrival, Trypsin/EDTA is thawed, immediately aliquot and refreeze at –20°C. We recommend that the HEPES-BSS and the Trypsin Neutralizing Solution be stored at 4°C for no more than one month. Note: To keep Trypsin/EDTA fresh and active after thawing, you may aliquot it into sterile centrifuge tubes and re-freeze at -20°C.
Preparation of Medium（Perform the following steps in a sterile field. ）
For EGM® Complete, do the following:
1. Decontaminate the vial of BBE and medium bottle with ethanol or isopropanol.
2. Add the entire contents of the vial (approximately 2 ml) to the media with a pipette. Rinse the BBE vial with medium and pipette the contents back into the 500 ml bottle.
3. Replace the cap and swirl the medium gently a few times to mix.
4. Record the date the BBE was added on the medium label.
For a BulletKit®, perform the following steps:
1. Decontaminate the external surfaces of all supplement vials and the medium bottle with ethanol or isopropanol.
2. Aseptically open each supplement vial and add the entire amount to the basal medium with a pipette.
3. Rinse each cryovial with the medium. It may not be possible to recover the entire volume listed for each cryovial. Small losses, even up to 10%, should not affect the cell growth characteristics of the supplemented medium.
4. Transfer the label provided with each kit to the basal medium bottle being supplemented. Use it to record the date and amount of each supplement added. We recommend that you place the completed label over the basal medium label (Avoid covering the basal medium lot # and expiration date) to avoid confusion or possible double supplementation.
5. Record the new expiration date on the label based on the shelf life.
Note: If there is concern that sterility was compromised during the supplementation process, the entire newly prepared growth medium may be refiltered with a 0.2 μm filter to assure sterility. Routine refiltration is not recommended.
