Gibco细胞冻存液12648-010

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产品名称：细胞冻存液

英文名称：Recovery™ Cell Culture Freezing Medium

品牌：Gibco

规格：50ML

货号：12648-010

储存温度：-20度— -5度

有效期：12个月Description

RecoveryTM Cell Culture Freezing Medium is a complete ready-to-use cryopreservation medium with proven performance on a broad spectrum of mammalian cell lines. RecoveryTM Cell Culture Freezing Medium is a proprietary formulation based on Dulbecco’s Modified Eagle Medium (High Glucose) with optimized levels of fetal bovine serum, bovine serum and DMSO (10%) providing improved viability and cell recovery after thawing.Product use

For Research Use Only. Not for use in diagnostic procedures.

Safety information

Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves.

Cryopreservation

For optimum results, cells should be in mid-log phase of growth with >90% viability at the time of freezing. Similar protocols may be substituted.

1. Thaw RecoveryTM Cell Culture Freezing Medium, mix well and keep at 2°C to 8°C until use.

2. For suspension cells proceed to step 3. For adherent cells, gently detach cells from the substrate on which they are growing using a suitable dissociation reagent such as TrypLETM. Resuspend cells in complete medium required for that cell type.

3. Transfer cell suspension to a sterile 15-mL centrifuge tube.

4. Determine the viable cell density and percent viability using

a Countess® Automated Cell Counter (similar automated or manual methods may be used) and calculate the required volume of RecoveryTM Cell Culture Freezing Medium to give a final cell density of 1 × 106 to 1 × 107 cells/mL.

5. Centrifuge cell suspension at 100–200 × g for 5–10 minutes. Aseptically decant supernatant without disturbing the cell pellet.

Note: Centrifugation speed and duration may vary depending on cell type.

6. Resuspend the cell pellet in (2°C to 8°C) chilled RecoveryTM Cell Culture Freezing Medium at recommended viable cell density for specific cell type (typically 1 × 106 cells/mL or greater) .

7. Dispense aliquots of cell suspension (mix frequently to maintain a homogeneous cell suspension) into cryovials according to the manufacturer’s specifications (i.e., 1.5 mL in a 2-mL cryovial).

8. Achieve cryopreservation in an automated or manual controlled rate freezing apparatus following standard procedures (approximay 1 ?C decrease per minute).

9. Transfer frozen cells to liquid nitrogen, (vapor phase) storage at –200°C to –125°C is recommended.Recovery

1. Remove cells from cryo-storage and rapidly thaw

(<1 minute) frozen vial in a 37°C water bath until only a small amount of ice remains.

2. Transfer cell suspension to a sterile 15-mL conical tube. Add, dropwise, the appropriate pre-warmed complete growth medium to a total volume of 10 mL. Ensure complete mixing with regular gentle swirling.

3. Centrifuge cell suspension at 100–200 × g for 5–10 minutes. Note: Centrifugation speed and duration may vary depending on cell type.

4. Ascertain presence of cell pellet. Aseptically decant supernatant without disturbing the cell pellet.

5. Gently resuspend cell pellet in an appropriate volume

(e.g., 5 mL per 25 cm2 surface area) of pre-warmed complete growth medium.

6. Transfer cell suspension to sterile culture vessel and place into the recommended culture environment.

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