Product Information

• Application
• Protocol

Description:

STEM-CELLBANKER^® EX GMP grade (SCB EX) is a cell cryopreservation solution that expands the possibilities of cell therapy. SCB EX is a simple composition and composed of ingredients that are proven in intravenous injections. It is a DMSO-containing cryopreservation solution and a Xeno-Free product formulated serum free and animal component free. SCB EX can also preserve human hematopoietic stem cells and human adipose-derived stromal/stem cells. It is completely free of serum and animal derived component, and contains only USP, EP, JP graded ingredients or JP excipients. SCB EX is manufactured in a facility compliant with JP, EU, US and PIC/S GMP guidelines and the production and the quality are controlled in compliance with JP GMP guidelines.

※This product has been registered in MF within PMDA in 2020 and BBMF within CBER (Center for Biological Evaluation and Research)/FDA (the U.S. Food and Drug Administration) in 2022.

※The composition of this product is the same as HSC-BANKER^® GMP grade.

Application:

Cryopreservation performance test using Adipose-Derived Stromal/Stem Cells (ADSC)

Test：EMO Biomedicine

1. Cell Viability Test

ADSCs were cryopreserved with SCB EX for 4, 12, 24 and 48 weeks. The cell viability was evaluated immediately after thawing (left panel) and overnight after thawing (right panel). The cells cryopreserved with SCB EX showed high viability even after long-term cryopreservation and maintained their high viability even after overnight resting.

2. Cell Recovery Test

ADSCs were cryopreserved with SCB EX for 4, 12, 24 and 48 weeks. And then, we evaluated the cell attachment overnight after thawing (Cell Recovery; left panel) and the cell growth rate (Doubling Time; right panel) after thawing and 72 hours of cultivation. The results showed that even after long-term cryopreservation, the cell recovery and the cell growth rate was not affected.

3. Cell Identity Test

ADSCs were cryopreserved with SCB EX for 4, 12, 24 and 48 weeks. And then, we evaluated the cell identity after thawing using positive and negative markers with flow cytometry. The results showed that even after long-term cryopreservation, the percentage of the cell labeled with each markers was not affected.

Positive Marker; CD29, CD73, CD90 and CD105
Negative Marker; CD11b, CD19, CD34, CD45 and HLA-DR

Data of Positive Marker

4. Cellular function Test
　(IDO Quantification and Suppression of T Cell Proliferation Assay)

ADSCs were cryopreserved with SCB EX for 4, 12, 24 and 48 weeks. And then, we evaluated the cellular functions of ADSCs after thawing. The results showed that even after long-term cryopreservation, the IDO expression and the suppression rate of T cell proliferation in the presence of MSCs were not affected.

※Indoleamine-2,3-dioxygenase (IDO)

Protocol（CELLBANKER^® series）

Freezing

For optimum results, cells for cryopreservation should be in log phase of growth. Similar or standard freezing protocols may be substituted.

1. Examine and make sure the cell culture free of contamination, in healthy situation and proper confluency, etc.
2. Perform a cell count to determine the viability of cells
3. Gently pellet the cells centrifugation (3-5 minutes at 1,000~2,000rpm, 4℃). Remove the supernatant using a aspirator.
4. Gently suspend the cells with CELLBANKER cryopreservation medium (1 ml for 5×10⁵ - 5×10⁶ cells).
5. Dispense the cell suspension in 1ml aliquots to cryopreservation vials that have been labeled with the cell line name, cell concentration, passage date and other essential information.
6. Place the vials directly in a -80℃ for storage. If necessary, transfer the frozen vials to a liquid nitrogen storage tank after the vials have been frozen for at least 24 hours.

IMPORTANT: Optimum protocol may change with the cell types.

Thawing

1. Remove the frozen cell from storage and quickly thaw in a 37 ℃ shaking water bath.
2. Immediately dilute and gently mix each 1ml of cells with 10ml of complete cell culture medium.
3. Gently pellet the cells centrifugation (3-5 minutes at 1,000~2,000rpm, 4 ℃). Remove the supernatant aspirator.
4. Gently suspend the cells with appropriate volume of complete cell culture medium. And plate in a culture flask.
5. Continue the further culture procedures according to standard protocols.

※If the survival rate is low, try slow freezing method using a freezing box, among other things.

Storage and Stability:

Store at 2 to 8 or below -20 degrees Celsius. In unopened condition and under storage temperatures (2-8℃ ), the product will be stable for 3 years after the date of manufacture.

SDS : STEM-CELLBANKER^® EX GMP grade

Disclaimer
STEM-CELLBANKER^® EX GMP grade is not itself a pharmaceutical. Therefore, ZENOAQ and ZENOAQ RESOURCE make no warranty, express or implied, as to the fitness and suitability of this product for any particular purpose and/or merchantability unless the use is intended for research.