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 is Xeno-free, serum-free, and animal component-free formulation. SCB EX can also preserve human hematopoietic stem cells and human adipose-derived stromal/stem cells.
SCB EX is manufactured in compliance with JPN, EU, US, and PIC/S GMP guidelines.
Application: Cryopreservation performance test using Adipose-Derived Stromal/Stem Cells (SDSC)
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.
Cell Recovery Test
ADSCs were cryopreserved with SCB EX for 4, 12, 24 and 48 weeks. Then 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.
Cell Identity Test
ADSCs were cryopreserved with SCB EX for 4, 12, 24 and 48 weeks. Then 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 marker was not affected.
Positive Marker; CD29, CD73, CD90 and CD105
Negative Marker; CD11b, CD19, CD34, CD45 and HLA-DR
Data of Positive Marker
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. Then 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.
|Adipose-Derived Stromal/Stem Cells (ADSC)||STEM-CELLBANKER® EX GMP grade||EMO Biomedicine|
|Cord blood derived hematopoietic stem cells||HSC-BANKER® GMP grade||The Japan society for Hematopoietic cell transplantation (2015)|
For optimum results, cells for cryopreservation should be in log phase of growth. Similar or other standard freezing protocols may be substituted.
- Examine and make sure the cell culture is free of contamination, in healthy and at proper confluency.
- Perform a cell count to determine the viability of cells.
- Centrifuge at 1,000 – 2,000 rpm, 4°C for 3 to 5 minutes to gently pellet the cells. Remove the supernatant with an aspirator.
- Gently suspend STEM-CELLBANKER® GMP grade cryopreservation medium (1 mL for 5×10⁵ – 5×10⁶ cells).
- Transfer 1 mL of the cell suspension to cryopreservation vial labeled with appropriate information (the cell line name, concentration, passage date etc.).
- Place the vials directly in -80℃ for storage.
- (OPTIONAL) 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.
- Remove the cryopreservation vial from the freezer and quickly thaw cells in a 37°C shaking water bath or shake by hand.
- Transfer the content to a centrifugation tube then immediately dilute and gently mix with 10mL of complete cell culture medium. Using CELLOTION® instead of complete culture medium will prevent adhesion of cells to the wall of the tube, increasing the recovery rate.
- Centrifuge cells at 1,000 – 2,000 rpm, 4°C for 3 to 5 minutes. Remove the supernatant with an aspirator.
- Gently resuspend the cells with an appropriate volume of complete cell culture medium then plate in a culture flask or plate
- Continue the culture procedures according to standard protocols.
- Manufactured By : Zenogen Pharma Co., Ltd
- Size : 100mL
- Storage and Stability : 2 to 8℃ or below -20℃
- Expiration date : 3 years after the date of manufacture
- Disclaimer: STEM-CELLBANKER® EX GMP grade is not itself a pharmaceutical. Therefore, 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.
References & Literature
Mizui, T., Chen, H., Lee, S. & Fukui, M. Study on cryopreservation of human adipose-derived stromal cells. Cytotherapy 23, S60–S61 (2021) doi: 10.1016/S1465324921003546.
FOR RESEARCH USE ONLY, NOT FOR USE IN DIAGNOSTIC PROCEDURES