STEM-CELLBANKER® DMSO Free GMP grade is a chemically defined freezing solution that does not contain DMSO as an anti-freezing agent. It was developed for customers who do not prefer to use DMSO-containing cryopreservation solution due to a variety of their intended uses. STEM-CELLBANKER® DMSO Free GMP grade is also manufactured in compliance with JPN, EU, US, and PIC/S GMP guidelines.
When tested in the following cells with this product, 80-90% of viability of cells was observed.
Complete List of Cells Tested [PDF]
|129SV||Mouse ES cell|
|201B7||Human iPS cell|
|Caco-2||Human colonic adenocarcinoma cell|
|Canine PBMC||Canine peripheral blood mononuclear cells|
|Feline PBMC||Feline peripheral blood mononuclear cells|
|HeLa||Human uterine cervical carcinoma cell|
|HepG2||Human hepatocellular carcinoma cell|
|Jurkat||Human T-cell line|
|K562||Human Caucasian chronic myelogenous leukemia cell|
|NBM-Lu||Normal newborn murine fibroblast cell line|
|P3/x63-Ag8.U1||Murine myeloma cell|
|SK007||Human B-cell line|
|UE6E7-16||Human Mesenchymal cell|
|UE7T-13||Human Mesenchymal Stem cell|
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 : 20mL & 100mL
- Storage and Stability : 2 to 8℃ or below -20℃
- Expiration date : 3 years after the date of manufacture
20ml size x 4
References & Literature
Ueda, H. et al. Establishment of in vitro 3D spheroid cell cultivation from human gynecologic cancer tissues. STAR Protocols 2, 100354 (2021) doi: 10.1016/j.xpro.2021.100354.
Sako, K. et al. Optimization of Spheroid Colony Culture and Cryopreservation of Nucleus Pulposus Cells for the Development of Intervertebral Disc Regenerative Therapeutics. Applied Sciences 11, (2021) doi: 10.3390/app11083309.
Yagishita, S. et al. Characterization of the large-scale Japanese patient-derived xenograft (J-PDX) library. Cancer Science 112, 2454–2466 (2021) doi: 10.1111/cas.14899.
Mikłosz, A. et al. Does TBC1D4 (AS160) or TBC1D1 Deficiency Affect the Expression of Fatty Acid Handling Proteins in the Adipocytes Differentiated from Human Adipose-Derived Mesenchymal Stem Cells (ADMSCs) Obtained from Subcutaneous and Visceral Fat Depots? Cells 10, (2021) doi: 10.3390/cells10061515.
Ueno, K. et al. Freezing of cell sheets using a 3D freezer produces high cell viability after thawing. Biochemistry and Biophysics Reports 28, 101169 (2021) doi: 10.1016/j.bbrep.2021.101169.
FOR RESEARCH USE ONLY, NOT FOR USE IN DIAGNOSTIC PROCEDURES