Cell Recovery Considerations:
1. Read the cell manual carefully to understand the cell-related information, such as cell morphology, medium used, serum ratio, and configure the corresponding complete medium to ensure that the cell culture conditions are consistent with the instructions. If the cells are in trouble due to inconsistent culture conditions, the responsibility is the responsibility of the customer. It is recommended to resuscitate only one of the two cryotubes per cell. If there is any problem with the first recovery, please contact us in time.
2. Remove the frozen tube and immerse it in a warm water bath at 37 °C. Shake it from time to time to melt it as soon as possible. Note that the nozzle should not fall below the water surface to avoid contamination.
3. Remove the cryotube from the 37 ° C water bath, wipe the surface of the frozen tube with 75% alcohol, and place it in a clean bench. Aspirate the cell suspension, transfer to a culture flask, add 6-8 mL of culture medium, and statically incubate in a 37 ° C incubator. Change the fresh medium to remove DMSO every other day, or remove the DMSO by centrifugation and observe under a microscope. Cell state and density; for cells with slower adherence, it is recommended to remove DMSO at the time of resuscitation, and then change the cells after the cells are attached to the wall. (Primary cells: After the cell suspension was centrifuged, the cell bottom sediment was observed to determine the cell amount, and then trypan blue staining was performed to determine the cell survival rate; after the inoculation, the cell adherence amount was observed every other day to determine the cell adherence rate)
4. Adherent cells: Most cells will reattach after overnight culture, and will be passaged normally when the confluence is about 80%. If the cells still do not adhere, please use trypan blue staining to identify cell viability. If the trypan blue staining confirmed that the cell viability was normal, the cells were collected by centrifugation at 1000 rpm for 3~5 min, resuspended in fresh medium and transferred back to the culture flask for re-adherent culture; if trypan blue staining confirmed that the cells were not viable, please receive the cells. Contact us within 24 hours and give feedback on real cell status photos and trypan blue stained photos, we will help you out as soon as possible.
5. Suspended cells: After overnight culture, most of the cells will gradually rejuvenate, shiny and full. If the cell gloss is dim, trypan blue staining is used to identify the cell viability. If trypan blue staining confirms that the cells are not viable, please contact us within 24 hours after receiving the cells, and feed back the real cell status photos and trypan blue stained photos. We will help you out as soon as possible.
6. It is recommended that the customer take a few photos of the cells every day after receiving the cells, record the cell status, and communicate with the technical department of the Chinese microbial strain inquiry network.
Shanghai Enjosim Medical Technology Co., Ltd, Jiangsu Enjosim Medical Technology Co., Ltd , https://www.enjosimmedical.com