Organoid Cryopreservation Medium (Serum Free) 

MCE Organoid Cryopreservation Medium (Serum Free) is a ready-to-use cryopreservation medium, widely applicable to organoids and cell lines derived from various mammals (such as human, mouse, pig, cow, etc.).

This product is serum-free and contains no animal-derived components, reducing the risk of contamination by bacteria, viruses, mycoplasma, and other pathogens, ensuring the safe cryopreservation of organoids.

4℃, 3 years

It is recommended to select organoids/cells in good growth condition for cryopreservation.

1. Preparation of Cell Freezing Container: Pre-warm the cell freezing container to room temperature or 4°C in advance.

2. Organoid/Cell Collection: Add an appropriate amount of organoid basal medium to the organoid culture plate. Use a cell scraper or a 1 mL pipette tip to detach the contents of the culture wells and transfer them to a centrifuge tube. Centrifuge at 150 - 300 g for 3 min, discard the supernatant, and collect the organoids.

3. Organoid Aliquoting: After centrifugation, add pre-chilled organoid cryopreservation solution to the organoids/cells, adjust the cell concentration to 1 × 10³ - 1 × 10⁷ cells or the equivalent number of organoids per mL, mix thoroughly, and transfer to cryovials.

Note: a. It is recommended to use 1 × 10³ - 1 × 10⁷ cells/mL organoid cryopreservation medium.

b. The cell suspension volume in the cryovial should be greater than 0.5 mL.

c. For organoids with large cryopreservation volumes, multi-layered epithelium, or squamous cell structures, due to their dense structure, it is recommended to digest the organoids using Organoid Dissociation Solution (MCE Catalog No: HY-K6013) appropriately. After digestion, wash the organoids twice with organoid basal culture medium before aliquoting and cryopreserving.

4. Organoid Cryopreservation: Transfer the cryovials to the cell freezing container, and then place the freezing container into a -80°C freezer. After 12 - 24 h, transfer to liquid nitrogen for long-term storage.

Pre-warm the complete organoid/cell basal culture medium to 37°C before starting the experiment.

1. Rapid Thawing: Quickly thaw the cryovial in a 37°C water bath. When only a small amount of ice remains in the vial, immediately remove the vial from the water bath and transfer it to the sterile working area.

2. Dilution: Transfer the cryopreservation suspension to a centrifuge tube. Slowly add 5-10 times the volume of pre-warmed basal culture medium and gently mix.

3. Centrifugation: Centrifuge the organoid/cell suspension at 150 - 300 g for 3 min, discard the supernatant and resuspend the organoid/cell pellet in basal culture medium. Repeat centrifugation once to wash the cells.

4. Preparation for Culturing: Resuspend the obtained organoid/cell pellet in basal culture medium for subsequent organoid/cell culture.

1. This product is sterile and should be handled using aseptic techniques. It is recommended to aliquot and store to avoid contamination.

2. This product contains 10% DMSO.

3. This product is for R&D use only, not for drug, household, or other uses.

4. For your safety and health, please wear a lab coat and disposable gloves to operate.