Organoids refer to tissue analogues with a certain spatial structure formed by three-dimensional cultivation of adult stem cells or pluripotent stem cells in vitro. Organs are highly consistent with their source tissues in terms of tissue structure, cell type, self-renewal ability, and function, thus demonstrating unique advantages in multiple biomedical fields such as developmental biology, disease modeling, precision medicine, drug research and development, gene and cell therapy, infection and immunity, and regenerative medicine.
Product Description:
Organoid Dissociation Solution (ODS) can be applied to the routine passage of various mammalian derived organs, such as humans, mice, pigs, bats, cows, etc. It can separate organoids from matrix gel and gently digest them into small cell clusters or single cells, while maintaining their growth vitality after passage.
Product instructions
Digestive passage of organoids1. Add 5-10 times the volume of organoid matrix gel mixture to the recovered organoid organs, known as Organoid Dissociation Solution, After blowing and mixing, incubate at 37 ° C for 1-8 minutes to dissociate the organoids (it is necessary to preheat the required volume of digestive solution at 37 ° C in advance, and the digestion time for single-layer structured organoids is usually 0.5-3 minutes, while for multi-layer or larger organoids, the digestion time is 3-8 minutes).
Attention: During this operation, the digestion process must be carefully monitored to avoid excessive digestion. During the digestion process, a pipette can be used to blow and mix to aid digestion. A small amount of digestive suspension can also be taken in real-time and observed under a microscope for digestion. When a large number of single cells or a diameter of 50 are observed μ Digestion is considered complete when the cell clusters below m are present.
2. Add at least five times the volume of organoid basic culture medium to the suspension that has been confirmed to have completed digestion for dilution, thereby terminating digestion.
Note: After a longer digestion process, fetal bovine serum (FBS) can be added appropriately to the final concentration of 2% -5% to ensure the vitality of the digested cells.
3. Centrifuge the organoid suspension obtained in the previous step (horizontal centrifuge rotor, 150-300g, 3 minutes), discard the supernatant, and add the basic culture medium again to resuspend the organoid sediment.
4. Centrifuge the organoid suspension obtained in the previous step (horizontal centrifuge rotor, 150-300g, 3 minutes). After discarding the supernatant, the organoid obtained can be used for subsequent organoid culture, cryopreservation, and other experimental operations.