D002-Dissociation Solution Ⅱ

Dissociation SolutionⅡ

Product Description

This product is applicable to the dissociation of various mouse organoids and can also be used for the dissociation of individual tissues. It features high purity and good specificity, and can avoid the potential damage to cells caused by other enzymes existing in some trypsin extracts.

Storage

Store at 2-8℃ for 12 months.

Instructions for Use

(1) Take the Dissociation Solution II out of the 2-8°C refrigerator and place it at room temperature for 10 minutes for preheating before use.

(2) Use a 1,000 μL pipette tip that has been rinsed with the rinsing solution (B006-5) to draw an appropriate amount of the Dissociation Solution II and add it to the centrifuge tube for collecting organoids (Table 1). Resuspend the organoids and transfer the organoid suspension to a 24-well plate.

(3) Place the 24-well plate in a 37°C incubator with 5% CO₂ and let it stand for digestion for about 5 - 15 minutes. Every 5 minutes, use a 1,000 μL pipette tip rinsed with the rinsing solution to gently pipette the organoids (5 - 10 times). Observe the degree of enzymatic hydrolysis under the microscope. When most of the cells in the field of view are single cells and small cell clusters, the enzymatic hydrolysis can be terminated (Figure 1). If the enzymatic hydrolysis time is continued to be increased, the proportion of single cells will increase and the cell viability will decrease. It is not recommended to over-digest the organoids into single cells.

Figure 1. Dissociation of organoids by Dissociation Solution II. 

Under the microscope, it was observed that as the enzymatic hydrolysis time increased, the marginal cells of the organoids gradually detached, and the organoids lost their intact morphology until they were completely dissociated. After pipetting with a 1000 μL pipette tip, when mostly single cells and small cell clusters were observed under the microscope, the digestion could be terminated (the degree of digestion within the red frame). Scale bar: 100 μm.

Table 1. Requirement Table for the Amount of Enzymatic Digestion Solution II Added

Precautions

This product is strictly reserved for scientific research and shall not be applied to clinical diagnosis, treatment, food, or pharmaceuticals. Additionally, it is prohibited from being stored in residential premises.