hPSC Gentle Dissociation Solution is a gentle and efficient cell dissociation solution launched by Aladdin, specifically designed for pluripotent stem cells. This product is a gentle digestive enzyme that enables single-cell passaging, ensuring minimal cell damage during dissociation and improving cell survival rate after passaging. The digestion time is 3–5 minutes, which simplifies the cell passaging process. This product is of animal-free origin, and its production process strictly follows the ISO9001 system and complies with GMP guidelines, ensuring high quality and safety.
Product Features
1.Gentle and efficient, with high post-passaging survival rate.
2.Enables single-cell passaging.
Experimental Preparation
1.Coating: Use Matrigel to coat the culture dish 1 hour in advance at 37°C.
2.Reagent preparation: Take the hPSC Gentle Dissociation Solution and the prepared medium out of the refrigerator half an hour in advance, protect from light, and equilibrate to room temperature; pre-incubate other reagents such as PBS at 37°C.
Operating Method (using a 10 cm dish as an example)
1. Check cells: Observe cell status under a microscope to ensure that the cell morphology shows colonies with smooth edges, and that the confluence is appropriate (approximately 70–75%), meeting the passaging criteria.
2.Washing: Aspirate the old medium, add 10 mL PBS to rinse the cells, avoiding direct streaming onto the cells to prevent detachment.
3. Digestion: Add 5 mL of pre-warmed hPSC Gentle Dissociation Solution, and place the cells in a 37°C incubator for 3–5 minutes of digestion.
4. Collection: Confirm under a microscope that the cells have dissociated into small clumps and have detached from the culture plate. Gently pipette the cells using PBS, transfer the cell suspension to a centrifuge tube, and rinse the culture plate again with PBS to maximize cell collection.
5.Preparation of single cells (optional): Pipette 6–10 times using a pipette to ensure that 80–90% of the cells become single cells.
Note: If single-cell passaging is required, this step can be selected.
6.Centrifugation: Centrifuge at room temperature, 1200 rpm for 3 minutes.
7.Resuspension: Carefully aspirate the supernatant, add 10 mL of medium, and gently resuspend the cells 4–5 times with a pipette, avoiding bubble formation.
8.Seeding: Remove the Matrigel-coated culture dish from the 37°C incubator, aspirate the coating solution, add 10 mL of medium, inoculate the cell suspension into the plate at the recommended cell density, and check the seeding density under a microscope to ensure it is appropriate.
9.Mixing: Gently rock the culture dish back and forth and left to right to ensure even cell distribution. After overnight incubation, change the medium the next day, and replace the medium every 24 hours until the cells are ready for the next operation.
