Organoid Harvesting Solution

This Organoid Harvesting Solution enables gentle and rapid recovery of cells cultured on basement membrane matrix, while maintaining cell viability throughout the process. It is optimized for efficient organoid retrieval prior to passaging, cryopreservation, or downstream analysis.

(1) Observe organoid morphology under microscope; proceed with passaging when organoid density reaches 500 organoids per well or individual organoid diameter is 100-200 μm, to maintain optimal organoid health and proliferation rate. (2) Prepare all required reagents and consumables in advance. (3) In a biosafety cabinet, aspirate old culture medium from the plate, add pre-cooled Organoid Washing Solution (GBOR0004) for one wash. Change pipette tip, add Organoid Harvesting Solution (500 μL/well for 24-well plate), and incubate at room temperature for 5 minutes. (4) Using a 1000 μL pipette tip pre-rinsed with Rinse Solution (GBOR0006), dislodge organoids by pipetting or scraping. Collect into a 15 mL centrifuge tube, add 5 mL Organoid Washing Solution, pipette organoids 5-10 times, place on ice for 10 minutes, then centrifuge at 200×g for 5 minutes at 2-8°C. Discard supernatant. (5) Choose mechanical or enzymatic dissociation method based on organoid developmental stage and morphology. (6) Centrifuge at 200×g for 5 minutes at 2-8°C, collect cell pellet, remove as much supernatant as possible, then add matrix gel or Organoid Cryopreservation Medium for subsequent expansion culture or cryopreservation.