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Mix gently before transferring cells to the conical tube that contains 10 mL of pre-warmed wash medium. Observe: Wash the pipette suggestion and cryovial While using the clean medium at the least three instances to help enhance Restoration.
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See information exhibiting outstanding ALI tradition morphology and higher epithelial cell marker expression applying these recommended Transwell® inserts >
Established the “unstained” tube aside. For your “viability dye” sample, centrifuge the tube at 300 x g
Assess the standing by examining a droplet and halting the dissociation when all over 지방이식 eighty% with the cell suspension is solitary-celled. Prevent about-digestion.
Based on the quantity of mucus accumulation, a second clean could also be necessary. See how a mucus wash is performed In this particular ALI lifestyle differentiation movie (skip to 02:24) >
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Perform cell counts employing an automatic cell counter. Take note: Reduced viability is often optimized by the length of collagenase or ACCUTASE™ incubation. It can be optimized 지방흡입 by reducing the volume of triturations.
This really is an conclude-issue protocol designed to dissociate PSC-derived kidney organoids into just one-cell suspension and fixation for downstream movement cytometry Examination.
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