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Mix gently just before transferring cells to your conical tube made up of 10 mL of pre-warmed wash medium. Notice: Wash the pipette suggestion and cryovial Along with the clean medium at least three times to assist improve recovery.
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It can be crucial to quench the dissociation reagent making use of FACS buffer or stop medium, by including at the least the exact same or double the quantity of the dissociation reagent.
Cross-portion histology accompanied by hematoxylin and eosin (H&E) staining to assess the thickness with the small or big airway epithelium
For greatest effects, the total quantity of cargo extra mustn't exceed 10% with the response volume. Decreasing the response quantity to below 80 µL may bring about decrease modifying efficiencies and cell recoveries.
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To assess the regional specificity from the stem cell clinic small vs substantial airway, you can accomplish the subsequent assays:
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Certainly, you’ll locate the stage-by-action protocol for TEER measurement to evaluate the epithelial barrier integrity 가슴수술 in ALI cultures listed here.
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Rapidly thaw cells inside of a 37°C drinking water bath by Carefully shaking the cryovial. Eliminate the vial when a little frozen cell pellet remains. Notice: It is necessary to work speedily in the following techniques to make sure significant cell viability and recovery.