Do you do have a protocol or proposed antibodies to conduct ICC staining on my fully differentiated ALI cultures?
After the run is total, retrieve the Shipping and delivery Cartridge from the instrument. The cell sample must be at The underside or facet of the collection tube. Observe: It is usually recommended to spin down the Supply Cartridge inside a mini-centrifuge for any few seconds for full volume recovery.
The in vitro differentiation of human pluripotent stem cells (hPSCs) into unique cell and tissue varieties allows the research of human biology without the need to have for primary tissues or in vivo designs.
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Set the “unstained” tube aside. To the “viability dye” sample, centrifuge the tube at 300 x g
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TEER measurements may be executed continuously, without the need of triggering damage to the cell lifestyle. You are able to carry out a weekly TEER stem cell clinic time program to explain the barrier function all over the whole process of ALI tradition differentiation.
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Note: For a really confluent tradition, cultures may be a bit far more clumpy through dissociation. To beat this, Carefully scrape the monolayer that has a pipette idea right after introducing the dissociation reagent to aid the dissociation throughout incubation.
Sure, you’ll discover the step-by-stage protocol for TEER measurement To guage the epithelial barrier integrity in ALI cultures in this article.
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Immediately thaw cells within a 37°C h2o tub by Carefully shaking the cryovial. Take away the vial when a small frozen cell pellet stays. Note: It is necessary to work speedily in the following methods to make sure significant cell viability and recovery.