Do you do have a protocol or proposed antibodies to conduct ICC staining on my fully differentiated ALI cultures?
Mix Carefully in advance of transferring cells into a conical tube containing 10 mL of pre-warmed clean medium. Take note: Clean the pipette suggestion and cryovial Along with the clean medium at the very least three periods to help you boost Restoration.
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Alternatively, dissociate organoids working with Gentle Cell Dissociation Reagent (GCDR) with no prior retrieval of organoids within the Matrigel® dome. This can be unquestionably quicker but bears the chance of inadequate cell dissociation as a consequence 지방흡입 of challenges with Matrigel® and will be more tense into the cells.
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Here is an index of published literature that cites culturing of nasal epithelial cells making use of PneumaCult™ culture media:
In case cell clumping is observed immediately after thawing cryopreserved cells, it is recommended to filter aggregated suspensions by way of a 37 µm cell strainer (e.g. Catalog #27250) for exceptional outcomes. In such a case, the run time within the CellPore™ Transfection System may well need to be improved to ten seconds.
To evaluate the regional specificity on the modest vs huge airway, you may perform the subsequent assays:
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