Tightly packed monolayers will require some trituration to create one-cell suspensions. Avoid extreme trituration or shear power by resting the pipette tip against the plate edge to take care of viability. If cells usually do not dissociate nicely, incubation time with dissociation reagent might should be optimized.

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Movement cytometry can be employed to evaluate the efficiency of differentiation protocols Along with the STEMdiff™ system, by checking out specific marker expression at Just about every stage with the differentiation method. It will also be useful for other programs, together with cell sorting, immunophenotyping, and purity assessment.

Do the job promptly at this step to pool wells into a 15 mL tube. Note: For a significant-scale dissociation, make use of a multichannel pipette to pool cells into a sterile reagent reservoir. Add the pooled wells right into a 15 mL tube.

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Heading again into the plate, rinse Every nicely with 1 mL of FACS buffer and transfer the amount to your 15 mL tube. Notice: Retain cell suspension on ice immediately after transfer 지방이식 into the tube until ready to run FACS.

Observe: Proceed to antibody staining if carrying out Stay cell stem cell clinic Assessment following this phase. Alternatively, progress to fixable dye staining and PFA fixation.

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To assess the regional specificity of your smaller vs massive airway, you'll be able to carry out the following assays:

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Indeed, you’ll locate the step-by-stage protocol for TEER measurement To judge the epithelial barrier integrity in ALI cultures right here.

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