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Cells were incubated for 60 min at 37 C without CO 2 , pelleted by centrifugation for 5 min at 1200 g , washed 1x in 1 ml of ice-cold 1% BSA-DPBS, pelleted by centrifugation, resuspended in 300 l cell viability stain (1 M Calcein Violet 450 AM (CV450, eBioscience, San Diego, CA, USA) in DMEM-HI without supplements or FBS), incubated 15 min on ice for and analyzed by flow cytometry (Fortessa, BD, San Jose CA, USA) with 10 000 cellular events analyzed per sample, detecting DDAOG with 640 nm excitation and 670 nm emission, and CV450 with 405 nm excitation and 450 nm emission

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(C) Continuous monitoring performance obtained by CA, comparing MNs with and without a PU encapsulation layer