Figure 1.

Generation of the single cell clone “Arlo.” A) Schematic depicting the single‐cell printing procedure (detailed in B) and subsequent passaging strategy for the single‐cell clone “Arlo” originating from a polyclonal hAELVi suspension that demonstrated TEER values >1000 Ω cm² in the previous passage. B) Before single‐cell printing, the cell diameter of the polyclonal hAELVi cell suspension was determined via a Casy cell counter to define the printing parameters. Single cells within the printing parameters (bordered green) were confirmed by an image‐based algorithm and then deposited into a single well of a 96‐well plate. Cells that did not meet the printing criteria were discarded via vacuum aspiration after ejection from the printing nozzle (bordered purple or red). C,D) Light microscopic images showing morphological differences between C) the polyclonal hAELVi cell line and D) the single cell clone “Arlo” when cultured in T25 cm² culture flasks for 7 d. Scale: 50 µm for all images displayed. Panel A) was partly generated using Servier Medical Art, provided by Servier, licensed under a Creative Commons Attribution 3.0 unported license.