Figure 1
From: Development of a miniaturized 96-Transwell air–liquid interface human small airway epithelial model
Semi-automated hSAE cell seeding and automated long-term maintenance of ALI cultures in 96-Transwell plates, a process overview. (a) hSAE cells from a frozen stock were thawed and resuspended in expansion medium, kept in suspension by gentle shaking of the falcon, and seeded into 96-Transwell plates using a Multidrop Combi device. Enabling of long-term maintenance of hSAE cells under ALI conditions in 96-Transwell plates via an integrated robotic system. Cell culture room was equipped with an HEPA-filtered air feed and ambient air overpressure. For plate maintenance, an automated microplate incubator (1) was combined with a laminar flow hood (3) via an airtight lock (2). A plate shuttle system (4) transported single 96-Transwell plates out of the incubator through the airtight lock into the laminar flow hood. Plates were collected from the airtight lock through a robotic arm (5). Inside the laminar flow hood a combined washer dispenser device (6) exchanged the medium from the Transwell plates, which was connected to a reservoir bag (7). Regular medium exchange on each plate was controlled and scheduled by a specialized software (8). (b) Representative overview of the customized constructed transepithelial electrical resistance (TEER) device. PC-controlled high-throughput TEER measurement for epithelial monolayer, including a data acquisition tool (1). The main components include: the robotic sampler, containing a panel of 96 individual electrodes, which moves the electrodes in each well of the Transwell plate (2), a base plate for the 96-well tray, including an electrode washing station (3), which is connected to different valves for waste, ethanol and sterile water. (c) Magnification of one single electrode pair, lowered in to the upper and basal compartment of the Transwell plate (Details of employed software see Supplementary Information, online). (d) Cross section of one single Transwell during a classical TEER measurement. (e) Overview of the actual device placed under the laminar flow hood. (f) Magnification of the robotic sampler, containing a panel of 96 individual (dual) electrodes (g) Magnification of the 96 individual electrode pairs per Transwell insert.
