Vitroscope AS*

Faktry, Leirfossvegen 5D, 7037 Trondheim, Norway. *Correspondence:

[email protected]

Introduction

Here we describe a step-by-step process for perfusing adherent HEK293 cells for 24 hours in vitro.alive (Vitroscope AS). vitro.alive is a sensor integrated cell culture system designed to provide microenvironmental control in 2D and 3D flow-based assays.

vitro.alive’s 2D chamber is compatible with adherent cell lines, and the following procedure can be taken into consideration when planning other vitro.alive experiments.

Materials and Reagents

• HEK293 cells
• Cell culture medium: Dulbecco’s Modified Eagle’s Medium - high glucose, Fetal Bovine Serum (10%) and Pen/Strep (1%) (Sigma-Aldrich)
• Double distilled water ($ddH_2O$)
• Ethanol (75%) (antibac)
• Trypsin-EDTA (25%) (gibico)
• PBS wash buffer
• Trypan Blue (0.4%) (ThermoFisher)

Equipment

• alive (Vitroscope)$^1$$^)$
  • 1x adapter
  • 1x chamber
    • 1x locking pin
    • 1x silicone O-ring
    • 1x glass slide (76 mm x 38 mm x 1 mm) $^2$$^)$
    • 1x silicone filling tube
  • 1x power connector
  • 1x pin pusher