Dynamic cell culture describes the in vitro culture of cells in the presence of applied mechanical stress. Of great emerging interest is the ability to control the flow of fluid, such as medium, over cells while they grow. Flow rate can be generated very precisely using microfluidic instruments and is an ideal way to better replicate the dynamic environment that cells normally reside in. Medium microfluidic perfusion mimics the flow of blood, allowing nutrient exchange and the removal of waste products, and also adds shear stress to cells.
In this application note we describe how to perfuse cells for dynamic cell culture. Perfuse with the automated OB1 pressure controller using a variety of flow profiles, including constant flow or intermittent, steady rate or pulsatile, or create a customised profile, e.g. to mimic the cardiac rhythm. Read also our application notes about automated cell seeding and cell staining for dynamic cell culture.
Some main applications of dynamic cell culture include:
Cell and biology pack
Microfluidic chamber designed for perfusion (IBIDI µSlide I Luer)
Flow controller OB1 Mk3+
Mux-distributor
Microfluidic bubble trap
Flow sensor
Manifold 9 ports
Tubing, fittings and reservoirs
Follow our guide to seed and adhere cells in a microfluidic chip.
TIP: To ensure solid attachment, leave cells 12-18h after seeding before starting flow
Perfuse: Switch flow to reservoir containing medium 1.
TIP: use a step gradient to slowly increase flow rate to desired when scheduling continuous perfusion
Switch medium: If desired, switch flow to reservoir containing medium 2.
Flow profiles can be steady, pulsatile or custom, e.g. mimic the cardiac rhythm
ESI software
HeLa cell attachment and proliferation (t=2h, 18h, 26h). Perfusion (intermittent at 20 µL/min for 10 min per hour) was started at t=18h.
Application note written by Lisa MUIZNIEKS, Subia BANO, Camila BETTERELLI GIULIANO and Jessica AYACHE.
Acknowledgement: This work was done thanks to the funding of European Union’s Horizon 2020 research and innovation programme (PANBioRA project, grant agreement No 760921; MECH-LoC project, MSCA grant agreement No 793749; MTOAC project, MSCA grant agreement No 795754; Protomet project, MSCA grant agreement No 813873).
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