Abstract
Flow-based microfluidic systems have been widely utilized for cell migration studies given their ability to generate versatile and precisely defined chemical gradients and to permit direct visualization of migrating cells. Nonetheless, the general need for bulky peripherals such as mechanical pumps and tubing and the complicated setup procedures significantly limit the widespread use of these microfluidic systems for cell migration studies. Here we present a simple method to power microfluidic devices for chemotaxis assays using the commercially available ALZET® osmotic pumps. Specifically, we developed a standalone chemotaxis platform that has the same footprint as a multiwell plate and can generate well-defined, stable chemical gradients continuously for up to 7 days. Using this platform, we validated the short-term (24 hours) and long-term (72 hours) concentration dependent PDGF-BB chemotaxis response of human bone marrow derived mesenchymal stem cells.
Highlights
While cell migration is critical for embryogenesis, homeostasis, and tissue regeneration, a simple, robust, and accessible in vitro assay for the study of chemotaxis that can generate long-term stable linear gradients from nearly any agent has remained elusive
Despite substantial improvements made by flow-based microfluidic gradient generators, traditional chemotaxis assays remain the method of choice for studying cell migration due to their simplicity [3]
For long-term studies (.12 hours), this system is impractical as it is limited by the small capacity of the osmotic pump (120 mL) which only lasts for several hours at the rate of 20 mL/hr
Summary
While cell migration is critical for embryogenesis, homeostasis, and tissue regeneration, a simple, robust, and accessible in vitro assay for the study of chemotaxis that can generate long-term stable linear gradients from nearly any agent has remained elusive. Flow-based systems are relatively complex to utilize (compared to placing transwell inserts in a multiwell plate) and often require bulky equipment such as external electrical or pneumatic pumps, tubing, and fittings.[2] Assembling these components is laborious and they cannot be integrated into common cell culture procedures [7]. This becomes problematic for long-term chemotaxis studies where the experimental setup must be maintained under standard cell culture conditions for days (i.e. at 37uC and 5% CO2). The need to fabricate osmotic pumps discourages the routine use of this system
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
