Generation of Stable Concentration Gradients in 2D and 3D Environments Using a Microfluidic Ladder Chamber
Our microfluidic Ladder Chamber is capable of generating stable concentration gradients across 2D surfaces and 3D matrices. Neutrophil chemotaxis was successfully demonstrated in the Ladder Chamber. Compared to other assays, this chamber offers simplicity and improved stability of the gradients. In addition, the chamber provides a high-throughput approach for rapid characterization of chemotactic responses. The diffusion chamber also provides a straightforward method for generating concentration gradients in three-dimensional gels. This can be adapted to a variety of gels, allowing chemotaxis to be studied in different 3D environments. The simple nature of this approach, coupled with its versatility, will make it broadly applicable in cell migration research.
In our paper entitled 'Generation Of Stable Concentration Gradients In 2D And 3D Environments Using A Microfluidic Ladder Chamber' currently located at http://www.springerlink.com/content/u621226324l10743/fulltext.pdf, we delineate a simple microfluidic device for generating stable concentration gradients in 2D and 3D environments. The device, termed the Ladder Chamber, uses a two-compartment diffusion system to generate steady state gradients across flow-free channels that connect the source and sink channels. To demonstrate the utility of the Ladder Chamber for cell migration, neutrophil chemotaxis was successfully observed in soluble chemoattractant (IL-8) gradient. The Ladder Chamber's simple design and experimental implementation make it an attractive approach for investigating cell migration and other biological experiments in well-defined gradients in 2D surfaces as well as in 3D gels.A microfluidics-based method for generating stable concentration gradients with controlled profiles has been developed (Dertinger et al. 2001; Jeon et al. 2000). However, the microfluidic chemotaxis chamber has two main drawbacks. First, since the chamber depends on diffusion across laminar streams to establish gradients, the cells are exposed to constant shear flow, which can influence directional motility. In addition, most of the autocrine/paracrine factors that are secreted by the cells will be washed away. Second, as in most conventional chemotaxis assays, migration in the microfluidic chemotaxis chamber is limited to two-dimensional (2D) substrates. Two-dimensional cell migration assays do not address in-vivo processes that involve migration in a three-dimensional (3D) matrix, such as neutrophil chemotaxis after transmigration, and cancer cell invasion.
Patent Number: US7947491B2
Application Number: US2007849194A
Inventor: Jeon, Noo Li | Saadi, Wajeeh | Rhee, Seog Woo | Mosadegh, Bobak | Huang, Carlos
Priority Date: 31 Aug 2006
Priority Number: US7947491B2
Application Date: 31 Aug 2007
Publication Date: 24 May 2011
IPC Current: C12M000300 | C12M000134
US Class: 4352885
Assignee Applicant: The Regents of the University of California
Title: Microfluidic gradient devices
Usefulness: Microfluidic gradient devices
Summary: As gradient device used in a system and platform for generating a gradient (claimed), for studying live cells; used to successfully observe chemotaxis of a cell in a gradient of a soluble factor such as, for e.g. chemokine; used to examine the behavior of cells in a controlled fluidic microenvironment containing autocrine/paracrine factors for cell signaling investigations.
Novelty: Gradient device for generating concentration gradients comprises gradient chamber associated with source and sink channel, configured to facilitate formation of gradient generating region exclusively by diffusion
生物医学
医疗装置
7947491
Tech ID/UC Case 18794/2006-169-0 Related Cases 2006-169-0
美国
