Dielectrophoretic cell capture
Inventors
Gaitan, Michael • Elliott, John T. • Gordon, Jennifer Hong • Reyes-Hernandez, Darwin R. • Dittrich, Petra S. • Hanke, Conni
Assignees
National Institute of Standards and Technology NIST
Publication Number
US-9101939-B2
Publication Date
2015-08-11
Expiration Date
2032-09-21
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Abstract
Various aspects are described for selectivity capturing cells or bioparticles on designated surfaces in dielectrophoretic systems and processes. A particular adhesive composition is described for enhancing cell retention. In addition, certain permeable polyester membranes used in the systems and processes are also described.
Core Innovation
The present invention addresses difficulties associated with selectively capturing and retaining cells or bioparticles on designated surfaces during dielectrophoresis (DEP). It introduces a layered composition including at least one adhesion material layer and a polycation material layer, where the latter provides an exposed face for retaining bioparticles during DEP. The invention also provides dielectric assemblies comprising a polyester permeable membrane with electrically conductive members and the layered adhesive composition disposed on the membrane and/or conductive members.
A primary problem solved is the challenge of retaining cells trapped by DEP in a fluid flow field once DEP forces are removed. Existing approaches such as polyelectrolyte multilayers (PEMs) enable short-term cell anchorage but exhibit deleterious effects on cells after 24 hours, failing to maintain long-term cell viability and function post DEP trapping. The invention provides a hybrid cell adhesive material (hCAM) comprising fibronectin and polycation layers atop PEMs, enabling instantaneous cell adhesion after DEP trapping and supporting long-term cell proliferation and differentiation, demonstrating cell function retention over days in microfluidic systems.
Claims Coverage
The claims include two independent claims and associated dependent claims, covering both a layered assembly for DEP and a method for retaining cells and bioparticles during DEP.
Layered assembly for DEP with polyester permeable membrane and adhesive layers
An assembly comprising a polyester permeable membrane defining an outer face, at least one electrically conductive member disposed on the membrane's outer face, and a layered composition on the membrane and/or conductive member, the composition including at least one adhesion material layer and a polycation material layer disposed on the adhesion layer.
Method for retaining cells and bioparticles during DEP on a treated surface
A method comprising providing a DEP system generating a non-uniform electric field proximate a target surface including a polyester permeable membrane with electrically conductive members, applying a layered composition including at least one adhesion material layer topped with a polycation material layer having an exposed face for retaining cells and bioparticles, and performing DEP to cause cells and bioparticles to contact and be retained on the exposed face of the composition.
The claims collectively cover the structure of layered assemblies combining a porous polyester membrane, conductive electrodes, and specific layered adhesion and polycation materials to enhance selective capture and retention of cells or bioparticles during DEP, as well as methods of applying these layered compositions and performing DEP to achieve retention of cells and bioparticles.
Stated Advantages
The hCAM allows instantaneous cell anchorage immediately after DEP trapping, overcoming detachment issues when DEP forces are removed.
The layered adhesive composition supports long-term cell viability, proliferation, and differentiation, demonstrated for over 8 days post DEP trapping, enabling extended in vitro cell experiments.
The use of permeable polyester membranes with patterned conductive microelectrodes maintains membrane permeability, hydrophilicity, and surface roughness, preserving functional properties necessary for cell culture and DEP.
The integrated system allows for precise cell positioning and retention under continuous fluid flow fields without disturbing channel flow, thereby providing a tool suitable for studying cell-cell interactions and co-cultures.
Documented Applications
In vitro long-term cell experiments preserving cell function such as proliferation, motility, and differentiation under DEP manipulation.
Microfluidic devices for selective capture, patterning, and retention of cells and bioparticles using DEP forces combined with adhesive layered surfaces.
Multilayer microfluidic platforms with permeable polyester membranes and DEP microelectrodes to study cell-cell communication, co-culture systems, controlled microenvironments, and drug transport studies.
Cell differentiation studies, exemplified by P19 cell differentiation into neuron-like cells after DEP trapping on the hybrid adhesive material.
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