Multichannel Electrosonic Actuation Microarray for Cell-Based Screening
Department of Health and Human Services
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Small Business Information
OPENCELL TECHNOLOGIES, LLC
OPENCELL TECHNOLOGIES, LLC, 500 Bishop St NE, Ste E3, ATLANTA, GA, 30318
Socially and Economically Disadvantaged:
AbstractDESCRIPTION (provided by applicant): High-throughput, parallel delivery and transfection of biologically relevant material into cells remains a difficult task. The proposed work addresses this issue through integration of multiple Electrosonic Actuation Microarrays (EAMs) into a single MIST (Multiple Integrated Sample Treatment) device that is optimized for parallel drug and/or gene/nucleic acid delivery and transfection into several cell samples via precise control of biophysical action (i.e., concurrent sono/mechanoporation and electroporation). The device format is compatible with existing large-scale cell-handling equipment. The EAM is a microelectromechanical systems (MEMS)-enabled device that ejects a sample containing biological cells through microscopic (of order size of a single cell) nozzles with incorporated electroporation electrodes, thereby opening pores in the cell membrane via combined mechano/electroporation for uptake of nanomaterials. The high ultrasonic frequency of operation and array format enable fast processing of large cell populations (up to millions of cells per second). A single reservoir system can accommodate a wide range of prescribed volumes, from ~100 nL to arbitrarily large sample sizes. By arraying EAM together, MIST enables parallel and uniform treatment of several different samples simultaneously. Economical fabrication enables fluid handling components of the device to be made disposable, which eliminates cross-sample contamination. MIST is suited to basic/applied research, as well as diagnostic and therapeutic uses. Design, development and evaluation of cell treatment by MIST will build on experience gained through characterization of a multimode bench-top-scale prototype, STEAM (Single-sample Treatment via Electrosonic Actuation Microarray). A growing understanding of the bioeffects induced in certain cell types during EAM treatment will facilitate the transition to a multichannel device. MIST development has the potential to greatly accelerate discovery of cancer therapies by enabling simultaneous screening of synthetic small interfering RNA (siRNA) for different effects on cell function. Demonstrating the feasibility of our approach will open opportunities for commercialization of MIST in other markets. The primary objective of this Phase I SBIR is to expand our single-sample STEAM platform into the multi-sample MIST device and to demonstrate parallel delivery and transfection of multiple biomolecules into different cell samples. To achieve this objective, (1) a MIST platform that is optimized for cell treatment will be developed, and (2) safe handling with regard to viability and proliferation, uniformity of treatment, and the transfection capabilities of MIST will be evaluated using standard cell assays, confocal fluorescence microscopy and flow cytometry. In addition, MIST treatment will be compared with conventional lipofectamine-mediated and electroporation-based transfection. PUBLIC HEALTH RELEVANCE: Development of a MIST (Multiple Integrated Sample Treatment) platform will address the current need for high-throughput, parallel delivery and transfection of biologically relevant material into cells. Further, because the device format is compatible with existing large-scale cell handling equipment (e.g., 384-well plate-based systems), it has the potential to accelerate discovery of potential cancer therapies through efficient screening of large complementary DNA (cDNA) and small interfering RNA (siRNA) libraries. Simultaneous transfection of multiple different DNA plasmids encoding fluorescent proteins using MIST, will demonstrate its feasibility in a wide range of drug discovery and gene therapy applications.
* information listed above is at the time of submission.