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Shear-induced intracellular loading of cells with molecules by controlled microfluidics

✍ Scribed by Daniel M. Hallow; Richard A. Seeger; Pavel P. Kamaev; Gustavo R. Prado; Michelle C. LaPlaca; Mark R. Prausnitz


Book ID
101726916
Publisher
John Wiley and Sons
Year
2008
Tongue
English
Weight
356 KB
Volume
99
Category
Article
ISSN
0006-3592

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✦ Synopsis


Abstract

This study tested the hypothesis that controlled flow through microchannels can cause shear‐induced intracellular loading of cells with molecules. The overall goal was to design a simple device to expose cells to fluid shear stress and thereby increase plasma membrane permeability. DU145 prostate cancer cells were exposed to fluid shear stress in the presence of fluorescent cell‐impermeant molecules by using a cone‐and‐plate shearing device or high‐velocity flow through microchannels. Using a syringe pump, cell suspensions were flowed through microchannels of 50–300 µm diameter drilled through Mylar® sheets using an excimer laser. As quantified by flow cytometry, intracellular uptake and loss of viability correlated with the average shear stress. Optimal results were observed when exposing the cells to high shear stress for short durations in conical channels, which yielded uptake to over one‐third of cells while maintaining viability at approximately 80%. This method was capable of loading cells with molecules including calcein (0.62 kDa), large molecule weight dextrans (150–2,000 kDa), and bovine serum albumin (66 kDa). These results supported the hypothesis that shear‐induced intracellular uptake could be generated by flow of cell suspensions through microchannels and further led to the design of a simple, inexpensive, and effective device to deliver molecules into cells. Such a device could benefit biological research and the biotechnology industry. Biotechnol. Bioeng. 2008;99: 846–854. © 2007 Wiley Periodicals, Inc.


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