Shear-induced intracellular loading of cells with molecules by controlled microfluidics.
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 microm 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.
Hallow
Daniel M
DM
School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, 311 Ferst Drive, Atlanta, Georgia 30332, USA.
Seeger
Richard A
RA
Kamaev
Pavel P
PP
Prado
Gustavo R
GR
LaPlaca
Michelle C
MC
Prausnitz
Mark R
MR
D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017022:Flow Injection Analysis / Q000295:instrumentation* / Q000379:methods; D005452:Fluoresceins / Q000493:pharmacokinetics*; D006801:Humans; D008297:Male; D040542:Mechanotransduction, Cellular; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods; D011471:Prostatic Neoplasms / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000493:pharmacokinetics*; D033081:Shear Strength
D005452:Fluoresceins; D012710:Serum Albumin, Bovine; C007740:fluorexon