Biological implications of polydimethylsiloxane-based microfluidic cell culture.
Polydimethylsiloxane (PDMS) has become a staple of the microfluidics community by virtue of its simple fabrication process and material attributes, such as gas permeability, optical transparency, and flexibility. As microfluidic systems are put toward biological problems and increasingly utilized as cell culture platforms, the material properties of PDMS must be considered in a biological context. Two properties of PDMS were addressed in this study: the leaching of uncured oligomers from the polymer network into microchannel media, and the absorption of small, hydrophobic molecules (i.e. estrogen) from serum-containing media into the polymer bulk. Uncured PDMS oligomers were detectable via MALDI-MS in microchannel media both before and after Soxhlet extraction of PDMS devices in ethanol. Additionally, PDMS oligomers were identified in the plasma membranes of NMuMG cells cultured in PDMS microchannels for 24 hours. Cells cultured in extracted microchannels also contained a detectable amount of uncured PDMS. It was shown that MCF-7 cells seeded directly on PDMS inserts were responsive to hydrophilic prolactin but not hydrophobic estrogen, reflecting its specificity for absorbing small, hydrophobic molecules; and the presence of PDMS floating in wells significantly reduced cellular response to estrogen in a serum-dependent manner. Quantification of estrogen via ELISA revealed that microchannel estrogen partitioned rapidly into the surrounding PDMS to a ratio of approximately 9:1. Pretreatments such as blocking with serum or pre-absorbing estrogen for 24 hours did not affect estrogen loss from PDMS-based microchannels. These findings highlight the importance of careful consideration of culture system properties when determining an appropriate environment for biological experiments.
Regehr
Keil J
KJ
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53706-1609, USA.
Domenech
Maribella
M
Koepsel
Justin T
JT
Carver
Kristopher C
KC
Ellison-Zelski
Stephanie J
SJ
Murphy
William L
WL
Schuler
Linda A
LA
Alarid
Elaine T
ET
Beebe
David J
DJ
D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods / Q000592:standards; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry; D004129:Dimethylpolysiloxanes / Q000032:analysis / Q000737:chemistry / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D004967:Estrogens / Q000737:chemistry; D006801:Humans; D008156:Luciferases / Q000235:genetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism
D003470:Culture Media; D004129:Dimethylpolysiloxanes; D004967:Estrogens; D018808:Transcription Factor AP-1; C013830:baysilon; D008156:Luciferases