TY - JOUR
T1 - Microfluidic bioreactors for culture of non-adherent cells
AU - Shah, Pranjul Jaykumar
AU - Vedarethinam, Indumathi
AU - Kwasny, Dorota
AU - Andresen, Lars
AU - Dimaki, Maria
AU - Skov, Søren
AU - Svendsen, Winnie Edith
PY - 2011/8
Y1 - 2011/8
N2 - Microfluidic bioreactors (μBR) are becoming increasingly popular for cell culture, sample preparation and analysis in case of routine genetic and clinical diagnostics. We present a novel μBR for non-adherent cells designed to mimic in vivo perfusion of cells based on diffusion of media through a sandwiched membrane. The culture chamber and perfusion chamber are separated by a sandwiched membrane and each chamber has separate inlet/outlets for easy loading/unloading of cells and perfusion of the media. The perfusion of media and exchange of nutrients occur through the sandwiched membrane, which was also verified with simulations. Finally, we present the application of this device for cytogenetic sample preparation, whereby we culture and arrest peripheral T-lymphocytes in metaphase and later fix them in the μBR. The expansion of T-lymphocytes from an unknown patient sample was quantified by means of CFSE staining and subsequent counting in a flow cytometer. To conclude on the applicability of μBR for genetic diagnostics, we prepare chromosome spreads on glass slides from the cultured samples, which is the primary step for metaphase FISH analysis.
AB - Microfluidic bioreactors (μBR) are becoming increasingly popular for cell culture, sample preparation and analysis in case of routine genetic and clinical diagnostics. We present a novel μBR for non-adherent cells designed to mimic in vivo perfusion of cells based on diffusion of media through a sandwiched membrane. The culture chamber and perfusion chamber are separated by a sandwiched membrane and each chamber has separate inlet/outlets for easy loading/unloading of cells and perfusion of the media. The perfusion of media and exchange of nutrients occur through the sandwiched membrane, which was also verified with simulations. Finally, we present the application of this device for cytogenetic sample preparation, whereby we culture and arrest peripheral T-lymphocytes in metaphase and later fix them in the μBR. The expansion of T-lymphocytes from an unknown patient sample was quantified by means of CFSE staining and subsequent counting in a flow cytometer. To conclude on the applicability of μBR for genetic diagnostics, we prepare chromosome spreads on glass slides from the cultured samples, which is the primary step for metaphase FISH analysis.
UR - http://www.scopus.com/inward/record.url?scp=79958146709&partnerID=8YFLogxK
U2 - 10.1016/j.snb.2011.02.021
DO - 10.1016/j.snb.2011.02.021
M3 - Journal article
AN - SCOPUS:79958146709
SN - 0925-4005
VL - 156
SP - 1002
EP - 1008
JO - Sensors and Actuators B: Chemical
JF - Sensors and Actuators B: Chemical
IS - 2
ER -