Fabrication and use of a transient contractional flow device to quantify the sensitivity of mammalian and insect cells to hydrodynamic forces

A microfluidic device was fabricated via photolithographic techniques which can create transient elongational and shear forces ranging over three orders of magnitude while still maintaining laminar flow conditions. The contractional fluid flow inside the microfluidic device was simulated with FLUENT (a computational fluid dynamics computer program) and the local deformation forces were characterized with the scalar quantity, local energy dissipation rate. The sensitivities of four cell lines (CHO, HB‐24, Sf‐9, and MCF7) were tested in the device. The results indicate that all four cell lines are able to withstand relatively intense energy dissipation rates (up to 104–105 kW/m3), which is orders of magnitude higher than the maximum local energy dissipation rates generated by impellers in bioreactors, but comparable to that associated with small bursting bubbles. While the concept that suspended animal cells are relatively robust with respect to purely hydrodynamic forces in bioprocess equipment is well known, these results quantitatively demonstrate these observations. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 80: 428–437, 2002.

[1]  J. Birch,et al.  Reactor design for large scale suspension animal cell culture , 1999, Cytotechnology.

[2]  K. Schügerl,et al.  Response of mammalian cells to shear stress , 1991, Applied Microbiology and Biotechnology.

[3]  Sadettin S. Ozturk,et al.  Engineering challenges in high density cell culture systems , 2004, Cytotechnology.

[4]  C. R. Thomas,et al.  Prediction of mechanical damage to animal cells in turbulence , 2004, Cytotechnology.

[5]  Alvin W. Nienow,et al.  Homogenisation and oxygen transfer rates in large agitated and sparged animal cell bioreactors: Some implications for growth and production , 2004, Cytotechnology.

[6]  Analysis of void removal in liquid composite molding using microflow models , 2002 .

[7]  Jian Zhang,et al.  Polymerization optimization of SU-8 photoresist and its applications in microfluidic systems and MEMS , 2001 .

[8]  J. Chalmers,et al.  Cell damage of microcarrier cultures as a function of local energy dissipation created by a rapid extensional flow. , 2000, Biotechnology and bioengineering.

[9]  B. Loechel Thick-layer resists for surface micromachining , 2000 .

[10]  Christian Trägårdh,et al.  Scale-up of Rushton turbine-agitated tanks , 1999 .

[11]  E. Papoutsakis,et al.  Increased agitation intensity increases CD13 receptor surface content and mRNA levels, and alters the metabolism of HL60 cells cultured in stirred tank bioreactors. , 1998, Biotechnology and bioengineering.

[12]  Suzanne M. Kresta,et al.  Turbulence in stirred tanks: Anisotropic, approximate, and applied , 1998 .

[13]  Dependence of the quality of thick resist structures on resist baking , 1998 .

[14]  J. Chalmers,et al.  Study of hydrodynamics in microcarrier culture spinner vessels: A particle tracking velocimetry approach , 2000, Biotechnology and bioengineering.

[15]  S L Diamond,et al.  Fluid shear stress induction of the transcriptional activator c‐fos in human and bovine endothelial cells, HeLa, and Chinese hamster ovary cells , 2000, Biotechnology and bioengineering.

[16]  J. Frangos,et al.  Fluid flow rapidly activates G proteins in human endothelial cells. Involvement of G proteins in mechanochemical signal transduction. , 1996, Circulation research.

[17]  S. Kresta,et al.  Impact of tank geometry on the maximum turbulence energy dissipation rate for impellers , 1996 .

[18]  M. Al‐Rubeai,et al.  Death mechanisms of animal cells in conditions of intensive agitation , 1995, Biotechnology and bioengineering.

[19]  M. Gray,et al.  The effects of impeller and tank geometry of circulatory time distributions in stirred tanks , 1995 .

[20]  Carl M. Stoots,et al.  Mean velocity field relative to a Rushton turbine blade , 1995 .

[21]  Jeffrey J. Chalmers,et al.  Computer simulations of the rupture of a gas bubble at a gas—liquid interface and its implications in animal cell damage , 1994 .

[22]  M Al-Rubeai,et al.  Estimation of disruption of animal cells by turbulent capillary flow , 1993, Biotechnology and bioengineering.

[23]  John R. Blake,et al.  Gas bubbles bursting at a free surface , 1993, Journal of Fluid Mechanics.

[24]  A. Nienow,et al.  Further studies of the culture of mouse hybridomas in an agitated bioreactor with and without continuous sparging. , 1992, Journal of biotechnology.

[25]  L V McIntire,et al.  Hydrodynamic shear stress and mass transport modulation of endothelial cell metabolism , 1991, Biotechnology and bioengineering.

[26]  Gary B. Tatterson,et al.  Fluid mixing and gas dispersion in agitated tanks , 1991 .

[27]  E. Papoutsakis,et al.  Damage mechanisms of suspended animal cells in agitated bioreactors with and without bubble entrainment , 1990, Biotechnology and bioengineering.

[28]  E. Papoutsakis,et al.  The protective effect of serum against hydrodynamic damage of hybridoma cells in agitated and surface-aerated bioreactors. , 1990, Journal of biotechnology.

[29]  Alvin W. Nienow,et al.  The effects of agitation intensity with and without continuous sparging on the growth and antibody production of hybridoma cells , 1989 .

[30]  L V McIntire,et al.  Fluid flow stimulates tissue plasminogen activator secretion by cultured human endothelial cells. , 1989, Science.

[31]  M S Croughan,et al.  Growth and death in overagitated microcarrier cell cultures , 1989, Biotechnology and bioengineering.

[32]  L. McIntire,et al.  Shear Stress Effects on Human T Cell Function , 1988 .

[33]  J. Couderc,et al.  Study by laser Doppler anemometry of the turbulent flow induced by a Rushton turbine in a stirred tank: Influence of the size of the units—II. Spectral analysis and scales of turbulence , 1988 .

[34]  Michael Yianneskis,et al.  An experimental study of the steady and unsteady flow characteristics of stirred reactors , 1987, Journal of Fluid Mechanics.

[35]  幸道 岡本,et al.  攪拌槽内のエネルギー消散速度分布とその液-液分散および固-液物質移動への影響 , 1979 .

[36]  K. Van't Riet,et al.  The trailing vortex system produced by Rushton turbine agitators , 1975 .

[37]  A J Sinskey,et al.  Effect of shear on the death of two strains of mammalian tissue cells , 1971, Biotechnology and bioengineering.

[38]  L. A. Cutter Flow and turbulence in a stirred tank , 1966 .

[39]  R. Aris Vectors, Tensors and the Basic Equations of Fluid Mechanics , 1962 .