Microfluidic sorting of microtissues.

Increasingly, invitro culture of adherent cell types utilizes three-dimensional (3D) scaffolds or aggregate culture strategies to mimic tissue-like, microenvironmental conditions. In parallel, new flow cytometry-based technologies are emerging to accurately analyze the composition and function of these microtissues (i.e., large particles) in a non-invasive and high-throughput way. Lacking, however, is an accessible platform that can be used to effectively sort or purify large particles based on analysis parameters. Here we describe a microfluidic-based, electromechanical approach to sort large particles. Specifically, sheath-less asymmetric curving channels were employed to separate and hydrodynamically focus particles to be analyzed and subsequently sorted. This design was developed and characterized based on wall shear stress, tortuosity of the flow path, vorticity of the fluid in the channel, sorting efficiency and enrichment ratio. The large particle sorting device was capable of purifying fluorescently labelled embryoid bodies (EBs) from unlabelled EBs with an efficiency of 87.3% ± 13.5%, and enrichment ratio of 12.2 ± 8.4 (n = 8), while preserving cell viability, differentiation potential, and long-term function.

[1]  H. Sollinger,et al.  Validation of Large Particle Flow Cytometry for the Analysis and Sorting of Intact Pancreatic Islets , 2005, Transplantation.

[2]  Hong Wu,et al.  Continuous sorting of heterogeneous-sized embryoid bodies. , 2010, Lab on a chip.

[3]  J. Crook,et al.  Attachment and growth of human embryonic stem cells on microcarriers. , 2008, Journal of biotechnology.

[4]  J. Squier,et al.  Particle size limits when using optical trapping and deflection of particles for sorting using diode laser bars. , 2009, Optics express.

[5]  Tony J Collins,et al.  ImageJ for microscopy. , 2007, BioTechniques.

[6]  Jeffrey S Erickson,et al.  Two simple and rugged designs for creating microfluidic sheath flow. , 2008, Lab on a chip.

[7]  David J. Mooney,et al.  Microenvironmental regulation of biomacromolecular therapies , 2007, Nature Reviews Drug Discovery.

[8]  D. Joseph,et al.  Nonlinear mechanics of fluidization of beds of spherical particles , 1987, Journal of Fluid Mechanics.

[9]  H Morgan,et al.  On-chip high-speed sorting of micron-sized particles for high-throughput analysis. , 2005, IEE proceedings. Nanobiotechnology.

[10]  Sangeeta N Bhatia,et al.  Multiplexed, high-throughput analysis of 3D microtissue suspensions. , 2010, Integrative biology : quantitative biosciences from nano to macro.

[11]  J. Bailey,et al.  Flow effects on the viability and lysis of suspended mammalian cells , 1987, Biotechnology Letters.

[12]  Wee Keong Nah,et al.  The osteogenic differentiation of adipose tissue-derived precursor cells in a 3D scaffold/matrix environment. , 2008, Current drug discovery technologies.

[13]  S. Takayama,et al.  Gravity-driven microfluidic particle sorting device with hydrodynamic separation amplification. , 2007, Analytical chemistry.

[14]  R. Tompkins,et al.  Continuous inertial focusing, ordering, and separation of particles in microchannels , 2007, Proceedings of the National Academy of Sciences.

[15]  R. Nerem,et al.  Altered response of vascular smooth muscle cells to exogenous biochemical stimulation in two- and three-dimensional culture. , 2003, Experimental cell research.

[16]  Harm A Nieuwstadt,et al.  Microfluidic particle sorting utilizing inertial lift force , 2011, Biomedical microdevices.

[17]  Stefan Johansson,et al.  On-chip fluorescence-activated cell sorting by an integrated miniaturized ultrasonic transducer. , 2009, Analytical chemistry.

[18]  K. Anseth,et al.  Small functional groups for controlled differentiation of hydrogel-encapsulated human mesenchymal stem cells. , 2008, Nature materials.

[19]  Buddy D Ratner,et al.  Biomaterials: where we have been and where we are going. , 2004, Annual review of biomedical engineering.

[20]  R. Stovel,et al.  The influence of particles on jet breakoff. , 1977, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[21]  Jin-Ming Lin,et al.  Particle sorting using a porous membrane in a microfluidic device. , 2011, Lab on a chip.

[22]  Ross A. Marklein,et al.  Homogeneous and organized differentiation within embryoid bodies induced by microsphere-mediated delivery of small molecules. , 2009, Biomaterials.

[23]  Mina J Bissell,et al.  Modeling tissue-specific signaling and organ function in three dimensions , 2003, Journal of Cell Science.

[24]  Sangeeta N Bhatia,et al.  Engineering tissues for in vitro applications. , 2006, Current opinion in biotechnology.

[25]  H. Amini,et al.  Label-free cell separation and sorting in microfluidic systems , 2010, Analytical and bioanalytical chemistry.

[26]  Brenda M Ogle,et al.  Multiphoton Flow Cytometry to Assess Intrinsic and Extrinsic Fluorescence in Cellular Aggregates: Applications to Stem Cells , 2010, Microscopy and Microanalysis.