A microfluidic array with cellular valving for single cell co-culture

Frimat, Jean-Philippe; Becker, Marco; Chiang, Ya-Yu; Marggraf, Ulrich; Janasek, Dirk; Hengstler, Jan G.; Franzke, Joachim; West, Jonathan

doi:10.1039/c0lc00172d

Cited by 168 publications

(159 citation statements)

References 30 publications

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“…Unlike our original co-culture system which required cellular valving (i.e. adhesion and flattening) before the addition of a second cell set, 22 the microfluidic neuron arraying circuit has a central channel which permits the simultaneous arraying of neurons in both flanking culture compartments. An alternative circuit was also developed which uses the same differential flow microarraying principle.…”

Section: Microfluidic Neuron Arrayingmentioning

confidence: 99%

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Microfluidic construction of minimalistic neuronal co-cultures

et al. 2013

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In this paper we present compartmentalized neuron arraying (CNA) microfluidic circuits for the preparation of neuronal networks using minimal cellular inputs (10-100-fold less than existing systems).The approach combines the benefits of microfluidics for precision single cell handling with biomaterial patterning for the long term maintenance of neuronal arrangements. A differential flow principle was used for cell metering and loading along linear arrays. An innovative water masking technique was developed for the inclusion of aligned biomaterial patterns within the microfluidic environment. For patterning primary neurons the technique involved the use of meniscus-pinning micropillars to align a water mask for plasma stencilling a poly-amine coating. The approach was extended for patterning the human SH-SY5Y neuroblastoma cell line using a poly(ethylene glycol) (PEG) back-fill and for dopaminergic LUHMES neuronal precursors by the further addition of a fibronectin coating. The patterning efficiency E patt was .75% during lengthy in chip culture, with y85% of the outgrowth channels occupied by neurites. Neurons were also cultured in next generation circuits which enable neurite guidance into all outgrowth channels for the formation of extensive inter-compartment networks. Fluidic isolation protocols were developed for the rapid and sustained treatment of the different cellular and sub-cellular compartments. In summary, this research demonstrates widely applicable microfluidic methods for the construction of compartmentalized brain models with single cell precision. These minimalistic ex vivo tissue constructs pave the way for high throughput experimentation to gain deeper insights into pathological processes such as Alzheimer and Parkinson Diseases, as well as neuronal development and function in health.

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Section: Microfluidic Neuron Arrayingmentioning

confidence: 99%

“…cellular valving). 22 In a further design iteration, the central channel can also serve as another culture compartment for other neurons, oligodendrocytes and other heterotypic combinations (ESI, 3 Fig. 1(B)).…”

Section: In Chip Patterned Culture Of Neuronal Networkmentioning

confidence: 99%

Microfluidic construction of minimalistic neuronal co-cultures

et al. 2013

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“…Such approaches, which allowed for trapping single cells by actuating integrated valve systems 31 or exploiting physical obstacles as traps 14 , have a drawback in terms of device fabrication. Recently a new approach was presented to successfully isolate single cells within small trapping elements 15,19 . While this method was based on trap configurations geometrically optimized for achieving high trapping efficiency, it resulted not suitable for adherent cell cultures, lacking in sufficient space for cell spreading and subsequent proliferation.…”

Section: Discussionmentioning

confidence: 99%

“…However, these approaches for trapping single cells require complex and expensive fabrication procedures due to the need of multilayered devices 13,14 . As an alternative, small trapping units were recently proposed [15][16][17][18][19] to block cells along their fluidic path; however their integration with chambers for subsequent culture is not trivial. An interesting approach allowing for trapping and culturing single cells within a microfluidic devices based on a variable resistance mechanism -the cells functioning as fluid stopper once trapped-at the outlet of a culture chamber was recently reported by Hong and colleagues 9 .…”

Section: Introductionmentioning

confidence: 99%

High-throughput microfluidic platform for adherent single cells non-viral gene delivery

et al. 2015

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The widespreading of gene therapy as therapeutic tool relies on the development of DNAcarrying vehicles devoid of safety concerns. Opposite to viral vectors, non-viral gene carriers are promising in this perspective, although their low transfection efficiency leads to the necessity of further optimizations. Aiming at overcoming the limitation of traditional macroscale approaches, mainly consisting in time-consuming and simplified models, a microfluidic strategy was developed for transfection studies on single cells in a highthroughput and deterministic fashion. A single cell trapping mechanism was implemented based on dynamic variation of fluidic resistances. At this purpose, we conceived a roundshaped culture chamber integrated with a bottom trapping junction which modulates the hydraulic resistance. Several layouts of the chamber were designed and computationally validated for the optimization of the single cell trapping efficacy. The optimized chamber layout was integrated in a polydimethylsiloxane (PDMS) microfluidic platform presenting two main functionalities: (i) 288 chambers for trapping single cells, (ii) a serial dilution generator provided with chaotic mixing properties, able to deliver to chambers both soluble factors and non-diffusive particles (i.e. polymer/DNA complexes, polyplexes) under spatio-temporally controlled chemical patterns. Devices were experimentally validated and allowed for trapping individual human glioblastoma-astrocytoma epithelial-like cells (U87-MG) with a trapping efficacy of about 40%. Cells were cultured within the device and underwent preliminary transfection experiments using 25kDa linear polyethylenimine (lPEI)-based polyplexes, confirming the potentiality of the proposed platform for the future high-throughput screening of gene delivery vectors and the optimization of transfection protocols.

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“…This technique has been used by many other researchers and has been shown to be reliable. 15,16 However, while higher capturing flow rates do improve trapping efficiency, capture still remains a probabilistic phenomenon.…”

Section: Introductionmentioning

confidence: 99%

Streamline based design guideline for deterministic microfluidic hydrodynamic single cell traps

et al. 2015

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A prerequisite for single cell study is the capture and isolation of individual cells. In microfluidic devices, cell capture is often achieved by means of trapping. While many microfluidic trapping techniques exist, hydrodynamic methods are particularly attractive due to their simplicity and scalability. However, current design guidelines for single cell hydrodynamic traps predominantly rely on flow resistance manipulation or qualitative streamline analysis without considering the target particle size. This lack of quantitative design criteria from first principles often leads to non-optimal probabilistic trapping. In this work, we describe an analytical design guideline for deterministic single cell hydrodynamic trapping through the optimization of streamline distributions under laminar flow with cell size as a key parameter. Using this guideline, we demonstrate an example design which can achieve 100% capture efficiency for a given particle size. Finite element modelling was used to determine the design parameters necessary for optimal trapping. The simulation results were subsequently confirmed with on-chip microbead and white blood cell trapping experiments. V C 2015 AIP Publishing LLC.

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A microfluidic array with cellular valving for single cell co-culture

Cited by 168 publications

References 30 publications

Microfluidic construction of minimalistic neuronal co-cultures

Microfluidic construction of minimalistic neuronal co-cultures

High-throughput microfluidic platform for adherent single cells non-viral gene delivery

Streamline based design guideline for deterministic microfluidic hydrodynamic single cell traps

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