Fabrication of microfluidic systems in poly(dimethylsiloxane)

McDonald, J. Cooper; Duffy, David C.; Anderson, Janelle R.; Chiu, Daniel T.; Wu, Hongkai; Schueller, Olivier; Whitesides, George M.

doi:10.1002/(sici)1522-2683(20000101)21:1<27::aid-elps27>3.0.co;2-c

Cited by 2,904 publications

(1,005 citation statements)

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“…After the two layers were reversibly sealed, the space between microcolumns was formed as the migration region, which had same height with the microcolumns. There is no liquid leakage with the reversible sealing [20], the detailed information was shown in Fig. 1B.…”

Section: Microfluidic Device Design and Fabricationmentioning

confidence: 91%

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Characterization of the interaction between fibroblasts and tumor cells on a microfluidic co‐culture device

Liu

Qin

et al. 2010

Electrophoresis

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Fibroblasts and tumor cells have been involved in the process of cancer development, progression and therapy. Here, we present a simple microfluidic device which enables to study the interaction between fibroblasts and tumor cells by indirect contact co-culture. The device is composed of multiple cell culture chambers which are connected by a parallel of cell migration regions, and it enables to realize different types of cells to communicate each other on the single device. In this work, human embryonic lung fibroblasts cells were observed to exhibit obvious migration towards tumor cells instead of normal epithelial cells on the co-culture device. Moreover, transdifferentiation of human embryonic lung fibroblast cells was recognized by the specific expression of a-smooth musle actin, indicating the effect of tumor cells on the behavior of fibroblasts. Furthermore, multiple types of cell co-culture can be demonstrated on the single device which enables to mimic the complicated microenviroment in vivo. The device is simple and easy to operate, which enables to realize real-time observation of cell migration after external stimulus. This microfluidic device allows for the characterization of various cellular events on a single device sequentially, faciliating the better understanding of interaction between heterotypic cells in a more complex microenvironment. Keywords:Co-culture / Microfluidic / Migration / Transdifferentiation DOI 10.1002/elps.200900776 IntroductionGrowing evidences demonstrate that tumor microenvironment including stromal cells, inflammatory cells, extracellular matrix and vessels plays an important role in the development, progression and therapy of cancer [1][2][3][4][5].Fibroblasts make up the largest number of stromal cells, which are continuously influenced by soluble factors secreted by tumor cells and their migration abilities can be enhanced by tumor cells. When fibroblasts are transdifferentiated, they become activated and were termed cancerassociated fibroblasts or myofibroblasts, which will have a significant effect on the proliferation, invasion and metastasis of tumor cells [6][7][8]. Therefore, it is of great importance to investigate the heterotypic cell interactions between tumor cells and fibroblasts. At present, co-culture is often used to investigate the interactions between heterotypic cells by using Transwell assay [9,10], in which two types of cells are separated into upper and lower chambers by a semi-permeable membrane, or by bathing cells in conditioned medium [11][12][13][14]. However, these assays are limited by the tedious manual operations which require several procedures, and they are all typical end-point assays, which are difficult to realize real-time observation. In addition, they still require large consumption of cells and reagents that are expensive in amount.Microfluidics has sparked increasing interests in cellbased biological and medical analysis, due to its miniaturization, low consumption of reagents, and capabilities to integrate various experimental op...

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Section: Microfluidic Device Design and Fabricationmentioning

confidence: 91%

“…1. This device is composed of two layers, in which the top PDMS layer contained three cell culture chambers and fabricated in PDMS using a rapid prototyping technique [19,20]. Each cell culture chamber is 50 mm high, 1 mm wide and 6 mm long.…”

Section: Microfluidic Device Design and Fabricationmentioning

confidence: 99%

Characterization of the interaction between fibroblasts and tumor cells on a microfluidic co‐culture device

Liu

Qin

et al. 2010

Electrophoresis

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“…Confocal imaging of microfluidic-device-trapped islets Devices were fabricated using the elastomer polydimethylsiloxane as described [19]. Islets were labelled with 4 μmol/l Fluo-4 (Molecular Probes, Eugene, OR, USA) at room temperature for 2 h in imaging buffer (125 mmol/l NaCl, 5.7 mmol/l KCl, 2.5 mmol/l CaCl 2 , 1.2 mmol/l MgCl 2 , 10 mmol/l HEPES, 2 mmol/l glucose and 0.1% BSA, pH 7.4).…”

Section: Methodsmentioning

confidence: 99%

Hyperinsulinism in mice with heterozygous loss of KATP channels

et al. 2006

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Aims/hypothesis ATP-sensitive K + (K ATP ) channels couple glucose metabolism to insulin secretion in pancreatic beta cells. In humans, loss-of-function mutations of beta cell K ATP subunits (SUR1, encoded by the gene ABCC8, or Kir6.2, encoded by the gene KCNJ11) cause congenital hyperinsulinaemia. Mice with dominant-negative reduction of beta cell K ATP (Kir6.2[AAA]) exhibit hyperinsulinism, whereas mice with zero K ATP (Kir6.2 −/− ) show transient hyperinsulinaemia as neonates, but are glucose-intolerant as adults. Thus, we propose that partial loss of beta cell K ATP in vivo causes insulin hypersecretion, but complete absence may cause insulin secretory failure. Materials and methods Heterozygous Kir6.2 +/− and SUR1 +/− animals were generated by backcrossing from knockout animals. Glucose tolerance in intact animals was determined following i.p. loading. Glucose-stimulated insulin secretion (GSIS), islet K ATP conductance and glucose dependence of intracellular Ca 2+ were assessed in isolated islets. Results In both of the mechanistically distinct models of reduced K ATP (Kir6.2 +/− and SUR1 +/− ), K ATP density is reduced by ∼60%. While both Kir6.2 −/− and SUR1 −/− mice are glucose-intolerant and have reduced glucose-stimulated insulin secretion, heterozygous Kir6.2 +/− and SUR1 +/− mice show enhanced glucose tolerance and increased GSIS, paralleled by a left-shift in glucose dependence of intracellular Ca 2+ oscillations. Conclusions/interpretation The results confirm that incomplete loss of beta cell K ATP in vivo underlies a hyperinsulinaemic phenotype, whereas complete loss of K ATP underlies eventual secretory failure.

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“…Microfluidic devices were manufactured in glass and PDMS using standard soft lithography methods 21 with injected electrodes. Briefly, a layer of SU8 was spin coated to appropriate thickness on a silicon wafer and cured with UV light through a channel-patterned photo mask to produce a master mold.…”

Section: Manufacturing Of Chipsmentioning

confidence: 99%

“…20 These droplets can be manipulated at rates of thousands per second, using microfluidic devices manufactured by soft lithography. 21 Single cells can be encapsulated in such droplets, each droplet constituting the equivalent of a miniature test tube where each specific cell can be assayed. Microfluidic droplets can be manipulated in a multitude of ways including splitting, 22 fusion, 23 trapping, 24 injection of reagent 25 and sorting.…”

Section: Introductionmentioning

confidence: 99%

High-throughput screening for industrial enzyme production hosts by droplet microfluidics

et al. 2014

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A high-throughput method for single cell screening by microfluidic droplet sorting is applied to a whole-genome mutated yeast cell library yielding improved production hosts of secreted industrial enzymes. The sorting method is validated by enriching a yeast strain 14 times based on its α-amylase production, close to the theoretical maximum enrichment. Furthermore, a 10 5 member yeast cell library is screened yielding a clone with a more than 2-fold increase in α-amylase production. The increase in enzyme production results from an improvement of the cellular functions of the production host in contrast to previous droplet-based directed evolution that has focused on improving enzyme protein structure. In the workflow presented, enzyme producing single cells are encapsulated in 20 pL droplets with a fluorogenic reporter substrate. The coupling of a desired phenotype (secreted enzyme concentration) with the genotype (contained in the cell) inside a droplet enables selection of single cells with improved enzyme production capacity by droplet sorting. The platform has a throughput over 300 times higher than that of the current industry standard, an automated microtiter plate screening system. At the same time, reagent consumption for a screening experiment is decreased a million fold, greatly reducing the costs of evolutionary engineering of production strains.

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Fabrication of microfluidic systems in poly(dimethylsiloxane)

Cited by 2,904 publications

References 71 publications

Characterization of the interaction between fibroblasts and tumor cells on a microfluidic co‐culture device

Characterization of the interaction between fibroblasts and tumor cells on a microfluidic co‐culture device

Hyperinsulinism in mice with heterozygous loss of KATP channels

High-throughput screening for industrial enzyme production hosts by droplet microfluidics

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