A High-Throughput Microfluidic Platform for Mammalian Cell Transfection and Culturing

Woodruff, Kristina; Maerkl, Sebastian J.

doi:10.1038/srep23937

Cited by 41 publications

(39 citation statements)

References 50 publications

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“…To address these challenges, significant efforts have been employed integrating miniaturized cell culture systems with high-throughput liquid handling automation. 10 These systems are usually coupled with microliter scale high throughput analytics to quickly identify potential candidate cell lines for manufacturing. 11 For example, a common miniaturization platform is to use shaken microtiter plates coupled with automated liquid handling.…”

Section: Introductionmentioning

confidence: 99%

A novel mammalian cell line development platform utilizing nanofluidics and optoelectro positioning technology

Tan

Gupta

et al. 2018

Biotechnology Progress

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Generating a highly productive cell line is resource intensive and typically involves long timelines because of the need to screen large numbers of candidates in protein production studies. This has led to miniaturization and automation strategies to allow for reductions in resources and higher throughput. Current approaches rely on the use of standard cell culture vessels and bulky liquid handling equipment. New nanofludic technologies offer novel solutions to surpass these limits, further miniaturizing cell culture volumes (10 times smaller) by growing cells on custom nanofluidic chips. Berkeley Lights' OptoElectro Positioning technology projects light patterns to activate photoconductors that gently repel cells to manipulate single cells on nanofluidic culturing chips. Using a fully integrated technology platform (Beacon), common cell culture tasks can be programmed through software, allowing maintenance and analysis of thousands of cell lines in parallel on a single chip. Here, we describe the ability to perform key cell line development work on the Beacon platform. We demonstrate that commercial production Chinese hamster ovary cell lines can be isolated, cultured, screened, and exported at high efficiency. We compare this process head to head with a FACS-enabled microtiter plate-based workflow and demonstrate generation of comparable clonal cell lines with reduced resources.

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Section: Introductionmentioning

confidence: 99%

A novel mammalian cell line development platform utilizing nanofluidics and optoelectro positioning technology

Tan

Gupta

et al. 2018

Biotechnology Progress

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“…However, in recent years there has been an increased interest in automation and many innovative and low-cost solutions have been proposed and launched. 40 , purification 41 , cloning 42 and transformation/transfection 43,44 . There have been many platforms developed which allow users to conduct these experiments in an integrated manner 45 .…”

Section: Testmentioning

confidence: 99%

An end-to-end solution for automation of biological protocols

Gupta¹

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“…These barriers prevent cross-contamination of arrayed spots and the migration of cells. Some groups have even prepared an integrated microfluidic cell-culture platform that allows for on-chip transfections in small reaction chambers [83,84]. The level of complication and sophistication that has been reached over the years is extensive and has resulted in specialized niches.…”

Section: Reverse Transfectionmentioning

confidence: 99%

“…Only few studies have combined GPCR microarrays with microfluidics so far. In some of those studies, each spot of the cell array was enclosed by a chamber that allowed individual fluidic control [84,103,104]. These micro-chambers mimic a multi-well plate where each well or chamber is individually addressable for measurements by different compounds/ samples.…”

Section: Receptomicsmentioning

confidence: 99%

“…Cohen et al [177] have shown that with increasing DNA levels, the maximum gene expression depends on cell type, the vector used and the method of transfection, so that no single protocol can be applied in all situations. Only few studies have combined reverse transfection with microfluidics [84,[181][182][183]. In those studies, each spot of the cell array was enclosed by a chamber that allowed individual fluidic control.…”

Section: Introductionmentioning

confidence: 99%

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Receptomics, design of a microfluidic receptor screening technology

Roelse¹

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Chapter 1 Introduction Chapter 2 A generic microfluidic biosensor of GPCR-activation-monitoring cytoplasmic Ca 2+ flux in human HEK293 cells Chapter 3 Metabolomics meets functional assays: coupling LC-MS and microfluidic cell-based receptor-ligand analyses Chapter 4 Calcium imaging of GPCR activation using arrays of reverse transfected HEK293 cells in a microfluidic system Chapter 5 The effect of calcium buffering and calcium sensor type on the sensitivity of an array-based bitter receptor screening assay Chapter 6 Statistical models discriminating between complex samples measured with microfluidic receptor-cell arrays Chapter 7 General discussion References Summary Nederlandse samenvatting Abbreviations Acknow ledgements About the author List of publications Overview of completed training activities C H A P T E R 18 Chapter 1 | Introduction Figure 3, Receptor cell assay platforms (diagram adapted from Martins et al 2012). A, The multiwell plate format; cells are seeded on the surface of a multi well plate and (top-down) transfected with receptor coding plasmid DNA. Calcium dye loading, subsequent washing steps and test sample dosing are done using fluid dispensers. The test conditions are multiplexed by using multiple wells and multiple plates making this platform compatible with automation but at relatively high costs and low assay flexibility [90]. B, Receptor cell arrays are created within one well chamber by printing cells [97] or by printing receptor coding plasmid DNA into a micro well and by reverse transfection with cells [81, 85]. Arrays are co-transfected with fluorescent indicator proteins to locate the array. Array washing and sample dosing are done using fluid dispensers. This combines the HTS nature of multi-well plates with the possibility to screen multiple receptors and controls in a single well. C, In microfluidic platforms the cells are cultured or assembled into a microfluidic system [98, 99]. A system can be designed so that different channels address different chambers with (reverse transfected) cells analogous to A but with the option of repeated exposures [84] or a reverse transfected array can be enclosed in one large flowcell, sequentially addressing all spots at once with different samples analogous to B but with the option of repeated challenges and larger arrays [100]. Fluid dispenser Fluid dispenser Adherent cells Substrate Cover layer / flowcell with gasket Fluid Multiwell plate Adherent cell layer Adherent cells Substrate / well plate (A) Well plate HTS (B) Cell microarrays (C) Cell microfluidics Live cell assay platform Multiplexing Single flowcell cell array: Receptomics Multiple chambers Cell array in a well or on a slide Multiple well plates

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A High-Throughput Microfluidic Platform for Mammalian Cell Transfection and Culturing

Cited by 41 publications

References 50 publications

A novel mammalian cell line development platform utilizing nanofluidics and optoelectro positioning technology

A novel mammalian cell line development platform utilizing nanofluidics and optoelectro positioning technology

An end-to-end solution for automation of biological protocols

Receptomics, design of a microfluidic receptor screening technology

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