Electrofusion of single cells in picoliter droplets

Abstract We present a microfluidic chip that enables electrofusion of cells in microdroplets, with exchange of nuclear components. It is shown, to our knowledge for the first time, electrofusion of two HL60 cells, inside a microdroplet. This is the crucial intermediate step for controlled hybridoma...

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Autores principales:	Rogier M. Schoeman, Wesley T. E. van den Beld, Evelien W. M. Kemna, Floor Wolbers, Jan C. T. Eijkel, Albert van den Berg
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3606b79671de4d729848e07db56bd288
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spelling	oai:doaj.org-article:3606b79671de4d729848e07db56bd2882021-12-02T15:08:51ZElectrofusion of single cells in picoliter droplets10.1038/s41598-018-21993-82045-2322https://doaj.org/article/3606b79671de4d729848e07db56bd2882018-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-21993-8https://doaj.org/toc/2045-2322Abstract We present a microfluidic chip that enables electrofusion of cells in microdroplets, with exchange of nuclear components. It is shown, to our knowledge for the first time, electrofusion of two HL60 cells, inside a microdroplet. This is the crucial intermediate step for controlled hybridoma formation where a B cell is electrofused with a myeloma cell. We use a microfluidic device consisting of a microchannel structure in PDMS bonded to a glass substrate through which droplets with two differently stained HL60 cells are transported. An array of six recessed platinum electrode pairs is used for electrofusion. When applying six voltage pulses of 2–3 V, the membrane electrical field is about 1 MV/cm for 1 ms. This results in electrofusion of these cells with a fusion yield of around 5%. The operation with individual cell pairs, the appreciable efficiency and the potential to operate in high-throughput (up to 500 cells sec−1) makes the microdroplet fusion technology a promising platform for cell electrofusion, which has the potential to compete with the conventional methods. Besides, this platform is not restricted to cell fusion but is also applicable to various other cell-based assays such as single cell analysis and differentiation assays.Rogier M. SchoemanWesley T. E. van den BeldEvelien W. M. KemnaFloor WolbersJan C. T. EijkelAlbert van den BergNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-8 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rogier M. Schoeman Wesley T. E. van den Beld Evelien W. M. Kemna Floor Wolbers Jan C. T. Eijkel Albert van den Berg Electrofusion of single cells in picoliter droplets
description	Abstract We present a microfluidic chip that enables electrofusion of cells in microdroplets, with exchange of nuclear components. It is shown, to our knowledge for the first time, electrofusion of two HL60 cells, inside a microdroplet. This is the crucial intermediate step for controlled hybridoma formation where a B cell is electrofused with a myeloma cell. We use a microfluidic device consisting of a microchannel structure in PDMS bonded to a glass substrate through which droplets with two differently stained HL60 cells are transported. An array of six recessed platinum electrode pairs is used for electrofusion. When applying six voltage pulses of 2–3 V, the membrane electrical field is about 1 MV/cm for 1 ms. This results in electrofusion of these cells with a fusion yield of around 5%. The operation with individual cell pairs, the appreciable efficiency and the potential to operate in high-throughput (up to 500 cells sec−1) makes the microdroplet fusion technology a promising platform for cell electrofusion, which has the potential to compete with the conventional methods. Besides, this platform is not restricted to cell fusion but is also applicable to various other cell-based assays such as single cell analysis and differentiation assays.
format	article
author	Rogier M. Schoeman Wesley T. E. van den Beld Evelien W. M. Kemna Floor Wolbers Jan C. T. Eijkel Albert van den Berg
author_facet	Rogier M. Schoeman Wesley T. E. van den Beld Evelien W. M. Kemna Floor Wolbers Jan C. T. Eijkel Albert van den Berg
author_sort	Rogier M. Schoeman
title	Electrofusion of single cells in picoliter droplets
title_short	Electrofusion of single cells in picoliter droplets
title_full	Electrofusion of single cells in picoliter droplets
title_fullStr	Electrofusion of single cells in picoliter droplets
title_full_unstemmed	Electrofusion of single cells in picoliter droplets
title_sort	electrofusion of single cells in picoliter droplets
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/3606b79671de4d729848e07db56bd288
work_keys_str_mv	AT rogiermschoeman electrofusionofsinglecellsinpicoliterdroplets AT wesleytevandenbeld electrofusionofsinglecellsinpicoliterdroplets AT evelienwmkemna electrofusionofsinglecellsinpicoliterdroplets AT floorwolbers electrofusionofsinglecellsinpicoliterdroplets AT jancteijkel electrofusionofsinglecellsinpicoliterdroplets AT albertvandenberg electrofusionofsinglecellsinpicoliterdroplets
_version_	1718387990864068608

Electrofusion of single cells in picoliter droplets

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