Electrofusion of single cells in picoliter droplets

Rogier M. Schoeman (Corresponding Author), Wesley T.E. Van Den Beld, Evelien W.M. Kemna, Floor Wolbers, Jan C.T. Eijkel, Albert Van Den Berg

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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.

Original languageEnglish
Article number3714
JournalScientific reports
Volume8
DOIs
Publication statusPublished - 27 Feb 2018

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Fusion reactions
Microfluidics
Assays
Microchannels
Platinum
Cells
Throughput
Membranes
Glass
Electrodes
Electric potential
Substrates

Cite this

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title = "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 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.",
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Electrofusion of single cells in picoliter droplets. / Schoeman, Rogier M. (Corresponding Author); Van Den Beld, Wesley T.E.; Kemna, Evelien W.M.; Wolbers, Floor; Eijkel, Jan C.T.; Van Den Berg, Albert.

In: Scientific reports, Vol. 8, 3714, 27.02.2018.

Research output: Contribution to journalArticleAcademicpeer-review

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AU - Van Den Beld, Wesley T.E.

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AU - Eijkel, Jan C.T.

AU - Van Den Berg, Albert

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