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Electrofusion of B-cells and cancer cells on chip
Electrofusion of B-cells and cancer cells on chip
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Figure 1: schematic representation of electrofusion of a b-cell and a myeloma cell |
The aim of this project is to increase the electrofusion-efficiency of myeloma and B-cells using microtechnology. Currently, there is an interest in the development of antibodies for research as well as diagnostic and therapeutic purposes. One of the techniques to generate antibodies is the electrofusion of antibody producing cells (B-cells) with cancer cells. The fusion product (hybridomas) have ‘the best of both worlds’: the infinite production of a certain antibody (figure 1). Electrofusion occurs under influence of an electric field in a fusion chamber. This technique using a conventional fusion chamber functions well, however the method is very laborious and the final efficiency is relatively low. This accounts for the fusion efficiency (number of hybridomas) as well as the production efficiency of the right antibody by the created clone. The aim is to increase electrofusion efficiency by microtechnogly, via selective pairing of the cells, bringing the membranes in close proximity and having a high throughput system.
There are currently two approaches which are investigated to achieve efficient electrofusion on chip.
Cell pairing trap array
A cell pairing a trap array (800 traps) was designed based on an article by Skelley et al 2009 (figure 2). After loading of both cell types, first the B-cells, second the myeloma cells, pairs are formed. Subsequently, one or more pulses are applied and fusion efficiency will be determined.
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Figure 2: overview of trap array chip on the left (SEM image). Right shows top view of trapped B-cells (Hoechst staining, blue) and myeloma cells (cytoplasmic stain, green). Black structures are electrodes |
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Electrofusion in a droplet
Second, our focus is also set on electrofusion in a droplet (figure 3A), which highlights the high throughput character. Furthermore, the droplet platform can be easily integrated in respect to screening (analysis) and selection (sorting) of the formed hybridomas. Cells are encapsulated in an isolated volume and when de droplet is small enough, cells are in good alignment to electrofuse. The first design is a PDMS-glass chip for the generation of droplets using hydrodynamic focusing. The short term goal is electrofusion in droplets, focusing on characterizing of droplet generation, embedding beads and cells in droplets and sorting the cell containing droplets to increase fusion efficiency (see figure 3C).
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Figure 3. A schematic overview of electrofusion in a droplet. B Table of different ratios (the water flow rate was fixed at .25 μl/min). Corresponding microscopic images of droplet generation on chip (scale bar is 100 μm). C. Beads in a droplet. Corresponding to Poisson statistics (lambda =0.25, 19% of droplets containing 1 cell). |
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INTERESTED?
Are you interested and would you like to get in contact with the multidisciplinary environment of the BIOS group, please contact me through the following e-mail address.
Skelley, A. M., O. Kirak, et al. Nature Methods Microfluidic control of cell pairing and fusion. 2009
CONTACT INFORMATION
Evelien Kemna
MESA+ Institute for Nanotechnology
University of Twente
7500 AE Enschede
The Netherlands
Phone: +31 (0)53 489 2755
Email: e.w.m.kemna@ewi.utwente.nl
