A biophysical approach to the optimisation of dendritic-tumour cell electrofusion

Journal Article · Fri Aug 04 00:00:00 EDT 2006 · Biochemical and Biophysical Research Communications

DOI:https://doi.org/10.1016/j.bbrc.2006.05.193· OSTI ID:20854390

Sukhorukov, Vladimir L ^[1]; Reuss, Randolph ^[1]; Endter, Joerg M ^[1]; Fehrmann, Steffen ^[1]; Katsen-Globa, Alisa ^[2]; Gessner, Petra ^[1]; Steinbach, Andrea ^[1]; Mueller, Kilian J ^[1]; Karpas, Abraham ^[3]; Zimmermann, Ulrich ^[1]; Zimmermann, Heiko ^[2]

---

1. Lehrstuhl fuer Biotechnologie, Biozentrum, Universitaet Wuerzburg, Am Hubland, D-97074 Wuerzburg (Germany)
2. Abteilung Kryobiophysik and Kryotechnologie, Fraunhofer-Institut fuer Biomedizinische Technik, 66386 St. Ingbert (Germany)
3. Department of Haematology, Cambridge University, Medical Research Council Centre, Hills Road, Cambridge CB2 2QH (United Kingdom)

Electrofusion of tumour and dendritic cells (DCs) is a promising approach for production of DC-based anti-tumour vaccines. Although human DCs are well characterised immunologically, little is known about their biophysical properties, including dielectric and osmotic parameters, both of which are essential for the development of efficient electrofusion protocols. In the present study, human DCs from the peripheral blood along with a tumour cell line used as a model fusion partner were examined by means of time-resolved cell volumetry and electrorotation. Based on the biophysical cell data, the electrofusion protocol could be rapidly optimised with respect to the sugar composition of the fusion medium, duration of hypotonic treatment, frequency range for stable cell alignment, and field strengths of breakdown pulses triggering membrane fusion. The hypotonic electrofusion consistently gave a tumour-DC hybrid rate of up to 19%, as determined by counting dually labelled fluorescent hybrids in a microscope. This fusion rate is nearly twice as high as that usually reported in the literature for isotonic media. The experimental findings and biophysical approach presented here are generally useful for the development of efficient electrofusion protocols, especially for rare and valuable human cells.

• OSTI ID: 20854390
• Journal Information: Biochemical and Biophysical Research Communications, Vol. 346, Issue 3; Other Information: DOI: 10.1016/j.bbrc.2006.05.193; PII: S0006-291X(06)01257-5; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); ISSN 0006-291X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
ALIGNMENT
ANIMAL CELLS
BIOLOGICAL STRESS
BLOOD
DIELECTRIC MATERIALS
HYBRIDIZATION
MICROSCOPES
OPTIMIZATION
SACCHAROSE
SORBITOL
TIME RESOLUTION
VACCINES