Our new device delivers electrifying results
Spontaneous fusion of somatic cells in a test tube occurs less than 1 in 1,000,000 times. Though this is good if you are a living organism, it is sometimes desirable to fuse cells for specialized applications. Historically, a chemical such as PEG has been used to increase spontaneous fusion by 2 or 3 orders of magnitude; alternatively, inactivated viruses have been used to promote fusion of proximal cells to an equivalent level. More recently, protocols utilizing AC and DC electric fields have further improved the efficiency of cell fusion – in some cases, by as much as 25 to 100-fold.
The ECFG21 incorporates the latest in solid state electronic design for a robust system with plenty of useful feedback. The two phase technique relies upon an AC field to physically align the cells end-to-end in a chain formation. Once in they’re in physical contact, the system applies a series of DC pulses to disrupt the plasma membrane and induce fusion of adjacent cells. Over the next 30 minutes as many as one in every hundred cells makes a viable hybrid cell with two nuclei. The nuclei later fuse and the process completes. Only the ECFG21 can provide real-time feedback of voltage and current to ensure that every sample is under optimal conditions for cell fusion. The ECFG21 precisely controls AC to DC switching time, the DC pulsing amplitude and length (including a convenient reverse polarity option), post fusion AC current for maintenance of the end-to-end cell configuration. It even offers manual control of DC pulsing — useful when observing first time cell optimizations under a microscope.
Reviews of this new system have been tremendous. It’s been reported that the ECFG21 provides much better control of experimental conditions as compared with chemical and viral methods. This is due to the electric field feedback; the system can determine in real time if ionic conditions are not optimal for fusion, as high ion concentrations create a physical streaming of solution and disruption of cell chains. Electro-cell fusion typically produces hybrid cell lines that are more stable, in part because of the lower cytotoxicity inherent in the precise electric field and pulsing control of this instrument. Results tend to be more reproducible and repeatable, too. And don’t forget the human element, either! Here at Bulldog, we will work closely with you to optimize your results, and we have years of experience doing so. Give us a ring and start fusing immediately!
New vistas for research
Somatic cell fusion is a tricky procedure, traditionally used for making hybridomas for monoclonal antibodies. This classic technique fuses an immortal myeloma cell line with a spleen-derived antibody-producing B-cell. This hybridoma cell line will stably produce a specific monoclonal anitbody for months or years. Cell fusion has also been used to ask fundamental questions regarding gene expression and to study cancer progression. For example, ES/EG cells have been fused to T-cells to study T-cell activation by reprogramming pluripotent stem cells. This application also enables the amplification of viral particles in non-host cells, or can be used to study how a cell divides, and has been used to great success to fuse oocytes and somatic cells. There are many other lesser-known applications, such as fusing yeast and mold cells, plant protoplast electro-cell fusion, etc. The potential for you to create your own unique hybrid cell line to share with the rest of the research community is nearly limitless.
Hybridomas show their love for the ECFG21
One of our customers, Precision Antibody, has spent years developing highly optimized cell fusion protocols based upon the classic PEG method. Their procedures have resulted in the production of first-rate antibodies for their customers. When they asked us to use the ECFG21 to tackle their most difficult antigens, we were cautiously optimistic. In fact, when the final results were tallied, everyone was floored by the dramatic improvements provided by the electro-fusion technology. They tested three different categories of antigens, and the ability of the ECFG21 to improve the number of hybridomas produced therein. In addition, they calculated how many of these were in fact positive for reactivity with the antigen. The first, a rather large 20kDa immunosuppressive protein, gave fair results with PEG for making hybridomas, but could not produce even a single positive clone that was reactive with the antigen. The second and third antigens tested were both short (8 and 17 amino acids respectively) and hydrophobic. Again they produced a low number of hybridomas, but very few positive reactive clones when using the PEG method. The resulting number of hybridomas using the ECFG21 was 25x to 100x higher. More impressively, the number of hybridoma clones that were positive with the antigen (which is the most important consideration), was a staggering 50x to over 200X higher. If your lab is looking to consistently produce specific monoclonal antibodies, the ECFG21 isn’t just good—it’s your only sensible choice.
Somatic-Cell Nuclear Transfer (SCNT)
The ECFG21 is more than capable of reprogamming somatic cells using enucleated oocytes for animal cloning.
Tetraploid embryo complementation
See the ECFG21 used for tetraploid chimera production using a two-step cloning procedure to produce mice from cancer cells.
These three antigens were all classified as “difficult to obtain positive clones”. The ECFG21 vastly increased the number of clones in all cases, and in at least one case, resulted in positive clones when none resulted with the traditional PEG method.
1) Antigen used: 20 kDa immunosuppressive protein with high homology
2) Antigen used: Hydrophobic octapeptide
3) Antigen used: Highly hydrophobic 17 amino acid peptide.
|Fusion/M:||Number of hybridomas generated per 1 x 106 lymphocytes|
|Positive clone/M:||Number of clones producing monoclonal antibodies against the antigen per 1 x 106 lymphocytes|
|E w/P2:||ECFG21 electrofusion performed using CUY497P2 electrode (up to 0.8 mL)|
|E w/P8X10:||ECFG21 electrofusion performed using CUY497P8X10 electrode (up to 6.4 mL)|
Data provided courtesy of Jun Hayashi, Ph.D., VP of Precision Antibody
C115CB-2 – 2mm Female to 4mm Male plug & Cover banana plug cable, red & black, 2 meter$149.00 Add to cart
Electrodes and Accessories for Cell Fusion
CUY497P2 – ECFG21 Bath w/platinum plate electrodes on slide glass, 2mm gap, L80 x W2 x H5mm, 0.8ml for cell fusionRequest Quote
CUY497P8 – ECFG21 Bath w/platinum plate electrodes on slide glass, 8mm gap, L80 x W8 x H5mm, 3.2ml for cell fusionRequest Quote
CUY497P8X10 – ECFG21 Bath w/platinum plate electrodes on slide glass, 8mm gap, L80 x W8 x H10mm, 6.4ml for cell fusionRequest Quote
CUY497P10X10 – ECFG21 Bath w/platinum plate electrodes on slide glass, L80 x W10 x H10mm, 8.0ml for cell fusionRequest Quote
Electrodes and Accessories for Activation and Nuclear Transfer
CUY5000P05 – Platinum Wire Petridish Electrode for Cloning Activation, 0.5mm gap, compatible, compatible w/ ECFG21 & NEPA21Request Quote
CUY5000P1 – Platinum Wire Petridish Electrode for Cloning Activation, 1mm gap, compatible w/ ECFG21 & NEPA21Request Quote
CUY5100-100 – Z-Shape Cloning Electrode for Somatic Cells & Fertilized Eggs, 5mm Length x 100µm Diameter Tip, 2/pkg, Compatible w/ECFG21 & NEPA21Request Quote
|Voltage||0 – 80 Vrms|
|Post-Fusion Duration||0-99 seconds|
|Post-Fusion Decay Mode||On/Off|
|Pause between AC / DC||5 µsec|
|Pulse Wave||Square Wave|
|Voltage||1 – 1500 V|
|Pulse Length||1-99 µsec|
|Pulse Interval||0.1 – 9.9 sec|
|Number of Pulses||0 – 99|
|Decay Rate||0 – 99%|
|Impedance Measurement||0.01 – 50 kΩ|
|Operation Mode||Automatic / Manual|
|Power Requirements||AC100-240V 50/60Hz|
|Size||386(W) x 370(D) x121(H) mm|