TECHNICAL INFO GenomONE-CF EX

For Laboratory Research Use Only. Not for Use in Clinical Diagnosis or Therapeutic Procedures.

	Product Name	Cat #	Size	Price
	GenomONE-CF EX	CF001EX	1kit (HVJ-E 1vial set)	distributor website

GenomONE-CF EX is a cell fusion kit composed of HVJ Envelope (HVJ-E) and special buffers.

It can be used with both adhering cells and floating cells. Fusion of cells of the same
or different types is possible with this kit in only 30 minutes.

Cell fusion with GenomONE-CF EX (HVJ-E Cell Fusion Kit) (time course of 67 minutes) BHK-21 cells labeled with red and
　　 green fluorescent dyes incubated at 37°C in the presence of HVJ-E and observed under a confocal laser scanning
　　 microscope

What's HVJ Envelope (HVJ-E)?

[Features]
This kit resolves the following problems, encountered when attempting cell fusion using
polyethylene glycol (PEG) or electrofusion

Operation is too complex

　Toxicity is high and the efficiency of cell fusion low

　Reproducibility is low

　Much time is needed to optimize fusion conditions

　Rapid or simultaneous processing of many samples is not possible due to the necessity
for strict control of reactions

[Principle]
Principle for cell fusion triggered by HVJ-E

A local model of two cells cross-linked by one HVJ-E particle.
Step 1	If HVJ-E is added in amounts of more than several hundred HVJ-E per cell at low temperatures (0-8°C), HVJ-E is immediately adsorbed on the cell surface mediated by the receptor (acetyl type sialic acid recognized by HN protein) (Step 1), and cells undergo agglutination cross-linked by HVJ-E particles (Step 2). At this stage, the hydrophobic domain at the N-terminal of cleaved F protein (F1) penetrates into the double lipid layer of the cell membrane, causing distortion of the membrane severe enough to allow an inflow of ions.
Step 2
Step 3	If this cell / HVJ-E complex is heated at 37°C, the distortion of the cell membrane is further expanded, accompanied by temporary alteration of the cell membrane lining structure. This change is transient and the membrane soon returns to its normal structure. However, if a strong hydrophobic connective force is applied at this stage, fusion between cell membranes takes place (Step 3).

[Reference]
Okada Y., et al., Exp. Cell Res., 93, 368-378 (1975).
Okada, Y., Cell Fusion and Life Science, Brain Center (2001).

[Applications]

No.	Reference	Fused cell	Application
1	Journal of Immunology, 173, 4297-4307 (2004).	Dendritic ｃells and B16BL6 melanoma cells	Cancer immunology
2	Mutation Research, 562, 11-17 (2004).	Nornal human fibroblasts (G0/G1) and HeLa (M phase)	Premature chromosome condensation (PCC)
3	Radiation Research, 165, 59-67 (2006).	Nornal human fibroblasts (G0/G1) and HeLa (M phase)	Premature chromosome condensation (PCC)
4	*Chromosome Science*, 9, 65-73 (2006).	Microcell and HT1080 Microcell and hiMSC (immortalized human mesencymal stem cell)	Microcell mediated chromosome transfer
5	*Hybridoma (Larchmt)*, 26, 381-385 (2007).	Rat lymph node cells (immunized) and mouse myeloma cells (P3U1)	Preparation of monoclonal [Immunogen: recombinant TARSH-CH protein]
6	*Immunol. Lett.,* 121, 97-104 (2008).	Rat lymph node cells (immunized) and mouse myeloma cells (P3U1)	Preparation of monoclonal antibodies
7	Human Reproduction, 23, 1377-1384 (2008).	Mouse enucleated oocyte and nucleus	Nuclear transfer
8	*Biosystems,* 92, 226-232 (2008).	Rat cardiac myocute and Rat cardiac fibroblast	Analysis of cellular function
9	*Journal of Cell Science*,122, 912- 918 (2009).	HeLa	Analysis of cellular function
10	*Int. J. Oncol*., 35, 249-255 (2009).	Dendritic ｃells and Meth A cells (BALB/c-derived fibrosarcoma)	Cancer immunology
11	*Nature,* 461, 367-372 (2009).	Macaca mulatta enucleated oocyte and nucleus	Mitochondrial gene replacement
12	Reproduction, 137(4), 625-632 (2009).	Mouse enucleated oocyte and nucleus	Nuclear transfer
13	*Biochem. Biophys. Res. Commun.,* 390, 547-551 (2009).	Mouse spleen cells (immunized) and mouse myeloma cells (P3U1)	Preparation of monoclonal antibodies [Immunogen: synthetic human IDH1R132H peptide]
14	Biochem. Biophys. Res. Commun., 391, 750-755 (2010).	Mouse spleen cells (immunized) and mouse myeloma cells (P3U1)	Preparation of monoclonal antibodies [Immunogen: lacto-series gangliosides 30-isoLM1 and 30,60-isoLD1]
15	Stem Cells Dev., 19(2), 229-238 (2010).	Mouse adult bone marrow mononuclear cells and Mouse Es cells Mouse embryonic fibroblasts (from Oct4-GFP transgenic mice) and Mouse ES cells	Reprogramming of somatic cells
16	*Biochem. Biophys. Res. Commun*., 394, 1053-1057 (2010).	Mouse embryonic fibroblasts and Mouse ES cells	Reprogramming of somatic cells
17	*Nature*, 465, 82–85 (2010).	Human zygotes	Pronuclear transfer
18	*Nature Protocols*, 5(6), 1138-1147 (2010).	Macaca mulatta enucleated oocyte and nucleus	Mitochondrial gene replacement
19	*Hybridoma (Larchmt)*, 29(1), 7-11 (2010).	Rat lymph node cells (immunized) and mouse myeloma cells (P3U1)	Preparation of monoclonal antibodies
20	*Stem Cells.* 28(8), 1338-1348 (2010).	Human Es cells and Human fibroblasts	Reprogramming of somatic cells
21	*Vet. Immunol. Immunopathol.,* 138(3), 231-234 (2010).	Mouse lymphoid cells (immunized) and mouse myeloma cells(P3X63Ag8.653)	Preparation of monoclonal antibodies [Immunogen: recombinant feline cystatin C]
22	*Radiat. Res.,*174(5), 566-573 (2010).	Human fibroblasts	Premature chromosome condensation (PCC)
23	*FASEB J.,* 25(3), 830-839 (2011).	Human adipose tissue-derived stem cells and Neonatal rat cardiomyocytes	Analysis of cellular function
24	*Biochem. Biophys. Res. Commun.,* 406(4), 608-613 (2011).	Mouse spleen cells (immunized) and mouse myeloma cells (P3U1)	Preparation of monoclonal antibodies
25	*Biol. Reprod.,* 85, 70-77 (2011).	Mouse enucleated oocyte and nucleolus-like body (NLB)	GV transplantation

[Specification]

Cat. #

Freeze-dried HVJ-E*¹(inactivated HVJ)

0.26mL/vial

(when reconstituted

HVJ-E suspending buffer

0.5mL/vial

Cell fusion buffer

(20 X concentrate)*²

10mL/vial

CF001EX

1 vial

*¹HVJ-E: HVJ Envelope

Moisture can reduce activity. Store the product in a refrigerator, sealed in an aluminum package.

*² Before use, dilute 1:20 with water (e.g.: endotoxin-free water for injection)

[Frequency of use]
For fusion of cells of the same or different types

CF001EX (HVJ-E:1 vial) : 100 runs

For preparing B cell hybridoma

CF001EX (HVJ-E:1 vial) : 10 runs

[Stability during storage]

[Quality assured period of freeze-dried HVJ-E] : See the aluminum package for HVJ-E.
[HVJ-E suspension prepared with HVJ-E suspending buffer]:

             For continuous use, store in a refrigerator (2-8°C) for up to two weeks.
             For extended storage, freeze in working aliquots at -80°C for up to 3 months.
             Thawing after freezing is possible only once. HVJ-E suspending buffer and Cell fusion Buffer should be stored
             in a refrigerator (2-8°C).

[Related Technical Information]

Instruction manual	Safety Information
Data sheet	Application (Nuclear Transplantation)
Application (Hybridoma production)	FAQ
Trouble shooting	Reference list
Movie (Experimental protocol for cell fusion)