Chapter 40
Cell Fusion
Subprotocol 1
Cell Fusion by Viruses
In early cell fusion studies, viruses were mostly used as fusogens. There
are many viruses that produce polynucleated cells. Of the insect viruses,
iridoviruses and densonucleosis virus are known to induce cell fusion.
Hemagglutinating virus of Japan (HVJ) was first used as a fusogen in
mammalian cell culture. This virus can also be used as a fusogen for
cultured insect cells. The following procedure is based on the report of
Zepp et al. (1971) on inter phylum hybrid formation.
Materials
- Centrifuge and centrifuge tubes
- Automatic shaker
- Water bath
- Culture flask
- MEM supplemented with 10% FBS
- Grace's medium (see Appendix 1)
- HeLa cells cultured in Eagle's MEM
- Grace's Ae cells cultured in Grace's medium (cf. Appendix 2)
- HVJ (also called Sendai virus) inactivated by UV light
Materials
PROTOCOL
Equipment
Solutions
Cells
Viruses
l. Culture HeLa cells in a monolayer at 37°C in Eagle's MEM supple- Parent cell
mented with 10% FBS.
culture
2. Culture Ae cells in a suspension culture at 26°-28°C in Grace's medium supplemented with 10% FBS.
l. Harvest approximately 4 x lOs cells from both cultures and suspend Fusion
the cells in their culture media respectively.
2. Mix the suspension of both cells in a centrifuge tube.
3. Centrifuge the cell mixture at 250 g for 5 min.