40 Cell Fusion
377
1. Treat mixed cultures of two different ts mutant cells with PEG 6000 Fusion
solution for 1 min at room temperature.
2. Wash the cells three times with serum-free culture medium.
3. Incubate the cells at 24°C for 1 day.
1. Collect the cells, and seed them in heat-treated conditioned medium
in 60 X IS-mm glass dishes at various cell densities.
2. Incubate the cells at 30°C for 12 days.
3. Count the number of colonies.
Results
Using two ts mutants obtained with the procedures outlined above as
parent cell lines, Nakajima and Miyake (1978) made hybrids by means of
the PEG method. Treated cells were selected by a temperature of 30°C,
and the growing cells obtained were hybrids of the two ts mutants.
References
Selection of
hybrid cells
Chiplonkar, J.M. (1992) Electrofusion of mosquito cells. In Vitro Cell. Dev. BioI. 28A, 230-232.
Greene, A.E., Charney, J. Nickols, W.W. and Coriell, 1. (1972) Species identity of insect cell lines. In
Vitro 7, 313-322.
Hsu, H.T. (1978) Cell fusion induced by a plant virus. Virology 84, 9-18.
Miltenburger, H.G., Naser, W.L and Schliermann (1985) Establishment of a lepidopteran hybird cell
line by use of a biochemical blocking method. In Vitro Cell. Dev. BioI. 21, 433-438.
Mitsuhashi, J. (1987) Fusion of insect cells. In: Maramorosch, K. (ed.) Biotechnology in Invertebrate
Pathology and Cell Culture. Academic, New York, pp 387-400.
Morikawa, H., Mitsuhashi, J. and Yamada, Y. (1988) Interkingdom electrofusion between tobacco
mesophyll protoplasts and cultured butterfly cells. Plant Tissue Culture Lett. 5, 90-92.
Nakajima, S. and Miyake, T. (1978) Cell fusion between temperature-sensitive mutants of a Drosophila melanogaster cell line. Somatic Cell Genet. 4,131-141.
Wyss, C. (1979) TAM selection of Drosophila somatic cell hybrids. Somatic Cell Genet. 5,29-37.
Zepp, H.D., Conover, J.H., Hirschhorn, K. and Hodes, H.L. (1971) Human-mosquito somatic cell
hybrids induced by ultraviolet-inactivated Sendai virus. Nat. New BioI. 229,119-121.
377
1. Treat mixed cultures of two different ts mutant cells with PEG 6000 Fusion
solution for 1 min at room temperature.
2. Wash the cells three times with serum-free culture medium.
3. Incubate the cells at 24°C for 1 day.
1. Collect the cells, and seed them in heat-treated conditioned medium
in 60 X IS-mm glass dishes at various cell densities.
2. Incubate the cells at 30°C for 12 days.
3. Count the number of colonies.
Results
Using two ts mutants obtained with the procedures outlined above as
parent cell lines, Nakajima and Miyake (1978) made hybrids by means of
the PEG method. Treated cells were selected by a temperature of 30°C,
and the growing cells obtained were hybrids of the two ts mutants.
References
Selection of
hybrid cells
Chiplonkar, J.M. (1992) Electrofusion of mosquito cells. In Vitro Cell. Dev. BioI. 28A, 230-232.
Greene, A.E., Charney, J. Nickols, W.W. and Coriell, 1. (1972) Species identity of insect cell lines. In
Vitro 7, 313-322.
Hsu, H.T. (1978) Cell fusion induced by a plant virus. Virology 84, 9-18.
Miltenburger, H.G., Naser, W.L and Schliermann (1985) Establishment of a lepidopteran hybird cell
line by use of a biochemical blocking method. In Vitro Cell. Dev. BioI. 21, 433-438.
Mitsuhashi, J. (1987) Fusion of insect cells. In: Maramorosch, K. (ed.) Biotechnology in Invertebrate
Pathology and Cell Culture. Academic, New York, pp 387-400.
Morikawa, H., Mitsuhashi, J. and Yamada, Y. (1988) Interkingdom electrofusion between tobacco
mesophyll protoplasts and cultured butterfly cells. Plant Tissue Culture Lett. 5, 90-92.
Nakajima, S. and Miyake, T. (1978) Cell fusion between temperature-sensitive mutants of a Drosophila melanogaster cell line. Somatic Cell Genet. 4,131-141.
Wyss, C. (1979) TAM selection of Drosophila somatic cell hybrids. Somatic Cell Genet. 5,29-37.
Zepp, H.D., Conover, J.H., Hirschhorn, K. and Hodes, H.L. (1971) Human-mosquito somatic cell
hybrids induced by ultraviolet-inactivated Sendai virus. Nat. New BioI. 229,119-121.