Siroký J, Nebola M, Pribyla L, Karpfel Z
Institute of Biophysics, Czechoslovak Academy of Sciences, Brno.
Gen Physiol Biophys. 1987 Oct;6(5):439-48.
A new apparatus was constructed which enables the use of the electrofusion method to obtain polynuclear cells of various mammalian cell lines, erythrocytes and plant protoplasts. This technique was applied to both suspensions and monolayers. Electrical and other physical parameters were monitored to find optimal conditions for mutual contact of cells (dielectrophoresis) and subsequent fusion. In the suspension technique, dielectrophoresis of mouse erythrocytes occurred at a field frequency of 20 kHz and a strength of 500 V.cm-1, whereas cultured mammalian cells and plant protoplasts required a frequency of 1-1.4 MHz and a strength of 250-800 V.cm-1. Fusion of cells was induced after the application of 1 to 10 high-voltage pulses of 1-5 kV.cm-1, 10-36 microseconds duration. After these high-voltage pulses were to the monolayer of mouse L cells, about 12% viable homokaryons were obtained.
构建了一种新装置,该装置能够利用电融合方法获得各种哺乳动物细胞系、红细胞和植物原生质体的多核细胞。此技术应用于悬浮液和单层细胞。监测电和其他物理参数以找到细胞相互接触(介电电泳)及随后融合的最佳条件。在悬浮技术中,小鼠红细胞的介电电泳发生在20 kHz的场频和500 V.cm-1的强度下,而培养的哺乳动物细胞和植物原生质体需要1 - 1.4 MHz的频率和250 - 800 V.cm-1的强度。在施加1至10个1 - 5 kV.cm-1、持续时间为10 - 36微秒的高压脉冲后诱导细胞融合。将这些高压脉冲施加到小鼠L细胞单层后,获得了约12%的存活同核体。