Somatic hybridization of two selected single cells

Koop, Hans‐Ulrich; Dirk, J.; Wolff, D.; Schweiger, Hans−Georg

doi:10.1016/0309-1651(83)90019-x

Cited by 42 publications

(4 citation statements)

References 14 publications

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“…The controlled electrical fusion of selected single gametes was performed using the method described by Koop et al (1983), Koop and Schweiger (1985b), and Schweiger et al (1987). Ten fusion droplets of 1000 nl each, each consisting of 0.55 M mannitol, were overlayered by mineral oil on a coverslip, and selected single gametes were transferred to these droplets for fusion (Fig.…”

Section: Electro Fusionmentioning

confidence: 99%

In vitro fertilization of single, isolated gametes of maize mediated by electrofusion

Kranz¹,

Bautor²,

Lörz³

1991

Sexual Plant Reprod

156

182

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Electrofusion-mediated in vitro fertilization of maize using single sperm and egg cells was performed. Sperm cells were released from pollen grains after rupture of the latter by osmotic shock in the fusion medium (0.55 M mannitol). Egg cells were isolated by enzyme treatment (pectinase, pectolyase, hemicellulase, and cellulase) followed by mechanical isolation. The conditions generally used for the electrical fusion of protoplasts of somatic cells were also applied to the protoplasts of gametic cells of maize. Electrofusion was performed with single pairs of gametes under microscopic observation. The mean fusion frequency was 79%. Isolated egg cells of maize showed protoplasmic streaming during 22 days of culture, but they did not divide. However, after fusion of the sperm with the egg cells, these fused cells did develop, with a mean division frequency of 83%, and grew to multicellular structures. Egg cells and fusion products were cultivated with a maize feeder-cell system.

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Section: Electro Fusionmentioning

confidence: 99%

In vitro fertilization of single, isolated gametes of maize mediated by electrofusion

Kranz¹,

Bautor²,

Lörz³

1991

Sexual Plant Reprod

156

182

View full text Add to dashboard Cite

show abstract

“…Electrofusion of selected protoplasts was performed as described in detail elsewhere (Koop et al 1983;Koop and Schweiger 1985 b;Schweiger et al 1987;Kranz et al 1991). Fusion was induced by applying a single or two to three negative DC pulses (50 gs; 0.7-1.0 kV cm-1) after dielectrophoretic alignment of defined gametes and cytoplasts (1 MHz, 48-71 V cm-1) on one of the electrodes for a few seconds using an electrofusion apparatus (CFA 400, Krfiss, Hamburg, FRG).…”

Section: Electro Fusionmentioning

confidence: 99%

Electrofusion-mediated transmission of cytoplasmic organelles through the in vitro fertilization process, fusion of sperm cells with synergids and central cells, and cell reconstitution in maize

Kranz¹,

Bautor²,

Lörz³

1991

Sexual Plant Reprod

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Fusion products were created by the electrofusion of single sperm cells with single synergids and central cells. The synergid was also fused with the sperm cell, occasionally in the presence of adhering second synergids, egg cells, and central cells. Single egg cells were fused with single sperm cells in the presence of adhering synergids and the central cell. Cytoplasmic organelles were transmitted through the fertilization process by electrofusion using cytoplasts of maize mesophyll cells. Cell reconstitution was achieved by fusion of one or two sperm cells with single enucleated protoplasts, thus creating a haploid or a diploid cell.

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“…caused by needle electrodes, may circumvent these problems. This approach is feasible only in combination with individual, single cell fusion systems (Koop et al 1983).…”

Section: Microprotoplastsmentioning

confidence: 99%