From: S. Sullivan (sjs93@cam.ac.uk)
Date: Thu Nov 20 2003 - 05:53:44 EST
Nick, Howard et al., I am fusing murine ES cells with somatic cells....on average 10% of the cells fuse. Using a selection regieme, I know at least 1 cell hybrid is formed per 1000 heterokaryons. Nick's question regarding the formation of cell hybrids from heterokaryons....the heterokaryons need to undergo a mitotic event to form hybrids...DNA replicates, the nuclear membrane is broken down and the chromosomes divided and two tetraplod hybrid cells are formed from the heterokaryon. The cell hybrids are genotypically stable (tetraploid) over many passages. I am not concerned with the nuclear protein make up becoming homogenised over time as I will be sorting very shortly after fusion. So when I am looking for hybrid cells, I will have an excess of heterokaryons....using nuclear specific dyes and confocal microscopy would take me forever. Please note that the system does not have to sort hybrids from heterokaryons with 100% efficiency. Even if I could reduce the heterokaryon:cell hybrid ratio by a factor of 10 this would be very useful. Stephen
This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:44:04 EST
