brainscattered

thank you for your reply. the accient has been recorded and i have made some inquiries into the nature of the 3rd generation lentiviruses. of course, the accident was stupid and should never happen again, but the probability of serious health hazards seems to be (at least so i can tell in my case) very low. presence of any viral particles after washing and trypsinizing the cells twice already decreases viral titers greatly. the half-life of lentiviruses in cell culture (on acceptor cells, not the producer cell line) is about a day or so and i cultured for several days after removing the viral supernatant. however, half life seems to be increased in cell cultures harbouring macrophages or dedritic cells (internalization of viral particles and subsequent release) moreover, recombination events which could give rise to virus competent of reproduction have not yet been observed (at least 4 recombinations in non-homologous regions would have to take place ...). last but not least, essential genes responsible for pathogenicity of HIV have been deleted - thus, it is impossible to generate wild type HIV just by uncontrolled recombination of the lentiviral vector genome. therefore, the greatest risk seems to be insertional mutagenesis and incorporation of the transgene into the researchers genome. Nonetheless, lifting coverslides with pincers alone is a torture - the coverslides appear to 'stick' to the bottom of the cell culture dish and you have to get underneath somehow (that is what i use the thin, sharp needle for...) - does anyone know a technique (simple as it may be) in order to lift coverslides without help of sharp object such as needles? thank you and cheers

hi, i recently had a stupid accident and am not quite sure what to do now or if the accident could have any aftermath at all? i was handling a primary cell line culture which has previously been transduced with a lentivirus (derived from HIV - 3 vector system (envelope from vsv) + the vector carrying the desired construct). the construct is not oncogenic in nature. after the cells were infected, i changed medium to normal virus-free DMEM and a day later took off the fresh medium, washed the cells with pbs and split them to a 10 cm plate. 2 days later, cells were washed again ad split to 2 new plates containing coverslips. immediately before i took out the coverslips i washed the cells with pbs and thereafter lifted the coverslips using a needle. unfortunately, i pricked myself by accident with that very same needle after scraping around in the cell culture dish in order to lift the coverslips. now i am afraid that i might have introduced virus or virus containing cells into my bloodstream. the cells should be eliminated rapidy and the virus itself should not be capapble of replication, though. is it possible that after so many washing steps there might still have been virus present and if so do i need to worry about infection? after all, the primary cells i was working with should not be able to produce new virus... thank you for your answers.