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Avian - 2002
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| Project Contact: | Douglas Foster | Funding: | $54,650 |
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District:
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| • The Problem • Background • Objectives • Mid-Year Progress Report • Final Report • |
This project needs to build on previous work which isolated cells that could be useful for vaccine production, but which now need to be transformed, using cellular and molecular reagents that have been used successfully in other instances, into cell lines that can continue to grow and divide indefinitely. These continuously growing turkey cell substrates will be used for producing suitably high APV titers for vaccine production.
Presently, there are no certifiable continuously growing (immortal) turkey cell substrates sufficient for the propagation of APV for vaccine production.
Value to the Industry:
Immortalized cell lines can be used in vaccine manufacture, and the cell lines being developed are on target for large scale vaccine production. The immortalized certifiable lines will become even more important to the industry as more regulations involving the generation of vaccines become a reality.
Update – February 2002
Success has been achieved in extending the life spans of turkey kidney and turbinate epithelial cell substrates for analysis of avian pneumovirus (APV) production, but work is still needed to sustain the concentrations of APV virus in those cells needed to make them effective for large scale vaccine production.
APV strain MN2a was propagated in turkey primary kidney or turbinate early passage cells as well as in life span extended cells, and then evaluated in monkey Vero cells. The vaccine concentrations declined as the passage number of the primary cells increased.
Although one clone of turkey turbinate cells was life span extended to passage 36, it produced lowered virus concentrations. Using an earlier passage (<13) of turkey turbinate cells, 80 ml of APV was produced at a nuch higher concentration for in vivo studies.
APV appears to be trapped inside cells by all cell substrates tested. A non-freeze/thaw method of cell lysis (the destruction of a cell by rupture of its membrane) developed here has been shown to dramatically increase the concentration of APV grown in any avian primary or life span extended turkey cell substrates. Using early passage primary turkey kidney cells and this new cell lysis method, this project has produced 60 ml of APV at a substantially increased concentration for in vivo studies.
October 2002
Development of effective vaccines requires the availability of large amounts of a virus (titers) to use in research. Propagating this virus in the laboratory has been difficult, but this work has demonstrated that life span extended turkey epithelial cells can produce amounts of Avian Pneumovirus equal to or greater than the best currently available non-avian alternatives (in cells derived from monkeys). Using a newly developed method to extract the infectious particles from cells, it has also proved possible to significantly increase the quantity of virus derived from several different avian cell substrates. There is continuing progress toward the development of an immortalized turkey epithelial cell substrate for continuous propagation of the virus.