Microcarriers as a culturing system of insect cells and insect viruses

There is an increasing interest in culturing of insect cells, which are host for arthropod-born (Arbo) viruses. The potential applications of Arbo viruses are in the following two main fields: 1. Medical applications (e.g. preparation of viral vaccines and viral antigens for diagnostic purposes). 2. As bioinsecticides in pest control in horticulture, agriculture and forestry. One of the potential cell substrates for these applications is an anchorage-dependent-mosquito cell line established from embryonic tissues of Aedes aegypti (AA). The following areas were investigated in the reported research with the AA cell line: The AA cells were successfully propagated in microcarrier (MC)-culturing-systems. Of the tested MC's the cellulose-based microgranular MC (DE-53 of Whatman, having an exchange capacity of 2 meq/g) was found to be the best MC. Cells grew in MCs-cells aggregates in submerged spinner culture. The AA-MC's culture was successfully scaled-up to 8 litre culture volume. "Trypsinization" of the AA cells from the MC surface is successfully done by RDB, a dispersion agent from a plant origin (produced and marketed at the author's Institute). Other known dispersion agents (trypsin, collagenase, pronase) failed to disperse the AA cells from the MC. A serum-free medium was developed for culturing the AA cells on the DE-53 MC's. Bovine serum albumin was the main serum substituant in the developed medium. Arboviruses, from the Toga group, were grown in the AA-MC culture. Sindbis virus (from alpha-group) and West Nile virus (from the Flavi group), chronically infected the AA cells, which continuously produce and liberate these two viruses.(ABSTRACT TRUNCATED AT 250 WORDS)