Development of Techniques about Production of Recombinant vaccines and cells with Activated Immunogenic potential

Abstract

Methods about derivation of myeloid-like and lymphoid-like cells from normal embryonic fibroblasts were developed and tested. Balb/c mice normal mouse embryonic cells from line 3T3 were co-cultivated with mouse malignant myeloma cells, containing inserted. Murine Leukemia Virus (MuLV) (with RNA-genome, Retroviridae family). For this goal, separate subpopulation of normal mouse embryonic fibroblasts were pre-incubated in cultural fluid, supplemented by previous incubation of myeloma cells in it, after subsequent centrifugation and filtration. Other sub-populations of 3T3 cells were co-cultivated with myeloma cells, by addition of cultural fluids plus cellular suspensions of both types. Subsequently, all mixed cultures were freezed after addition of cryo-protector Dimethylsufoxide (DMSO), subsequently thawed and re-incubated in standard laboratory conditions. In the cultures, pre-incubated in cultural fluid, supplemented by previous incubation of mouse malignant myeloma cells, cells in different stages of myeloid/phagocyte and lymphoid/plasmatic cell differentiation were observed, but in addition of suspensions of cells from both types, appearance of hybrid cells of myeloid-like and phagocyte-like, as well as of lymphoid-like and plasmatic cells-like with mouse malignant myeloma cells were noted. The established changes could be explained with the eventual existence of capable to differentiate to various lineages stem-like cellular sub-populations in the general 3T3 cell line. Also, activated fusion between different cells, as well as between cells and viral particles on the influence of DMSO and of the drastic temperature changes was proposed, which could also lead to transfer of nucleotide sequences. On this principle, a possibility for production of recombinant viral vaccines by exchange of nucleotide sequences between cells and viral particles could be suggested. Furthermore, in confirmation of the literature findings, a capability of non-myeloid and non-lymphoid cells to produce membrane receptor glycoproteins was proposed.