Preparation of honey bee (Apis mellifera L.) primary cell culture by enzymatic disaggregation and planting minced tissues methods

In this study, in order to preparation of primary cell culture of honey bees, some samples were collected from larval and pupal honey bee and were prepared according to standard guidelines. For preparing of primary culture, the short-time enzymatic disaggregation (three-phase, 10 min), long-time (18 h) and planting minced tissues methods were used. For growth of obtained cells in various methods, the Grace medium enriched with fetal bovine serum (FBS) to a value of 15% was used in the temperature 28oC. Cells obtained in various methods, had good status in number and rate of growth of living cells and have reached in full confluency after 2-5 weeks and several successive subcultures were prepared. In enzymatic disaggregation method the best result was obtained from long-time trypsinization of pupa stage. The result of planting minced tissues method in the larval and pupal stages were similar and both were great.