Differentiation of Non-adherent Bone Marrow Cells to Mast Cells by Supernatant of Spleen Cells Culture Medium in Rat.

Objective: Mast cells are connective tissue cells that mostly known for their role in allergic and other inflammatory disease. They play important role in immediate allergic reactions and inflammatory responses. There are evidences illustrating the involvement of these cells in other numerous physiological and pathophysiological reactions. The aim of the present study was to establish a simple and affordable method for the differentiation and culture of murine mast cells from rat bone marrow. Materials and Methods: Mast cells were generated from bone marrow of 10-15 weeked aged male mice based on a modified method change of the tissue culture flasks from 75-cm2 to 25-cm2 via supernatant of the spleen cell culture medium. The granularity of the mast cells was determined using toluidine blue staining. Mast cells were characterized and analyzed using Mast Cell Tryptase Antibody and CD 117, CD 34. Tryptase activity was present only in mast cell. It is helpful method to distinguish the activation of mast cell from other cells as neutrophils. Results: Mast cells were generated from bone marrow of male rat. The cells were positive for Mast cell-related antigens for each of CD117 and Tryptase, and negative for CD34. Conclusions: In the present study change of the tissue culture flasks from 75-cm2 to 25-cm2 ended up more dense arrangement of the cells and subsequently the effect of the secreting factors like IL4, IL3 and SCF was enhanced and improved. [ABSTRACT FROM AUTHOR]