Developing Rat Bone Marrow Derived Mast Cells by the Splenic Cells Culture Supernatant of Rat and Mouse

Authors

1 Department of Comparative Histology and Anatomy Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran,

2 Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran,

3 Department of Immunology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Abstract

Background: Mast cells play a critical role in the pathogenesis of various immunological and non-immunological diseases. It is now accepted that culturing primary mast cells considered as a tool for investigation role of mast cells in diseases. Development of various animal primary mast cells and their function could be used for the translational studies in the pathogenesis of human diseases. The aim of the study was to develop simple and cost-efficient method for differentiation and culture of rat mast cells from bone marrow by using rat and mouse spleen supernatant. Materials and Methods: Bone marrow cells from 10 to15-weeks-old male rats was obtained and cultured for three weeks on cell culture medium. After that, purity of cells was approved by FCԑRI and CD117 antibodies, toluidine blue and Immunohistochemistry (IHC). Results: After 3 weeks continuous culturing, high purity of cells was found. CD117, CD34 expression and tryptase were 80.1, 76.89 and 87.9%, respectively by rat splenic supernatant, whereas 85.4, 83.07 and 82.1%, respectively with mouse splenic supernatants. Besides, rat spleen supernatant developed 91.4% and mouse splenocyte supernatant developed 89.7% mast cells based on surface markers. Conclusion: The data presented in this study indicated equal maturation and differentiation of bone marrow derived rat mast cells by using both spleen supernatants.

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