Preparation of bone marrow-derived mast cells capable of producing high levels of histamine comparable to mature mast cells in vivo 9406

Abstract Description Mast cells are the crucial immune cells responsible for the production of histamine and diverse cytokines. Immature mast cells were prepared conventionally by culturing bone marrow cells (BMC) for culturing using low concentrations of IL-3 (5 ng/ml) and SCF (5 ng/ml) to produce the histamine level much lower than that of mature mast cells in vivo. The cells of rat bone marrow derived mast cells (rBMMCs) were studied for histamine production by culturing with high cytokine concentrations (IL-3: 50 ng/ml, SCF: 50 ng/ml) and high cell densities (5 x 105/ml). The cells were applied to transcriptome, real time PCR, and ELISA. The cells were analyzed for the effect of LPS on histamine production. rBMMCs cultured in conditions were measured to produce histamine reaching up to 25 ng/cell, comparable to the mature mast cells. The cells were observed to produce various cytokines and receptors in transcriptome analysis, The cells were analyzed to express the increase by 4.2-folds in HDC, by 870-folds in HRH1, by 2.7-fold in TLR4, and by 27-fold in TLR7 expression compared to non-cultured BMC in real time PCR. The histamine production of cells was increased by 2 with LPS. The cells were to produce TNF-α affected by cytokine concentration, measured via ELISA. These findings indicated that rat BMMCs are the good model for mast cell in vitro to produce the high level histamine comparable to mature mast cells in vivo, to respond to LPS and to exhibit elevated TLR4 and TNF-α production. Funding Sources not available Topic Categories Immediate Hypersensitivity, Asthma, and Allergic Responses (HYP)