Characterization of rat tissue cultured mast cells

Abstract

Twelve continuous rat tissue cultured mast cell (MC) lines were established by prolonged culture of rat peritoneal MC in the absence of added factors or feeder layers. Two of these lines, RCMC1 and RCMC2, have been briefly described previously, seven others are now also described. Both RCMC1 and RCMC2 lack a marker chromosomes present on RBL-CA10.7 cells. All lines were found to express the phenotype of mucosal MC as defined by alcian blue-positive and safranin O-negative staining, the presence of rat MC protease II and a low histamine content. When analyzed for high-(Fc epsilon RI) and low-affinity (Fc epsilon RL) receptors for IgE, the various lines yielded a variety of receptor patterns. Northern blot analysis of the RNA of RCMC1, RCMC2 and RBL-CA10.7 revealed that all three cell lines contained the same mRNA species for the alpha, beta and gamma subunits for Fc epsilon RI previously found in another rat basophilic leukemia cell line. Quantitation of the relative amounts of alpha, beta and gamma mRNA did not correlate with the expression of the relative amounts of Fc epsilon RI(alpha) in these cells. The relative amounts of mRNA for all these subunits of RCMC2 were equal or higher than those of RCMC1, suggesting that the low expression of Fc epsilon RI(alpha) on the former was a consequence of post-transcriptional events. Analysis of a RCMC1 clone over a 6-month period revealed changes in the expression of both Fc epsilon RI(alpha) and Fc epsilon RL.

Nitric oxide: a major determinant of mast cell phenotype and function

“…Rat cultured MC (RCMC), a gift from Drs A Froese and B. Chan, University of Manitoba were cultured as described previously (Chan et al 1990). Mouse bone marrow cultured MC (BMMC) were grown using the protocol of Vliagoftis (2002).…”

Selective antibody blockade of lymphocyte migration to mucosal sites and mast cell adhesion

“…Manitoba, Winnipeg, Canada [25] and maintained in RPMI-1640 (GIBCO-BRL) plus 10% FBS. The production of murine MAdCAM-1-transfected CHO cells has been previously described [12].…”

Differential IgE Receptor Development on Rat Tissue Cultured Mast Cells

“…Monospecific horse antirat IgE was a gift from Dr. K.A. Kelly, and rabbit antibodies specific for FceRL were prepared and tested as outlined before [8,13].…”

“…Re cently, receptor analysis of RCMC derived from rat peritoneal mast cells, but expressing the mucosal phenotype, revealed changes in receptor pattern upon continuous culture [7], Preliminary studies had sug- gested that these changes were inherent in the cells and did not involve changes in the cell population [8].…”

“…In the rat, the former are represented by peritoneal mast cells [3], while examples of the latter are intestinal mucosal mast cells [5], basophilic leukemia (RBL) cells [6], and tissue cultured mast cells (RCMC) recently estab lished in this laboratory [7,8]. It is now recognized that both mast cell types have their origin in the bone marrow [2], but the exact lineage relationship between them remains to be established.…”