Retinoic acid is a negative regulator for the differentiation of cord blood-derived human mast cell progenitors

Abstract

We examined the effects of retinoids on the human mast cell development using a serum-deprived culture system. When 10-week cultured mast cells derived from CD34+ cord blood cells were used as target cells, both all-trans retinoic acid (ATRA) and 9-cis RA inhibited the progeny generation under stimulation with stem cell factor (SCF) in a dose-dependent manner (the number of progeny grown by SCF plus RA at 10−7 mol/L was one tenth of the value obtained by SCF alone). The early steps in mast cell development appear to be less sensitive to RA according to the single CD34+c-kit+ cord blood cell culture study. The optimal concentration of RAs also reduced the histamine concentration in the cultured mast cells (3.00 ± 0.47 pg per cell in SCF alone, 1.44 ± 0.18 pg per cell in SCF+ATRA, and 1.41 ± 0.10 pg per cell in SCF+9-cis RA). RT-PCR analyses showed the expression of RAR, RARβ, RXR, and RXRβ messenger ribonucleic acid (mRNA) in 10-week cultured mast cells. The addition of an RAR-selective agonist at 10−10 mol/L to 10−7 mol/L decreased the number of mast cells grown in SCF, whereas an RXR-selective agonist at up to 10−8 mol/L was inactive. Among RAR subtype selective retinoids used at 10−9 mol/L to 10−7 mol/L, only the RAR agonist was equivalent to ATRA at 10−7 mol/L in its ability to inhibit mast cell growth. Conversely, the addition of excess concentrations of a RAR antagonist profoundly counteracted the retinoid-mediated suppressive effects. These results suggest that RA inhibits SCF-dependent differentiation of human mast cell progenitors through a specific receptor.

STI571 inhibits growth and adhesion of human mast cells in culture

“…3A). A similar cell cycle change was observed in ATRA, as described previously [23]. On the other hand, the addition of STI571 resulted in an emergence of cells with less than G1 amounts of DNA, as well as a decrease in the frequency of cells in the SϩG2/M phase on day 2.…”

“…Serum-deprived liquid cultures were carried out in 24-well culture plates (#3047; Becton Dickinson) using a modification of the technique described previously [22][23][24]. To examine the effects of STI571 on the SCF-dependent development of mast cells, we used 10-week cultured cells grown with 20 ng/mL of SCF from CD34 ϩ cord blood cells as target cells, because a large number of such cultured cells were positive for tryptase, as described previously [22,23].…”

“…Serum-deprived liquid cultures were carried out in 24-well culture plates (#3047; Becton Dickinson) using a modification of the technique described previously [22][23][24]. To examine the effects of STI571 on the SCF-dependent development of mast cells, we used 10-week cultured cells grown with 20 ng/mL of SCF from CD34 ϩ cord blood cells as target cells, because a large number of such cultured cells were positive for tryptase, as described previously [22,23]. In a doseϪresponse study, 10-week cultured cells (1ϫ10 5 ) were incubated for 2 weeks in 24-well culture plates containing 2 mL of ␣-medium supplemented with 1% bovine serum albumin (BSA), 300 g/mL fully ironsaturated human transferrin (ϳ98% pure, Sigma), 16 g/mL soybean lecithin (Sigma), 9.6 g/mL cholesterol (Nakalai Chemicals Ltd., Tokyo, Japan), 20 ng/mL of SCF with or without various concentrations of STI571.…”

“…Histamine and tryptase concentrations in the cell lysates obtained by the treatment of the cultured cells with 0.5% Nonidet P-40 were measured with a Histamine Radioimmunoassay (RIA) Kit (Immunotech), and a fluoroenzymeimmunoassay (UniCAP Tryptase, Pharmacia and Upjohn Diagnostics AB, Uppsala, Sweden), respectively, as described previously [22,23].…”

RNAseq studies reveal distinct transcriptional response to vitamin A deficiency in small intestine versus colon, uncovering novel vitamin A-regulated genes

“…This is in line with previous observations. Retinol and RA were shown to inhibit the growth and differentiation of human leukemic mast cells (HMC-1) (102), and the mast cells derived from peripheral blood or cord blood samples (103,104). Peritoneal mast cell numbers were also reported to be 1.5 times higher in VAD rat (105).…”