Characterization of 'adult-type' mast cells derived from human bone marrow CD34+ cells cultured in the presence of stem cell factor and interleukin-6.: Interleukin-4 is not required for constitutive expression of CD54, FcεRIα and chymase, and CD13 expression is reduced during differentiation
Characterization of 'adult-type' mast cells derived from human bone marrow CD34+ cells cultured in the presence of stem cell factor and interleukin-6.: Interleukin-4 is not required for constitutive expression of CD54, FcεRIα and chymase, and CD13 expression is reduced during differentiation
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DOI：
10.1046/j.1365-2222.2002.01373.x
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发表时间：
2002-06-01
影响因子：
6.1
通讯作者：
Schwartz, LB
中科院分区：
文献类型：
--
作者：
Shimizu, Y;Sakai, K;Schwartz, LB
背景 体外衍生的人类肥大细胞表现出不同的特性，部分取决于祖细胞的来源。大多数研究使用胎儿肝脏、脐带血或外周血。目的比较重组人干细胞因子(rhSCF)和重组人白细胞介素6(rhIL-6)体外培养的骨髓和脐带血中CD34(+)祖细胞的人肥大细胞。方法与结果培养12周后，细胞几乎全部为肥大细胞，且胞质颗粒几乎全部含有类胰蛋白酶和糜蛋白酶(MCTC)。型），用酸性甲苯胺蓝进行异染染色。并在细胞表面表达CD117。培养两周后即可检测到类胰蛋白酶蛋白和mRNA。培养 4 周时检测到食糜酶 mRNA 和蛋白，但培养 2 周时未检测到。 12 周时，通过 ELISA 测量每个细胞的食糜酶含量。人骨髓来源的肥大细胞 (HBMMC) (5.6 +/- 0.9 pg) 显着高于 (P < 0.05) 脐带血来源的肥大细胞 (CBMC) (2.4 +/- 0.9 pg)，而组胺和类胰蛋白酶水平没有显着差异。在测试的簇名称中，CD29、CD49d、CD51 和 CD61 在 HBMMC 上强烈表达。 CD54和Fc epsilon RI α也被组成型表达。第 0 天时，大约一半的 CD34 分选细胞为 CD13(+)，并且随着肥大细胞成熟，这种情况逐渐减少。电镜观察显示12周龄HBMC具有许多分泌颗粒，其中含有球形电子致密核心，周围有电子透明空间，这与体内发育的未成熟MCTC细胞的报道一致。结论人骨髓CD34(-)祖细胞是肥大细胞祖细胞的丰富来源，能够在rhSCF和rhIL-6存在下表达成熟肥大细胞的颗粒和表面标志物。
Background In vitro-derived human mast cells exhibit different properties, depending in part on the source of progenitor cells. Most investigations ha e used fetal Liver, cord blood or peripheral blood. Few have used adult bone marrow.Objective Human mast cells derived in vitro from the CD34(+) progenitors in bone marrow and cord blood that had been cultured with recombinant human stem cell factor (rhSCF) and recombinant human interleukin-6 (rhIL-6) were compared.Methods and results After 12 weeks of culture, nearly all of the cells were mast cells, and nearly all of these had cytoplasmic granules containing both tryptase and chymase (MCTC type), stained metachromatically with acidic toluidine blue. and expressed CD117 on the cell surface. Both tryptase protein and mRNA were detected by two weeks of culture. Chymase mRNA and protein were detected at 4 weeks but not at 2 weeks of culture. By 12 weeks, chymase content per cell, measured by ELISA. was significantly higher (P < 0.05) in human bone marrow-derived mast cells (HBMMC) (5.6 +/- 0.9 pg) than in cord blood-derived mast cells (CBMC) (2.4 +/- 0.9 pg), whereas histamine and tryptase levels were not significantly different. Of the cluster designations tested, CD29, CD49d, CD51 and CD61 were strongly expressed on HBMMC. CD54 and Fc epsilon RI alpha also were expressed constitutively. Approximately half of CD34-sorted cells at day 0 were CD13(+) and this diminished as mast cell maturation occurred. Electron microscopy revealed that 12-week-old HBMMC had many secretory granules that contained spherical electron dense cores surrounded by electron lucent space, consistent with previous reports of immature MCTC cells developing in vivo.Conclusions CD34(-) progenitors of human bone marrow are a rich source of mast cell progenitors capable of expressing granule and surface markers of mature mast cells in the presence of rhSCF and rhIL-6.