Department of Medicine, Harvard Medical School, Boston, Mass, USA.
J Allergy Clin Immunol. 2011 Mar;127(3):815-22.e1-5. doi: 10.1016/j.jaci.2010.12.1101. Epub 2011 Feb 1.
Studies of human mast cells (MCs) are constrained by the paucity of functional cell lines, the expense of maintaining MCs in culture, and technical complexities.
We derived and characterized a human MC line that arose spontaneously from a culture of nontransformed hematopoietic progenitor cells.
CD34(+) enriched mononuclear cells derived from a donor with aspirin-exacerbated respiratory disease were cultured for 8 weeks with stem cell factor and IL-6 and with IL-3 for the first week only. The cells (termed LUVA cells) survived and proliferated without further addition of any growth factors and have been maintained in culture for approximately 2 years.
LUVA cells possess metachromatic cytoplasmic granules that are immunoreactive for tryptase, cathepsin G, and carboxypeptidase A3. They express transcripts encoding FcεRI, c-kit, chymase, tryptase, histidine decarboxylase, carboxypeptidase A3, and the type 1 receptor for cysteinyl leukotrienes. Flow cytometry confirmed uniform expression of FcεRI, c-kit, and FcγRII. FcεRI cross-linkage induced the release of β-hexosaminidase, prostaglandin D(2), thromboxane A(2), and macrophage inflammatory protein 1β. Immortalization was not associated with either a known genomic mutation of c-kit in the donor or a somatic mutation of c-kit within the cells, and it was not associated with c-kit autophosphorylation.
LUVA cells are an immortalized human MC line that can be maintained without stem cell factor and display high levels of normally signaling c-kit and FcεRI. These cells will prove valuable for functional human MC studies.
人类肥大细胞（MCs）的研究受到功能细胞系缺乏、MCs 在培养中维持成本高以及技术复杂性的限制。
我们从非转化造血祖细胞培养物中自发衍生并表征了一种人类 MC 系。
从患有阿司匹林加重的呼吸道疾病的供体中分离出 CD34+富集的单核细胞，并用干细胞因子和白细胞介素-6 培养 8 周，仅在第一周用白细胞介素-3 培养。这些细胞（称为 LUVA 细胞）无需添加任何生长因子即可存活和增殖，并已在培养中维持了大约 2 年。
LUVA 细胞具有嗜碱性细胞质颗粒，这些颗粒对胰蛋白酶、组织蛋白酶 G 和羧肽酶 A3 具有免疫反应性。它们表达编码 FcεRI、c-kit、糜蛋白酶、胰蛋白酶、组氨酸脱羧酶、羧肽酶 A3 和半胱氨酰白三烯 1 型受体的转录本。流式细胞术证实了 FcεRI、c-kit 和 FcγRII 的均匀表达。FcεRI 交联诱导β-己糖胺酶、前列腺素 D(2)、血栓烷 A(2)和巨噬细胞炎性蛋白 1β 的释放。永生化与供体中 c-kit 的已知基因组突变或细胞内 c-kit 的体细胞突变无关，也与 c-kit 自身磷酸化无关。
LUVA 细胞是一种永生化的人类 MC 系，可以在没有干细胞因子的情况下维持，并且显示出高水平的正常信号传导 c-kit 和 FcεRI。这些细胞将对功能性人类 MC 研究非常有价值。
