In vitro expansion of human and mouse mast cells by the ligand for c-ki

肥大细胞来源于多能造血干细胞，并分为至少两个表型不同的亚群；结缔组织型肥大细胞（CTMC）和粘膜肥大细胞（MMC）。当用IL-3培养正常小鼠骨髓细胞时，产生大量肥大细胞，相当于MMC亚类的组织培养物。从小鼠腹膜呼叫中纯化的成熟CTMC可以在IL-3和IL-4存在下增殖。干细胞因子（SCF）是c-kit酪氨酸激酶受体的配体，被证明与多种细胞因子（包括IL-3、IL-6和G-CSF）协同作用于早期造血祖细胞的发育。虽然SCF单独不能从骨髓谱系阴性细胞中诱导肥大细胞，但它与IL-3联合可刺激肥大细胞的产生。SCF、IL-3均不能诱导纯化成熟CTMC的肥大细胞集落，但SCF与IL-3、IL-4或GM-CSF在体外对小鼠CTMC的增殖有显著的协同作用。抗c-kit受体抗体（ACK2）能完全阻断SCF的活性，但不能阻断IL-3对CTMC生长的影响。当我们在SCF存在下培养从脐带血或骨髓细胞中纯化的人CD34^+细胞时，在培养14天后首次检测到人肥大细胞，并在培养50天后急剧增加。脐带血CD34^+细胞培养60天后，SCF、SCF+IL-6或SCF+IL-11培养的细胞90%以上为肥大细胞。IL-3和GM-CSF显著抑制scf依赖性肥大细胞的增殖。

Mast cell are derived from multipotential hematopoietic stem cells and classified into at least two phenotypically distinct subpopulations; connective tissue-type mast cells (CTMC) and mucosal mast cells (MMC). When normal mouse bone marrow cells were cultured with IL-3, a large number of mast cells ,which were tissue culture equivalents of the MMC subclass, were developed. Mature CTMC purified from mouse peritoneal calls could proliferate in the presence of both IL-3 and IL-4. Stem cell factor (SCF), the ligand for the c-kit tyrosine kinase receptor, demonstrated to act synergistically with a variety of cytokines including IL-3, IL-6, and G-CSF on the development of early hematopoietic progenitors. Although SCF alone failed to induce mast cells from bone marrow lineage negative cells, it stimulated mast cell production in combination with IL-3. Either SCF, IL-3 alone could not induce mast cell colonies from purified mature CTMC, but SCF revealed significant cooperative actions with IL-3, IL-4 or GM-CSF on the proliferation of mouse CTMC in vitro. The anti-c-kit receptor antibody (ACK2) completely blocked the activity of SCF but not of IL-3 on the growth of CTMC. When we cultured human CD34^+ cells purified from cord blood or bone marrow cells in the presence of SCF, human mast cells first detected on 14 days of incubation and increased dramatically after 50 days of culture. More than 90% of the cells in culture with SCF,SCF+IL-6 or SCF+IL-11 were mast cells after 60 days of culture of cord blood CD34^+ cells. IL-3 and GM-CSF significantly inhibited SCF-dependent mast cell proliferation.