Takagi M, Nakahata T, Kubo T, Shiohara M, Koike K, Miyajima A, Arai K, Nishikawa S, Zsebo K M, Komiyama A
Department of Pediatrics, Shinshu University School of Medicine, Matsumoto, Japan.
J Immunol. 1992 Jun 1;148(11):3446-53.
The proliferative capacity of mouse connective tissue-type mast cells (CTMC) was analyzed by using a newly discovered c-kit ligand, termed stem cell factor (SCF). More than 90% of CTMC in the peritoneal cavity responded to recombinant rat SCF (rrSCF) and were able to give rise to pure mast cell colonies in methylcellulose culture. Serial observation (mapping) of growth of individual CTMC in culture containing rrSCF confirmed their striking proliferative ability. No serum but accessory cells (non-CTMC cells) in the peritoneal population were required for the clonal growth of CTMC induced by rrSCF in our methylcellulose culture of whole peritoneal cells. The rrSCF-induced mast cell colony formation from peritoneal CTMC was completely inhibited by the addition of anti-c-kit antibody, which can block the binding of SCF to c-kit, to the culture. When IL-3 was combined with rrSCF, mast cell colonies dramatically increased in size. Mapping studies revealed that the combination of the two factors augmented the proliferative rate of CTMC. Approximately 60% of the constituent cells of the mast cell colonies which were formed from peritoneal CTMC in the culture containing rrSCF alone were stained with berberine sulfate, which is a characteristic of CTMC. However, most mast cells which were induced by rrSCF+IL-3 from peritoneal CTMC contained berberine(-)-safranin(-)-Alcian blue(+) granules. Although IL-4 exhibited little synergism with rrSCF in the induction of CTMC proliferation, the addition of IL-4 to the culture containing rrSCF+IL-3 resulted in an increase in mast cells which retained CTMC characteristics.
通过使用新发现的被称为干细胞因子（SCF）的c-kit配体，分析了小鼠结缔组织型肥大细胞（CTMC）的增殖能力。腹腔中超过90%的CTMC对重组大鼠SCF（rrSCF）有反应，并能够在甲基纤维素培养中形成纯肥大细胞集落。对含rrSCF培养物中单个CTMC生长的连续观察（绘图）证实了它们显著的增殖能力。在我们对全腹腔细胞进行的甲基纤维素培养中，rrSCF诱导CTMC克隆生长不需要血清，但需要腹腔细胞群体中的辅助细胞（非CTMC细胞）。向培养物中添加可阻断SCF与c-kit结合的抗c-kit抗体，可完全抑制rrSCF诱导的腹腔CTMC肥大细胞集落形成。当IL-3与rrSCF联合使用时，肥大细胞集落的大小显著增加。绘图研究表明，这两种因子的联合增强了CTMC的增殖速率。在仅含rrSCF的培养物中，由腹腔CTMC形成的肥大细胞集落中约60%的组成细胞被硫酸小檗碱染色，这是CTMC的一个特征。然而，rrSCF+IL-3诱导的腹腔CTMC产生的大多数肥大细胞含有小檗碱（-）-番红（-）-阿尔辛蓝（+）颗粒。尽管IL-4在诱导CTMC增殖方面与rrSCF几乎没有协同作用，但在含rrSCF+IL-3的培养物中添加IL-4会导致保留CTMC特征的肥大细胞数量增加。
