A Research for the Development of Antiallergic-Drug with using Growing Differentiated Mast Cells in Culture System

本研究通过对肥大细胞体外培养体系的研究，获得了以下结果。培养的肥大细胞有利于抗过敏药物作用机制的研究或快速判断药物的疗效.分化肥大细胞的制备和培养：在含血清培养基中培养期间，分化良好的腹膜肥大细胞逐渐失去其含组胺的颗粒。然而，通过添加来自胎肝细胞的条件培养基或通过与胎肝细胞作为饲养层细胞共培养来保护消失。有些肥大细胞具有类似单个神经细胞的轴突。通过加入大量来自胎儿细胞的条件培养基，加速了这种形状的转变。IL-3依赖性肥大细胞（P-3细胞）也在来自WHEI细胞的条件培养基存在下培养。肥大细胞瘤P-815细胞在丁酸盐或糖皮质激素存在下生长。丁酸刺激肥大细胞的各种分化活性（组胺、乙酰肝素、肝素合成和粒细胞生成），而糖皮质激素仅刺激组氨酸脱羧酶的合成。2）培养肥大细胞生物活性物质的合成和分泌：DNP-蛔虫抗血清和化合物48/80活性寡聚体刺激肥大细胞分泌组胺和多种花生四烯酸代谢产物。脱颗粒肥大细胞在cAMP或糖皮质激素的作用下逐渐将其生物胺重组为颗粒。3）抗过敏药物能有效地抑制各种肥大细胞依赖于Ca^（2+）的分泌机制，并呈剂量依赖性。这些结果表明，培养的肥大细胞可用于研究和开发与肥大细胞功能相关的抗过敏药物。

We obtained the following results with a serious study how to grow the well-differentiated mast cells in vitro culture system. The cultured mast cells are beneficial to develop the mechanism of action of anti-allergic drugs or to judge their effectiveness quickly.1. Preparation and cultivation of differentiated mast cells: During cultivation in serum-containing medium, well-differentiated peritoneal mast cells gradually lost their histamine-containing granules. However, the disappearance was protected by addition of conditioned medium from, or by co-cultivation with fetal liver cells as a feeder layer cells. Some of grown mast cells have an axon like a single nerve cells. This transformation in shape was accelerated by adding large amount of conditioned medium from fetal cells. IL-3 dependent mast cells (P-3 cells) was also cultivated in the presence of conditioned medium from WHEI cells. Mastocytoma P-815 cells were grown in the presence of butyrate or glucocorticoid. Butyrate stimulated evry differentiation activities (histamine-, serotnin-, heparin-synthesis and granulopoiesis), while glucocorticoid only his-tidine decarboxylase synthesis.2) Cell activities of cultivated mast cells in the synthesis and the secretion of biological active compounds: DNP-ascaris antiserum and active oligomer of compound 48/80 stimulated the secretion of histamine and various arachidonic metabolites fron cultured mast cells. Also, degranulated mast cells gradually reconstituted their biological amines in granules by cAMP or glucocorticoid.3) Well-known antiallergic drugs effectively and dose-dependently suppressed the <Ca^(2+)> -dependent secretion mechanism of every types of cultured mast cells. These results show that the cultured mast cells appear to be available in the research and the development of anti-allergic drugs in associaton with the mast cell function.