Dvorak A M, Furitsu T, Estrella P, Ishizaka T
Department of Pathology, Beth Israel Hospital, Boston, Massachusetts.
Am J Pathol. 1991 Dec;139(6):1309-18.
Culture systems designed to maintain or develop human mast cells have proved difficult, yet these systems would provide valuable resources for future investigations of human mast cell biology. Cocultures of either isolated mature human lung mast cells (Levi-Schaffer et al., J Immunol 1987, 139:494-500) or human cord blood mononuclear cells (Furitsu, Proc Natl Acad Sci USA 1989, 86:10039-10043) with 3T3 embryonic mouse skin fibroblasts have implicated fibroblasts as an important factor in the successful maintenance and development of human mast cells in vitro. The authors cultured isolated, mature human lung mast cells either with or without 3T3 cells for 1 month and examined their ultrastructural phenotype. Mast cell viability in each circumstance was equivalent, but mast cell yield was improved in the presence of 3T3 cells. The ultrastructural phenotype was identical in both culture systems. Mast cells were shown to maintain the phenotype of their in vivo lung counterparts (ie, scroll granules predominanted, and numerous lipid bodies were present). This ultrastructural phenotype differs from that of mast cells that develop in cocultures of human cord blood cells and 3T3 cells, where developing mast cells with crystalline granules and few lipid bodies prevail, a phenotype much like that of human skin mast cells in vivo (Furitsu, Proc Natl Acad Sci USA 1989, 86:10039-10043).
旨在维持或培养人肥大细胞的培养系统已被证明颇具难度,然而这些系统将为未来人类肥大细胞生物学研究提供宝贵资源。分离出的成熟人肺肥大细胞(Levi-Schaffer等人,《免疫学杂志》1987年,139:494 - 500)或人脐血单个核细胞(Furitsu,《美国国家科学院院刊》1989年,86:10039 - 10043)与3T3胚胎小鼠皮肤成纤维细胞的共培养表明,成纤维细胞是体外成功维持和培养人肥大细胞的一个重要因素。作者将分离出的成熟人肺肥大细胞与3T3细胞一起或不与3T3细胞一起培养1个月,并检查其超微结构表型。每种情况下肥大细胞的活力相当,但在有3T3细胞存在时肥大细胞产量有所提高。两种培养系统中的超微结构表型相同。肥大细胞显示出维持其体内肺对应细胞的表型(即卷曲颗粒占主导,且存在大量脂质体)。这种超微结构表型不同于在人脐血细胞与3T3细胞共培养中发育的肥大细胞,后者以具有结晶颗粒且脂质体较少的发育中肥大细胞为主,这种表型很像体内人皮肤肥大细胞的表型(Furitsu,《美国国家科学院院刊》1989年,86:10039 - 10043)。