Bischoff S C, Schwengberg S, Raab R, Manns M P
Department of Gastroenterology, Medical School of Hannover, Germany.
J Immunol. 1997 Dec 1;159(11):5560-7.
Culture of human mast cells (MC) in vitro has only been possible to date in the presence of 3T3 fibroblasts. The aim of the present study was to maintain freshly isolated human MC in culture without addition of feeder cells and to study their functional properties. We isolated cell suspensions containing 1 to 11% MC from human intestinal tissue and cultured them in standard medium. MC survived in culture for about 2 wk without cytokine supplementation and for several months with supplementation of medium with stem cell factor (SCF). SCF selectively supported MC survival, whereas the number of contaminating cells declined rapidly during culture. Most interestingly, we found that histamine and leukotriene release induced by IgE receptor cross-linking was substantially enhanced in cultured MC compared with that in MC stimulated directly after cell isolation. Cultured MC, but not freshly isolated MC, released mediators in response to SCF in a concentration-dependent fashion provided that the cells were cultured in SCF-free medium. These findings demonstrate that human MC isolated from intestinal tissue can be maintained in culture in vitro for several weeks. After culture they have different functional properties, which might resemble more closely the functional status of human intestinal MC in vivo than that of freshly isolated cells.
迄今为止,人类肥大细胞(MC)的体外培养只能在3T3成纤维细胞存在的情况下进行。本研究的目的是在不添加饲养细胞的情况下培养新鲜分离的人MC,并研究其功能特性。我们从人肠道组织中分离出含有1%至11%MC的细胞悬液,并在标准培养基中进行培养。在不添加细胞因子的情况下,MC在培养中存活约2周,在添加干细胞因子(SCF)的培养基中存活数月。SCF选择性地支持MC存活,而污染细胞的数量在培养过程中迅速下降。最有趣的是,我们发现与细胞分离后直接刺激的MC相比,培养的MC中由IgE受体交联诱导的组胺和白三烯释放显著增强。如果细胞在无SCF的培养基中培养,培养的MC而非新鲜分离的MC会以浓度依赖的方式对SCF释放介质。这些发现表明,从肠道组织分离的人MC可以在体外培养数周。培养后它们具有不同的功能特性,这可能比新鲜分离的细胞更接近体内人肠道MC的功能状态。