Kulka Marianna, Metcalfe Dean D
National Research Council Canada, Institute for Nutrisciences and Health, Charlottetown, Prince Edward, Island, Canada.
Curr Protoc Immunol. 2010 Aug;Chapter 7:Unit 7.25. doi: 10.1002/0471142735.im0725s90.
Located primarily in tissues, mast cells are one of the principal effector cells in allergic inflammation. Mast cells derive from mononuclear precursor cells which undergo their final phase of differentiation in the tissues. Mast cells express a unique set of proteases and display functional diversity depending on the tissue in which they differentiate--a phenomenon often referred to as mast cell heterogeneity. Enzymatic digestion and density centrifugation have often been used to isolate human mast cells from tissues such as lung and skin, frequently resulting in cells with low viability and purity. Here, we describe a protocol that combines gentle enzymatic digestion with positive selection techniques to isolate reasonably viable and substantially enriched preparations of tissue mast cells.
肥大细胞主要位于组织中，是过敏性炎症的主要效应细胞之一。肥大细胞来源于单核前体细胞，这些细胞在组织中经历其分化的最后阶段。肥大细胞表达一组独特的蛋白酶，并根据它们分化所在的组织表现出功能多样性——这种现象通常被称为肥大细胞异质性。酶消化和密度离心常常被用于从肺和皮肤等组织中分离人肥大细胞，这常常导致细胞活力和纯度较低。在此，我们描述了一种将温和的酶消化与阳性选择技术相结合的方案，以分离出具有相当活力且大量富集的组织肥大细胞制剂。
