Kirshenbaum A S, Goff J P, Metcalfe D D
Mast Cell Physiology Section, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Immunology. 1989 Sep;68(1):120-5.
It has been reported that rare colonies of mast cells may be identified when human bone marrow cells are cultured in agar. Based on this observation, we attempted to culture mast cells from human bone marrow using a modified agar interphase culture. Metachromatically staining cells appearing in culture peaked in number by 2-3 weeks and consisted of basophil-like cells, mast-like cells and larger, granulated cells superficially resembling mast cells. The large cells, which constituted the majority of metachromatic cells, were berberine sulphate-positive but negative for histamine, chloroacetate esterase, and mast cell tryptase. These larger granulated cells were subsequently identified as macrophages by FACS analysis with Leu M3 and by electron microscopy. Berberine sulphate-staining of macrophages was not due to the de novo synthesis of heparin, as shown by analysis of radiolabelled proteoglycans from cultured cells. The macrophage metachromasia was eliminated with the use of agarose. The metachromatic and berberine sulphate-staining of cultured human macrophages was therefore considered to be due to phagocytosed agar. Recognition of this phenomenon will aid in the proper identification of human mast cells and basophils in cultures where agar or similar substances are present, and demonstrates that berberine sulphate-staining is not specific for heparin.
据报道，当人骨髓细胞在琼脂中培养时，可能会识别出罕见的肥大细胞集落。基于这一观察结果，我们尝试使用改良的琼脂界面培养法从人骨髓中培养肥大细胞。培养中出现的异染性染色细胞数量在2至3周时达到峰值，由嗜碱性粒细胞样细胞、肥大细胞样细胞以及表面类似肥大细胞的更大的颗粒细胞组成。构成大多数异染性细胞的大细胞对硫酸小檗碱呈阳性，但对组胺、氯乙酸酯酶和肥大细胞类胰蛋白酶呈阴性。随后通过使用抗Leu M3的荧光激活细胞分选分析（FACS）和电子显微镜鉴定这些较大的颗粒细胞为巨噬细胞。如对培养细胞中放射性标记蛋白聚糖的分析所示，巨噬细胞的硫酸小檗碱染色并非由于肝素的从头合成。使用琼脂糖可消除巨噬细胞的异染性。因此，培养的人巨噬细胞的异染性和硫酸小檗碱染色被认为是由于吞噬的琼脂所致。认识到这一现象将有助于在存在琼脂或类似物质的培养物中正确鉴定人肥大细胞和嗜碱性粒细胞，并表明硫酸小檗碱染色并非肝素所特有的。
