ESTABLISHMENT OF 2 DOG MASTOCYTOMA CELL-LINES IN CONTINUOUS CULTURE
ESTABLISHMENT OF 2 DOG MASTOCYTOMA CELL-LINES IN CONTINUOUS CULTURE
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DOI：
10.1165/ajrcmb/3.5.413
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发表时间：
1990-11-01
影响因子：
6.4
通讯作者：
GOLD, WM
中科院分区：
文献类型：
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作者：
DEVINNEY, R;GOLD, WM
我们之前已经确定了狗肥大细胞瘤细胞在裸鼠体内繁殖的特征。我们已经在连续培养中建立了其中两个品系(C和C2)。新鲜分离的肥大细胞瘤细胞在Dulbecco‘s改良Eagle’s培养液(DME)-HI6+50%Ham‘s F12+组氨酸和5%过敏性犬血清(ADS)中培养。细胞每3d喂养一次，每周传代一次。生长通过细胞计数进行评估。细胞生长以5%ADS的培养效果最好。细胞在ADS中悬浮生长，传代55次，倍增时间为37.4±18.7h(平均±1SD；n=15)。细胞贴壁于组织培养膜上，传代26次，倍增时间49.3±12.5h(n=13)。形态和功能特征与在裸鼠体内繁殖的细胞中描述的特征没有变化。C_1的组胺含量为0.46±0.18 pg/cell(n=12)，Cs的组胺含量为0.07±0.04 pg/cell(n=6)。两个品系都含有中性蛋白酶类胰蛋白酶，而Cz含有凝乳酶。钙离子载体A23187或豚草抗原可引起两种细胞释放浓度依赖的组胺。C和C产生前列腺素D，与A23187反应。我们的结论是，狗肥大细胞瘤细胞可以在连续培养中建立，从而为研究肥大细胞生物学提供了一个系统，包括生长和发育。肥大细胞被认为在多种疾病中发挥重要作用，包括哮喘、间质纤维化和肿瘤，以及炎症、代谢和修复反应(1-4)。它们产生各种颗粒相关和新形成的介质，以响应免疫和非免疫刺激。直到最近，还没有纯培养的人或狗的肥大细胞的制备，以前的研究依赖于啮齿动物肥大细胞(5-8)，或急性分离的人肺肥大细胞系(9，10)。
We have previously characterized dog mastocytoma cells propagated in nude mice. We have established two of these lines (C, and C2) in continuous culture. Freshly disaggregated mastocytoma cells were cultured in Dulbecco's modified Eagle's medium (DME)-HI6 mixed with 50% Ham's F12 and supplemented with histidine and 5% allergic dog serum (ADS). Cells were fed every 3 d and passaged weekly. Growth was assessed by cell count. Cell growth was best supported by culture in 5% ADS. C, cells grow in suspension in ADS and have been passaged 55 times with a doubling time of 37.4±18.7 h (mean±1 SD; n= 15). C, cells adhere to tissue culture plastic in ADS and have been passaged 26 times with a doubling time of 49.3±12.5 h (n= 13). Morphologic and functional characteristics are unchanged from those described in cells propagated in nude mice. Histamine content for C, is 0.46±0.18 pg/cell (n= 12) and 0.07±0.04 pglcell (n= 6) for Cs. Both lines contain the neutral protease tryptase and Cz contains chymase. Calcium ionophore A23187 or ragweed antigen caused concentration-dependent histamine release from both cell lines. C, and C, generate prostaglandin D, in response to A23187. We conclude that dog mastocytoma cells can be established in continuous culture, thus providing a system for studying mast cell biology, including growth and development.Mast cells are thought to play an important role in a variety of diseases, including asthma, interstitial fibrosis, and neoplasia, as well as inflammatory, metabolic, and reparative reactions (1-4). They produce a variety of granule-associated and newly formed mediators in response to both immunologic and nonimmunologic stimuli. Until recently, no preparation of pure cultured human or dog mast cells was available, and previous studies have relied on rodent mast cells (5-8), or on acutely isolated human lung mast cell lines (9, 10).