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Simple propagation method for resident macrophages by co-culture and subculture, and their isolation from various organs.
BMC Immunology ( IF 2.9 ) Pub Date : 2019-09-18 , DOI: 10.1186/s12865-019-0314-z
Kazushige Ogawa 1 , Mayu Tsurutani 1 , Aya Hashimoto 1 , Miharu Soeda 1
Affiliation  

BACKGROUND Resident macrophages (Mø) originating from yolk sac Mø and/or foetal monocytes colonise tissues/organs during embryonic development. They persist into adulthood by self-renewal at a steady state, independent of adult monocyte inputs, except for those in the intestines and dermis. Thus, many resident Mø can be propagated in vitro under optimal conditions; however, there are no specific in vitro culture methods available for the propagation of resident Mø from diverse tissues/organs. RESULTS We provided a simple method for propagating resident Mø derived from the liver, spleen, lung, and brain of ICR male mice by co-culture and subculture along with the propagation of other stromal cells of the respective organs in standard culture media and successfully demonstrated the propagation of resident Mø colonising these organs. We also proposed a simple method for segregating Mø from stromal cells according to their adhesive property on bacteriological Petri dishes, which enabled the collection of more than 97.6% of the resident Mø from each organ. Expression analyses of conventional Mø markers by flow cytometry showed similar expression patterns among the Mø collected from the organs. CONCLUSION This is the first study to clearly provide a practical Mø propagation method applicable to resident Mø of diverse tissues and organs. Thus, this novel practical Mø propagation method can offer broad applications for the use of resident Mø of diverse tissues and organs.

中文翻译:

通过共培养和亚培养简单的繁殖方法,将其从各种器官中分离出来。

背景技术从卵黄囊Mø和/或胎儿单核细胞起源的常驻巨噬细胞(Mø)在胚胎发育过程中定植在组织/器官中。它们稳定地通过自我更新而持续到成年,与成人单核细胞输入无关,除了肠道和真皮中的输入。因此,许多居民Mø可以在最佳条件下体外繁殖。但是,目前尚无特定的体外培养方法可用于从各种组织/器官繁殖驻地Mø。结果我们提供了一种简单的方法,可以通过共培养和继代培养以及在标准培养基中繁殖各个器官的其他基质细胞来繁殖来自ICR雄性小鼠的肝脏,脾脏,肺脏和大脑的驻地Mø,并成功地证明了这一点。居民Mø繁殖这些器官。我们还提出了一种根据基质细胞在细菌培养皿中的粘附特性从基质细胞中分离Mø的简单方法,该方法能够从每个器官中收集97.6%以上的驻留Mø。通过流式细胞术对常规Mø标记物的表达分析表明,从器官收集的Mø中具有相似的表达模式。结论这是第一个清楚地提供适用于各种组织和器官的Mø的实用Mø传播方法的研究。因此,这种新颖的实用的Mø传播方法可以为各种组织和器官的常驻Mø的使用提供广泛的应用。通过流式细胞术对常规Mø标记物的表达分析表明,从器官收集的Mø中具有相似的表达模式。结论这是第一个清楚地提供适用于各种组织和器官的Mø的实用Mø传播方法的研究。因此,这种新颖的实用的Mø传播方法可以为各种组织和器官的常驻Mø的使用提供广泛的应用。通过流式细胞术对常规Mø标记物的表达分析表明,从器官收集的Mø中具有相似的表达模式。结论这是第一个清楚地提供适用于各种组织和器官的Mø的实用Mø传播方法的研究。因此,这种新颖的实用的Mø传播方法可以为各种组织和器官的常驻Mø的使用提供广泛的应用。
更新日期:2020-04-22
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