Microglia are glial-immune cells that are essential for the function and survival of the central nervous system. Microglia not only protect neural tissues from immunological insults, but also play a critical role in neural development and repair. However, little is known about the biology of microglia in the cochlea, the auditory portion of the inner ear. In this study, we detected TMEM119 +, CD11b +, CD45 + and Iba1 + populations of cells in the rat cochlea, particularly in Rosenthal's canal, inner sulcus and stria vascularis. Next, we isolated and enriched the population of CD11b + cells from the cochlea and immortalized these cells with the 12S E1A gene of adenovirus in a replication-incompetent retroviral vector to derive a novel microglial cell line, designated Mocha (microglia of the cochlea). The resulting Mocha cells express a number of markers consistent with microglia and respond to lipopolysaccharide (LPS) stimulation by upregulation of genes (Cox2, ICAM-1, Il6r, Ccl2, Il13Ra and Il15Ra) as well as releasing cytokines (IL-1beta, IL-12, IL-13 and RANTES). As evidence of microglial function, Mocha cells phagocytose fluorescent beads at 37 °C, but not at 4 °C. The expression pattern of microglial markers in Mocha cells suggests that immortalization leads to a more primitive phenotype, a common phenomenon in immortalized cell lines. In summary, Mocha cells display key characteristics of microglia and are now available as a useful model system for the study of cochlear microglial behavior, both in vitro and in vivo.

小胶质细胞是神经胶质免疫细胞，对中枢神经系统的功能和生存至关重要。小胶质细胞不仅保护神经组织免受免疫损伤，而且在神经发育和修复中起关键作用。然而，对于耳蜗，内耳的听觉部分的小胶质细胞的生物学知之甚少。在这项研究中，我们检测到大鼠耳蜗，尤其是卢森塔尔管，内沟和血管纹中的TMEM119 +，CD11b +，CD45 +和Iba1 +细胞群体。接下来，我们从耳蜗中分离并富集了CD11b +细胞的群体，并在无复制能力的逆转录病毒载体中用腺病毒的12S E1A基因使这些细胞永生化，以衍生出一种新的小胶质细胞系，称为Mocha（耳蜗小胶质细胞）。产生的Mocha细胞表达许多与小胶质细胞一致的标记，并通过上调基因（Cox2，ICAM-1，Il6r，Ccl2，Il13Ra和Il15Ra）以及释放细胞因子（IL-1beta，IL）来响应脂多糖（LPS）刺激。 -12，IL-13和RANTES）。作为小胶质细胞功能的证据，Mocha细胞在37°C而不是4°C时吞噬了荧光珠。摩卡细胞中小胶质细胞标记物的表达模式表明永生化导致更原始的表型，这是永生化细胞系中的常见现象。总之，摩卡细胞显示出小胶质细胞的关键特征，现在可作为研究耳蜗小胶质细胞行为的有用模型系统 Il13Ra和Il15Ra）以及释放的细胞因子（IL-1beta，IL-12，IL-13和RANTES）。作为小胶质细胞功能的证据，Mocha细胞在37°C而不是4°C时吞噬了荧光珠。摩卡细胞中小胶质细胞标记物的表达模式表明永生化导致更原始的表型，这是永生化细胞系中的常见现象。总之，摩卡细胞显示出小胶质细胞的关键特征，现在可作为研究耳蜗小胶质细胞行为的有用模型系统 Il13Ra和Il15Ra）以及释放的细胞因子（IL-1beta，IL-12，IL-13和RANTES）。作为小胶质细胞功能的证据，Mocha细胞在37°C而不是4°C时吞噬了荧光珠。摩卡细胞中小胶质细胞标记物的表达模式表明永生化导致更原始的表型，这是永生化细胞系中的常见现象。总之，摩卡细胞显示出小胶质细胞的关键特征，现在可作为研究耳蜗小胶质细胞行为的有用模型系统 永生细胞系中的常见现象。总之，摩卡细胞显示出小胶质细胞的关键特征，现在可作为研究耳蜗小胶质细胞行为的有用模型系统 永生细胞系中的常见现象。总之，摩卡细胞显示出小胶质细胞的关键特征，现在可作为研究耳蜗小胶质细胞行为的有用模型系统体外和体内。