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为了研究确定人巨细胞病毒（ＨＣＭＶ）体外感染Ｂａｌｂ／ｃ新生乳鼠原代培养脑神经细胞及其感染特征，制备和培养了新生乳鼠脑神经细胞并进行病毒感染。感染后每隔２～３天在倒置显微镜下观察活细胞培养物并摄影记录。在感染后第１～４周，取样进行苏木素－伊红（ＨＥ）染色及尼氏（Ｎｉｓｌ）染色。同时，用已知抗ＨＣＭＶ外膜蛋白的单克隆抗体进行免疫细胞化学染色，和地高辛标记的ＨＣＭＶ寡核苷酸特异性探针进行原位杂交，检测相应病毒核酸及胞内定位。结果在病毒感染后的第２周，受染细胞出现明显病变，表现为颗粒增多，折光性减弱。ＨＥ染色可见，受染神经细胞明显肿胀，胞浆及胞核均为嗜酸性，并可见到嗜酸性包涵体。Ｎｉｓｓｌ氏染色发现，胞核旁的尼氏小体变小，成粉末状，甚至消失。免疫细胞化学染色显示，受染神经细胞呈阳性反应。原位杂交结果证实，病毒核酸存在于受染细胞的胞浆及核内。这表明，ＨＣＭＶ能感染体外原代培养的新生乳鼠脑神经细胞，且病理特征与在人胚脑神经细胞观察到的基本模式一致。这种体外培养的细胞模型，可用于ＨＣＭＶ致中枢神经系统感染的研究。
Newborn neonatal rat brain neurons were prepared and cultured for viral infection in order to study the identification of primary cultured brain neurons infected with human cytomegalovirus (HCMV) in Balb / c neonatal rats and their infection characteristics. Live cell cultures were observed under an inverted microscope every 2-3 days after infection and photographed. Samples were taken for hematoxylin-eosin (HE) staining and Nisl staining 1 to 4 weeks after infection. At the same time, immunocytochemistry was performed with a monoclonal antibody known to be anti-HCMV outer membrane protein, and in situ hybridization was performed with digoxigenin-labeled HCMV oligonucleotide-specific probe to detect the corresponding viral nucleic acid and intracellular localization. Results in the first 2 weeks after virus infection, the infected cells showed obvious lesions, the performance of particles increased, refraction decreased. Hematoxylin-eosin staining showed that the infected neurons were obviously swollen with cytoplasm and nucleus being eosinophilic and eosinophilic inclusion bodies were seen. Nissl’s staining found that Nissl body next to the nucleus becomes smaller, into a powder, or even disappear. Immunocytochemical staining showed that the infected nerve cells were positive. In situ hybridization results confirmed that the virus nucleic acid is present in the cytoplasm and nucleus of infected cells. This indicates that HCMV can infect primary cultured neonatal rat brain neurons in vitro and that the pathological features are consistent with the basic patterns observed in human embryonic brain neurons. This in vitro cultured cell model can be used for HCMV-induced CNS infection.