人鼻黏膜上皮细胞的原代培养及传代

Primary culture and subculture of human nasal mucosa epithelial cell

摘要:

目的探讨人鼻黏膜上皮细胞的原代培养及传代方法。方法取鼻内镜下行鼻中隔手术时切除的正常下鼻甲黏膜。以DMEM/F12培养基及KeratinocyteSFM（KSFM）培基先后培养及传代，同时培养鼻咽上皮细胞NP69作参照。于倒置显微镜下进行细胞形态学观察，以台盼蓝染色观察细胞活力，免疫细胞化学进行上皮细胞鉴定。结果鼻黏膜上皮细胞呈鹅卵石样生长，传代后形态一致性提高，细胞活力达88.1%，纯度达92.97%，免疫组织化学证实为上皮细胞。结论此方法培养的鼻黏膜上皮细胞活力及纯度高,并可进行一定程度的传代，是获得人鼻黏膜上皮细胞的一种有效方法。

关键词:鼻黏膜|上皮细胞|原代培养

Abstract:

ObjectiveTo explore a primary culture and subculture system of human nasal mucosa epithelial cell.MethodsNormal infraturbinal mucosa was obtained during endoscopic nasal septum operation. The tissue block was primarily cultured in DMEM/F12 and subcultured in KeratinocyteSFM (SFM) medium respectively. Meanwhile, nasopharyngeal epithelial cells NP69 were cultured as control. Cell morphology was observed under inverted microscope. Cell vitality was determined via Trypan blue staining, and cells were identified with immunohistochemical method.ResultsThe epithelial cells of nasal mucosa showed cobblestonelike growth. After passage, their morphological consistency was improved. The cell viability was 88.1% and the purity was up to 92.97%. Immunohistochemistry confirmed that they were epithelial cells.ConclusionThis method is effective to obtain human nasal epithelial cells, which have high activity and purity, and can produce subgeneration in a certain extent.

Key words:Nasal mucosa|Epithelial cell|Primary culture