Culture expansion of primary human nasal epithelial cells (NEC) isolated with a nasal scraping spoon

利用鼻刮匙分离原代人鼻上皮细胞(NEC)进行体外培养扩增

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Abstract

OBJECTIVE: To obtain high-purity nasal epithelial cells (NEC) while avoiding the irritation experienced by patients during nasal biopsies. METHODS: This prospective, observational study enrolled patients undergoing surgical treatment for nasal septum deviation. After general anaesthesia, a novel nasal scraping spoon was used to collect epithelial cells from the mid-part of the inferior turbinate. The cells were evenly plated on six-well plates coated with rat tail collagen. The morphology and growth of the cells were observed at different time-points using an inverted phase-contrast microscope. Immunofluorescent staining of cytokeratin 18 was used to identify NEC. Ki67 staining was used to check cell viability. RESULTS: This study collected samples from 19 patients during a short procedure. No postoperative complications were observed. Cell samples ranging from 8.31 × 10(5) to 2.04 × 10(6) cells/sample were obtained. The culture model was suitable for primary NEC culture as demonstrated by the faster proliferation (5-7 days). There was no fungal or bacterial contamination. Immunofluorescent staining confirmed the presence and proliferative activity of NEC in the cultures. CONCLUSION: A novel nasal scraping spoon provided an easy sampling method, avoided nasal injuries and psychological barriers to sampling and sufficient viable NEC to establish primary cultures.

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