Growth and differentiation of human nasal epithelial cells in culture. Serum-free, hormone-supplemented medium and proteoglycan synthesis.
Growth and differentiation of human nasal epithelial cells in culture. Serum-free, hormone-supplemented medium and proteoglycan synthesis.
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人鼻上皮细胞在培养物中的生长和分化。
DOI：
10.1164/arrd.1985.132.2.311
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Boucher,R
中科院分区：
文献类型：
--
作者：
Wu,R;Yankaskas,J;Cheng,E;Knowles,MR;Boucher,R
Ham 'sF_（12）培养基中添加胰岛素（Ins）、转铁蛋白（Tf）、表皮生长因子（EGF）、氢化可的松（HC）、T_3、霍乱毒素（CT）和牛下丘脑提取物（BHE），可培养人鼻上皮细胞（HNE）。用蛋白酶从新鲜切除的鼻息肉或鼻甲中分离HNE细胞。原代HNE细胞的集落形成效率约为5%。生长研究表明Ins、BHE和CT对生长是必需的，HC、EGF、Tf和T3也对生长有刺激作用。在这种无血清、添加酵母菌的培养基中的生长速率为24小时/群体倍增。可获得多达20个群体倍增和3代分离的HNE细胞。向该培养基中添加血清抑制上皮细胞生长。维生素A对细胞生长没有明显的影响，但诱导细胞形态特征的改变。培养细胞的上皮性质证实了阳性染色与抗人角蛋白抗体，超微结构的研究，并通过形成柱状，纤毛上皮细胞在裸露的气管移植重新填充这些培养的HNE细胞。对培养的HNE细胞分泌的糖蛋白（标记有3 H-氨基葡萄糖和/或35 S-硫酸盐）的生化分析不能证明培养物中的粘蛋白样糖蛋白的分泌。相反，培养细胞的主要分泌产物是透明质酸和硫酸乙酰肝素。这些结果与形态学观察结果一致，即培养细胞中没有粘液分泌颗粒。在高细胞密度培养物中观察到圆顶形成。我们的结论是，HNE细胞可以培养在定义明确的培养基。如圆顶的形成所示，这些细胞可用于体外离子转运研究。然而，粘蛋白合成的研究可能需要进一步的区分。
Ham's F12 medium supplemented with insulin (Ins), transferrin (Tf), epidermal growth factor (EGF), hydrocortisone (HC), T3, cholera toxin (CT), and bovine hypothalamus extract (BHE) was developed forin vitrogrowth of human nasal epithelial (HNE) cells. The HNE cells were dissociated from freshly excised nasal polyps or turbinates with protease. Colony-forming efficiency of primary HNE cells was approximately 5%. Growth studies showed Ins, BHE, and CT were essential for growth; HC, EGF, Tf, and T3were also stimulatory for growth. The growth rate in this serum-free, hormone-supplemented medium was 24 h per population doubling. Up to 20 population doublings and 3 passages of dissociated HNE cells could be achieved. Addition of serum to this culture medium inhibited epithelial cell growth. Vitamin A had no apparent effect on cell growth but induced an alteration in the morphologic characteristics of the cell. The epithelial nature of cultured cells was confirmed by positive staining with antihuman keratin antibody, ultrastructural studies, and by formation of a columnar, ciliated epithelium in denuded tracheal grafts repopulated by these cultured HNE cells. Biochemical analyses of glycoproteins (labeled with3H-glucosamine and/or35S-sulfate) secreted by cultured HNE cells were unable to demonstrate the secretion of mucinlike glycoproteins in culture. Instead, major secretory products of cultured cells were hyaluronate and heparan sulfate. These results were in agreement with morphologic observations that showed no mucus-secreting granules in cultured cells. Dome formation was observed in high cell density cultures. We conclude that HNE cells can be cultured in well-defined culture media. As indicated by formation of domes, these cells may be useful forin vitroion transport studies. Further differentiation, however, may be required for studies of mucin synthesis.