Monolayers of human alveolar epithelial cells in primary culture for pulmonary absorption and transport studies
Monolayers of human alveolar epithelial cells in primary culture for pulmonary absorption and transport studies
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DOI：
10.1023/a:1018887501927
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发表时间：
1999-05-01
影响因子：
3.7
通讯作者：
Lehr, CM
中科院分区：
文献类型：
--
作者：
Elbert, KJ;Schäfer, UF;Lehr, CM
目的.目的建立人肺泡上皮细胞体外培养模型，用于体外研究肺吸收和转运。将通过酶处理和随后的纯化从正常人远端肺组织分离的II型肺细胞铺在纤连蛋白/胶原包被的聚酯过滤器插入物上，并使用低血清生长培养基培养。通过生物电测量、细胞特异性凝集素结合、细胞连接的免疫组织化学检测以及不同分子量葡聚糖的跨上皮转运评估来表征细胞培养物。在培养中，分离的细胞扩散成融合的单层，表现出2，180 +/- 62 Omega x cm（2）的峰值跨上皮电阻和13.5 +/- 1.0 mV的电位差（n = 30-48），并形成紧密连接和桥粒。通过凝集素结合评估，细胞单层主要由I型细胞和一些散布的II型细胞组成，因此很好地模拟了体内情况。发现亲水性大分子FITC-葡聚糖穿过细胞单层的渗透性与其分子大小呈负相关，P-app值范围为1.7至0.2 x 10（-8）cm/sec。建立了人肺泡上皮细胞原代培养模型，为研究药物在肺内的释放和转运提供了一个有价值的体外模型。
Purpose. To develop a cell culture model of human alveolar epithelial cells in primary culture for the in vitro study of pulmonary absorption and transport.Methods. Type II pneumocytes isolated from normal human distal lung tissue by enzyme treatment and subsequent purification were plated on fibronectin/collagen coated polyester filter inserts, and cultured using a low-serum growth medium. Characterization of the cell culture was achieved by bioelectric measurements, cell-specific lectin binding, immunohistochemical detection of cell junctions, and by assessment of transepithelial transport of dextrans of varying molecular weights.Results. In culture, the isolated cells spread into confluent monolayers, exhibiting peak transepithelial resistance of 2,180 +/- 62 Omega x cm(2) and potential difference of 13.5 +/- 1.0 mV (n = 30-48), and developing tight junctions as well as desmosomes. As assessed by lectin-binding, the cell monolayers consisted of mainly type I cells with some interspersed type II cells, thus well mimicking the situation in vivo. The permeability of hydrophilic macromolecular FITC-dextrans across the cell monolayer was found to be inversely related to their molecular size, with P-app values ranging from 1.7 to 0.2 x 10(-8) cm/sec.Conclusions. A primary cell culture model of human alveolar epithelial cells has been established, which appears to be a valuable in vitro model for pulmonary drug delivery and transport studies.