Elbert K J, Schäfer U F, Schäfers H J, Kim K J, Lee V H, Lehr C M
Department of Biopharmaceutics and Pharmaceutical Technology, Saarland University, Saarbrücken, Germany.
Pharm Res. 1999 May;16(5):601-8. doi: 10.1023/a:1018887501927.
To develop a cell culture model of human alveolar epithelial cells in primary culture for the in vitro study of pulmonary absorption and transport.
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.
In culture, the isolated cells spread into confluent monolayers, exhibiting peak transepithelial resistance of 2,180 +/- 62 ohms x cm2 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 Papp values ranging from 1.7 to 0.2 x 10(-8) cm/sec.
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.
建立人肺泡上皮细胞原代培养的细胞模型，用于肺部吸收和转运的体外研究。
通过酶处理和后续纯化从正常人远端肺组织分离出的II型肺细胞接种于纤连蛋白/胶原蛋白包被的聚酯滤膜插入物上，并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对细胞培养进行表征。
在培养过程中，分离的细胞铺展形成汇合单层，表现出2180±62欧姆·平方厘米的峰值跨上皮电阻和13.5±1.0毫伏的电位差（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型细胞组成，其间散布着一些II型细胞，从而很好地模拟了体内情况。发现亲水性大分子异硫氰酸荧光素标记的葡聚糖跨细胞单层的渗透率与其分子大小成反比，Papp值范围为1.7至0.2×10⁻⁸厘米/秒。
已建立人肺泡上皮细胞原代培养模型，该模型似乎是用于肺部药物递送和转运研究的有价值的体外模型。
