Niles R, Kim K C, Hyman B, Christensen T, Wasano K, Brody J
Department of Biochemistry, Boston University School of Medicine, Massachusetts.
In Vitro Cell Dev Biol. 1988 May;24(5):457-63. doi: 10.1007/BF02628498.
Studies on the regulation of differentiation in airway epithelial cells have been hampered by the lack of cell culture systems that differentiate in vitro. One such system that does exhibit differentiation is hamster tracheal epithelial cells (HTE). A major problem with this system, however, is that at the time cells differentiate, they lyze the collagen gel upon which they grow, resulting in termination of the culture. Here we report that by growing the HTE cells at 32 degrees instead of 37 degrees C we can totally prevent lysis of the collagen gel. Cells grown at this lower temperature maintain their differentiated phenotype as evidenced by abundant mucus granules and the secretion of authentic mucus glycoproteins into the culture media. We have also developed a method for subculturing the primary cells which allows growth and differentiation in secondary culture. The HTE cells were capable of being passaged at least three times and did not become transformed as judged by their inability to grow in soft agar and to produce tumors in syngeneic animals. This improved HTE cell culture system will allow detailed studies on the mechanisms which regulate growth, differentiation, and mucus secretion in surface airway epithelial cells.
气道上皮细胞分化调控的研究因缺乏能在体外分化的细胞培养系统而受阻。一种确实能表现出分化的系统是仓鼠气管上皮细胞（HTE）。然而，该系统的一个主要问题是，当细胞分化时，它们会溶解其生长所依赖的胶原凝胶，导致培养终止。在此我们报告，通过将HTE细胞在32摄氏度而非37摄氏度下培养，我们能够完全防止胶原凝胶的溶解。在这个较低温度下生长的细胞保持其分化表型，大量黏液颗粒以及向培养基中分泌真正的黏液糖蛋白证明了这一点。我们还开发了一种原代细胞传代培养的方法，该方法允许在二次培养中生长和分化。HTE细胞能够传代至少三次，并且根据它们在软琼脂中无法生长以及在同基因动物中无法产生肿瘤来判断，它们没有发生转化。这种改良的HTE细胞培养系统将允许对调节气道表面上皮细胞生长、分化和黏液分泌的机制进行详细研究。
