Institute of Pharmacology, Pharmacy and Toxicology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 15, 04103 Leipzig, Germany.
BMC Vet Res. 2011 Jun 7;7:26. doi: 10.1186/1746-6148-7-26.
Horses develop recurrent airway obstruction (RAO) that resembles human bronchial asthma. Differentiated primary equine bronchial epithelial cells (EBEC) in culture that closely mimic the airway cells in vivo would be useful to investigate the contribution of bronchial epithelium in inflammation of airway diseases. However, because isolation and characterization of EBEC cultures has been limited, we modified and optimized techniques of generating and culturing EBECs from healthy horses to mimic in vivo conditions.
Large numbers of EBEC were obtained by trypsin digestion and successfully grown for up to 2 passages with or without serum. However, serum or ultroser G proved to be essential for EBEC differentiation on membrane inserts at ALI. A pseudo-stratified muco-ciliary epithelium with basal cells was observed at differentiation. Further, transepithelial resistance (TEER) was more consistent and higher in P1 cultures compared to P0 cultures while ciliation was delayed in P1 cultures.
This study provides an efficient method for obtaining a high-yield of EBECs and for generating highly differentiated cultures. These EBEC cultures can be used to study the formation of tight junction or to identify epithelial-derived inflammatory factors that contribute to lung diseases such as asthma.
马会发生类似于人类支气管哮喘的复发性气道阻塞（RAO）。在培养中分化的原发性马支气管上皮细胞（EBEC）能够非常近似地模拟体内气道细胞，这对于研究支气管上皮在气道疾病炎症中的作用非常有用。然而，由于 EBEC 培养物的分离和鉴定受到限制，我们对从健康马匹中生成和培养 EBEC 的技术进行了改进和优化，以模拟体内条件。
通过胰蛋白酶消化获得了大量的 EBEC，并成功地在有无血清的情况下培养了多达 2 代。然而，血清或 ultroser G 对于在 ALI 上的 EBEC 分化是必需的。在分化过程中观察到具有基底细胞的假复层纤毛柱状上皮。此外，与 P0 培养物相比，P1 培养物的跨上皮电阻（TEER）更加一致且更高，而纤毛化在 P1 培养物中延迟。
本研究提供了一种高效获取大量 EBEC 和生成高度分化培养物的方法。这些 EBEC 培养物可用于研究紧密连接的形成，或鉴定上皮衍生的炎症因子，这些因子有助于哮喘等肺部疾病的发生。
