Marine Biomedicine and Environmental Science Center, Medical University of South Carolina, Hollings Marine Laboratory, Charleston, SC 29412, USA.
Comp Biochem Physiol C Toxicol Pharmacol. 2012 Jan;155(1):136-42. doi: 10.1016/j.cbpc.2011.04.002. Epub 2011 Apr 9.
Current models for in vitro studies of tissue function and physiology, including responses to hypoxia or environmental toxins, are limited and rely heavily on standard 2-dimensional (2-D) cultures with immortalized murine or human cell lines. To develop a new more powerful model system, we have pursued methods to establish and expand cultures of primary lung cell types and reconstituted tissues from marine mammals. What little is known about the physiology of the deep-sea diving pygmy sperm whale (PSW), Kogia breviceps, comes primarily from stranding events that occur along the coast of the southeastern United States. Thus, development of a method for preserving live tissues and retrieving live cells from deceased stranded individuals was initiated. This report documents successful cryopreservation of PSW lung tissue. We established in vitro cultures of primary lung cell types from tissue fragments that had been cryopreserved several months earlier at the stranding event. Dissociation of cryopreserved lung tissues readily provides a variety of primary cell types that, to varying degrees, can be expanded and further studied/manipulated in cell culture. In addition, PSW-specific molecular markers have been developed that permitted the monitoring of fibroblast, alveolar type II, and vascular endothelial cell types. Reconstitution of 3-D cultures of lung tissues with these cell types is now underway. This novel system may facilitate the development of rare or disease-specific lung tissue models (e.g., to test causes of PSW stranding events and lead to improved treatments for pulmonary hypertension or reperfusion injury in humans). Also, the establishment of a "living" tissue bank biorepository for rare/endangered species could serve multiple purposes as surrogates for freshly isolated samples.
目前，用于研究组织功能和生理学的体外模型，包括对缺氧或环境毒素的反应，都受到限制，并且严重依赖于标准的二维（2-D）培养物，这些培养物使用的是永生的鼠类或人类细胞系。为了开发一种新的、更强大的模型系统，我们一直在寻求建立和扩展原代肺细胞类型以及海洋哺乳动物组织重建的方法。人们对深海潜水侏儒抹香鲸（PSW）的生理学了解甚少，这些知识主要来自于在美国东南部沿海发生的搁浅事件。因此，我们开始开发一种从死亡的搁浅个体中保存活组织和回收活细胞的方法。本报告记录了成功地对 PSW 肺组织进行了冷冻保存。我们从在搁浅事件中几个月前冷冻保存的组织碎片中建立了原代肺细胞类型的体外培养物。冷冻保存的肺组织很容易进行分离，从而提供了各种原代细胞类型，这些细胞类型在不同程度上可以在细胞培养中进行扩展和进一步研究/操作。此外，还开发了 PSW 特异性分子标记物，可用于监测成纤维细胞、肺泡 II 型和血管内皮细胞类型。现在正在进行这些细胞类型的 3-D 肺组织重建培养。这种新系统可能有助于开发罕见或疾病特异性的肺组织模型（例如，测试导致 PSW 搁浅事件的原因，并导致人类肺动脉高压或再灌注损伤的治疗方法得到改善）。此外，建立一个用于稀有/濒危物种的“活体”组织库生物库可以作为新鲜分离样本的替代物，用于多种目的。
