CORE--TISSUE CULTURE

此项目1-4需要培养内皮细胞(ECs)和/或分离的中性粒细胞 计划项目。核心B将主要服务于罗切斯特项目(2、3和4)的要求，尽管核心和项目1之间将有一些正在进行的双向技术转让。我们发现，通过结合对内皮细胞培养系统的许多常规技术支持，我们能够显著节省成本和精力。然而，这一核心设施的主要原理在于我们从目前项目期的经验中获得的经验，即对于内皮细胞，在不同的生长形态-微通道、微载玻片、平行板系统等-中对内皮细胞的生长有严格且不同的要求。例如，成功地将HUVEC生长到微载玻片中汇合的程序是复杂的，并且在种子接种方案、预期汇合时间以及对在微载体珠上生长的细胞的维护要求方面有所不同。这些 配置需要更多的时间来设置和维护，并且更容易受到小的程序事故的影响。我们预计技术开发将至少在下一个资助期的早期进行，例如确定最符合项目3中微吸管研究需要的微载体珠子的成分和涂层。随着程序的建立，它们将用于计划的研究；到那时，核心技术人员将投入相对更多的精力来生产在适当配置中生长的足够细胞，用于各种项目计划的研究。因此，通过使用共同的设施来继续工作的这些技术方面，同时 根据不同配置的需要，监督合流电池系统的生产，我们期望更有效地支持个别项目的需求。这确实是我们在过去几年的经验。

Cultured endothelial cells (ECs), and/or isolated neutrophils are required by Projects 1 - 4 of this program project. Core B will primarily serve the requirements of the Rochester projects (2, 3, and 4), although there will be some ongoing technology transfer in both directions between the core and Project 1. We have found that by combining much of the routine technical support for the endothelial cell culture systems we are able to effect a significant saving in cost and effort. However, the major rationale for this core facility resides in our experience from the present project period that, for endothelial cells, there are stringent and differing requirements for growing endothelial cells in the various growth configurations -microchannels, microslides, parallel plate systems etc. For example, procedures to successfully grow HUVECs to confluence in the microslides are complex, and different with respect to seeding protocols, expected time to confluence, and maintenance requirements for cells grown on microcarrier beads. These configurations take more time to set up, and to maintain in culture, and are more easily contaminated by minor procedural accidents. We expect that technical development will be ongoing during at least the early part of the next grant period, for example in determining composition and coatings on microcarrier beads that best meet the needs of the micropipette studies in Project 3. As procedures become established, they will be used in the planned studies; at that point, relatively more of the effort of the core technician will be directed to producing sufficient cells grown in the appropriate configurations for studies planned by the various projects. Thus by using a common facility to continue these technical aspects of the work while overseeing the production of confluent cell systems as needed in the various configurations, we expect to more efficiently support the needs of individual projects. This has indeed been our experience during the last several years.