CORE--TISSUE CULTURE

本项目1 - 4需要培养的内皮细胞（EC）和/或分离的中性粒细胞。 程序项目。核心B将主要满足罗切斯特项目（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。我们已经发现，通过结合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，该核心设施的主要原理在于我们从当前项目期间获得的经验，即对于内皮细胞，在各种生长配置-微通道、微载玻片、平行板系统等中生长内皮细胞有严格且不同的要求。例如，在微载玻片中成功生长HUVEC至汇合的程序是复杂的，并且在接种方案方面不同，预期的汇合时间和在微载体珠上生长的细胞的维持要求。这些 配置需要更多的时间来设置和在培养物中保持，并且更容易被轻微的程序事故污染。我们预计，至少在下一个资助期的早期，技术开发将继续进行，例如确定最能满足项目3中微量移液管研究需求的微载体珠的组成和涂层。随着程序的建立，它们将被用于计划的研究中;在这一点上，核心技术人员的相对更多的努力将被用于生产足够的细胞，这些细胞在适当的配置中生长，用于各种项目计划的研究。因此，通过使用一个共同的设施继续这些技术方面的工作， 通过监督各种配置中所需的汇合细胞系统的生产，我们希望更有效地支持各个项目的需求。这确实是我们过去几年的经验。

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.