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.
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