Function/ Services of the CORE:
The function of the Cell Isolation and Tissue Culture Core is to provide efficient, economical, and reproducible cardiopulmonary vascular cell and cardiac fibroblast isolation, characterization, and propagation for investigators using established protocols and new state-of-the-art isolation techniques. In conjunction with isolation services, the Core can provide validation/characterization of isolated cells using flow cytometry; imaging assessments of tissue sections by brightfield and fluorescence imaging; endothelial progenitor cell and microparticle isolation from patient blood.
Mouse endothelial cells can be isolated using Miltenyi gentleMACS™ Dissociator and Miltenyi’s microbead sorting process. We are in the process of developing this new technique to isolate other cell types from this and other species, as well as organs, to be included in our services at a later date.
Cell types can be validated with Miltenyi MACSQuant Analyzer 7 flow cytometer which has 3 color lasers (red, blue, violet) and can analyze 8 colors. This instrument can perform, as a service, phenotypic analysis and functional studies such as cell cycle, apoptosis, cell proliferation, microparticle analysis, quantitation of circulating EPCs. Analysis software is compatible with general flow software such as FlowJo.
Tissue sections can be scanned and analyzed using Aperio CS brightfield 5 slide scanner or Aperio FL fluorescence 5 slide scanner. Analysis software for the CS includes: Area Quantification, Whole Cell Quantification and Microvessel Analysis. FL image analysis software includes: Cytonuclear and Area Quantification applications.