The primary justification for the requested system is the current lack of flow cytometry and cell sorting at the Milwaukee VA despite a high demand for such procedures. The requested system is a BD FACSARIA Fusion Special Order flow cytometry and cell sorting system with all components necessary to execute multicolor flow cytometry and cell sorting experiments. The instrument will consist of five lasers, which allows for detection of 18 fluorescent parameters. This will enable researchers at the Milwaukee VA to detect a large variety of cellular markers in the same cell. This instrument is produced by the market leader BD. VA researchers have previously used flow cytometry and cell sorting at the core facilities of the Medical College of Wisconsin. The same instruments and software are represented at the core facilities, which would allow researchers at the VA to continue their experiments without technical adjustment. The research projects which would strongly benefit from the presence of a flow cytometer and cell sorter at the VA include a variety of conditions relevant to both Veterans and the general population. Dr. Kurpad is using diffusion tensor imaging (DTI) analysis of the injured spinal cord at the lesion site and proximal to the lesion. Flow cytometry will be used to verify imaging results and to identify and correlate cell activation, cell death and cytotoxicity to the DTI results. Dr. Jacobs is investigating ischemia-reperfusion lung injuries, a commonly encountered condition after lung transplantation. Flow cytometry will be performed on cells derived from bronchoalveolar lavage and on enzymatically digested lung tissue and will provide information about cell signaling and mitochondrial properties. Furthermore, the result of therapeutic interventions will be assessed by flow cytometry. Dr. Rosenthal's research is focused on a form of degenerative arthritis, the calcium pyrophosphate deposition disease. The cell sorter would be used to sort populations of successfully transfected chondrocytes which can then be used for further experiments. Dr. See's project addresses the induction of oxidative stress in urothelial carcinoma cells by BCG bacteria. This oxidative stress is a critical component for the antitumor activity of BCG. Flow cytometry will be used to detect treatment effects in terms of cytotoxicity and cell death. Dr. Kroner-Milsch is investigating the role of inflammation after spinal cord injury (SCI).
She aims to reduce secondary damage by specifically blocking cytokines and chemokines which are upregulated following SCI. Cells derived from injured tissue will be analyzed for production of cytokines and reactive oxygen species. Dr. Glavaski-Joksimovic's research focus is the neuroprotective effect of adult mesenchymal stem cells (MSC) after traumatic brain injury (TBI). Flow cytometry and cell sorting will be used to study and sort different MSC subpopulations and compare their effects in TBI models. Dr. Beilke is investigating the role of HTLV-2 virus as a modulator of immune response. Tax2, a HTLV-2 protein, is a strong chemokine regulator and plays a role in anti-viral activity against HIV-1. Flow cytometry is frequently applied in his studies to detect expression of surface molecules in specific cell populations. Dr. Budde's research focus is the development of noninvasive MRI biomarkers for SCI and TBI. A flow cytometry and cell sorting instrument will be necessary for cell based therapy studies with a high promise of neuroprotective and regenerative effects. Dr. Ridolfi investigates the role of 5-HT3 and 5-HT4 receptors after rectal cancer resection. Flow cytometry will be used to assess expression density in his rat model of parasympathetic damage. In summary, access to a flow cytometer and cell sorter at the Milwaukee VA would accelerate research projects currently done at the VA and open new research opportunities for VA investigators.
Researchers at the Milwaukee VA work on various conditions that are significant public health concerns. The requested instrument, a flow cytometer and cell sorter, is a tool that enables researchers to analyze cells derived from different tissues with regard to their characteristics, production of proteins or toxic factors, cell cycle and signal transduction. Furthermore, it allows for specific sorting and collection of labeled cells in order to further analyze these cells, take them into culture or transfer them into animals as a treatment. Currently, no such instrument is available at the Milwaukee VA. Researchers use comparable instruments at MCW, but the facilities are heavily used and transport process and time have a negative impact on cell survival, reliability of stainings and sterile conditions of cells. The availability of this instrument on site would accelerate research and improve the quality and scope of conducted experiments. The goal is to clinically translate information gained from these studies, which will ultimately help to contribute to better health care for Veterans and the general population.
