1998-- https://taskbook.nasaprs/peer_review/taskbook/micro/mg98/agreement.html In 1998 new studies have been launched at the Center to research endometriosis using RWV Bioreactor to culture endometrium biopsies, Lyme disease using innoculation of RWV- cultured human tissue, diabetes by maintaining human islet cells in the RWV, and arthritis by culturing synovial tissue from rheumatoid arthritis patients. Study of HIV mucosal transmission in RWV-cultured tissue was recently started in collaboration with French scientists. -- II. National Cancer Institute A. Long term maintenance of human prostate tissue in the RWV Bioreactor. B. Study of Lyme disease in RWV Bioreactor 8. Assessment of the RWV as a universal pathogen culture system Primary Investigators: Dr. Steven Hatfill, Laboratory of Cellular and Molecular Biophysics, NICHD, NIH and USAMRIID, Dr. Paul Duray, Department of Pathology, NCI, NIH, and Dr. Michael Bray, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Ft. Detrick, MD. Aim of Experiments: To demonstrate the potential ability of the RWV Bioreactor to act as a universal pathogen culture system for the primary isolation of previously unrecognized pathogens during outbreaks of emerging disease. By demonstrating the ability of the RWV to culture a variety of known pathogens of different classification, i.e. RNA viruses, retroviruses, DNA viruses, parasites, spirochetes, etc., we hope to demonstrate the applicability of the RWV system for de novo primary pathogen isolation protocols. BACKGROUND: At present, the de novo isolation of previously unknown or unrecognized emerging disease pathogens requires elucidating the proper culture media or permissive cell line applicable for growth of the pathogenic agent in culture. A few examples serve to illustrate the difficulty inherent in this process using conventional technology. The 1970s outbreak of Legionaries disease required months to identify the causative agent as a bacterium. The HIV-1 retrovirus required over 2 years to isolate and in the 1993 outbreak of Myuro Canyon disease in the four-corners region of the United States it took 7 weeks to grow the Hantavirus in culture. Clearly these timelines are unacceptable with respect to many public health threat scenarios for emerging disease agents. By utilizing the ability of the RWV to maintain a normal three-dimensional cytoarchitecture and microenviroment for a number of tissues, the possibility of using human tissue explants for primary pathogen isolation, becomes a distinct possibility. Experiment Design: Attempts will be made to culture a variety of infectious pathogen in the same simple medium (RPMI-1640, 15% FCS) containing human tonsil tissue explants maintained in the RWV Bioreactor. In addition, a human liver and epithelial tissue equivalent will be formulated from established cells lines grown on Cytodex 3 microcarrier beads. These will be co-cultured with the tonsil explants. Known pathogens which have proved to be difficult to isolate by normal protocols, will be introduced into the RWV culture and allowed to incubate with the tissue and tissue equivalents for two weeks. Continuing Results: 1. It has been previously demonstrated that the spirochetal organism Borrelia burgdorferi, the agent of Lyme disease, grows and proliferates in this system. The system has been shown to support productive infection with HIV-1 strains. Ebola virus was also shown to the amplified in this culture system. more.......
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