Wake Forest Center for Comparative Medicine Research/Primate Center
The Center for Comparative Medicine Research has been home to the faculty of the Section on Comparative Medicine since the mid-1960s and was a pioneer in use of nonhuman primates in biomedical research. It is one of the largest nonhuman primate research facilities in the US, and Wake Forest University is one of a few universities licensed by the Centers for Disease Control and Prevention to import and quarantine nonhuman primates directly from their countries of origin. The comparative pathology lab has a component within the WFBCCC Tumor Tissue and Pathology Shared Resource. Research training programs (a T32 and a T35) are aimed at pre- and postdoctoral veterinarians. The T32 grant (T32OD010957; PI: J. Mark Cline, D.V.M., Ph.D.) that supports the postdoctoral activity is in its 55th consecutive year of funding. The Center has also made its outreach program a priority, and these extensive activities serve both scientists and the lay community.
The mission of the center is to facilitate the use of animal models, especially nonhuman primates, as patient surrogates to study diseases of human relevance. Ongoing research programs utilize cynomolgus macaques (Macaca fascicularis), rhesus macaques (Macaca mulatta) and vervet/African green monkeys (Chlorocebus aethiops sabaeus, detailed below). These research programs focus on cardiovascular disease and lipid metabolism; effects of aging, obesity and diabetes; substance abuse – both cocaine addiction and alcohol abuse; hypertension and vascular disease; breast, uterine and prostate cancer; neuroscience and behavior; bioterrorism studies – vaccine development and radiation countermeasure studies; women’s health; and nutritional influences on human disease processes.
The Primate Center is located on the 200-acre Clarkson Campus, located nine miles south of the Medical Center/Hawthorne Campus. This complex includes 32 buildings, 120 employees, and about 1,000 nonhuman primates. This campus has 15 housing buildings for nonhuman primates, along with animal housing for sheep, animal support areas, totaling more than 80,000 net square feet of animal housing space, not including pastures. Additionally, the Primate Center provides space for faculty offices and research laboratories, support staff, and computer and data services staff. A centralized data room and conference rooms are available. Information on WFPC primate research colonies, facilities, and personnel can be found at http://www.wakehealth.edu/wfupc/.
Vervet Research Colony
This NIH-supported national resource (P40 OD010965; PI: Dr. Kaplan) consists of approximately 450 vervet or African green monkeys (Chlorocebus aethiops) living in 16 breeding groups. The Vervet Research Colony (VRC) was transferred from UCLA to the WFSM in 2007. It was initially founded in 1975 with 14 vervets captured from St. Kitts and the West Indies. Between 1976 and 1985, 43 wild caught animals were added. The current population of the VRC includes more than 450 descendants from the 57 original founders (29 females, 28 males), with 24 of the original matrilines now in their 3rd to 7th generation. Colony management practices reflect the natural social composition of vervet monkey groups in the wild. Housing consists of 16 enclosures with 1000 sq. ft. of outdoor and 300 sq. ft. of indoor space, each containing one breeding group. Infants and juveniles remain in the natal group with their mothers and female kin. Males are removed at adolescence and transferred to other groups, and adult males are rotated between groups at 3-4 year intervals, to mimic the natural processes of emigration and immigration. The colony is managed to maintain genetic variability, avoid inbreeding depression, and promote long-term viability of the population.
Research Facilities
Surgery Facilities (Melaney K. Gee, DVM, Director) - The surgical suite at the WFPC consists of rooms for surgeons and instrument preparation, animal preparation and recovery, and two operating rooms. Trained and experienced animal health technicians are available to assist veterinarians and research investigators with surgical procedures. This surgery suite is equipped with gas anesthesia machines with isoflurane vaporizers, water-heated thermal blankets and forced–air warming blankets (Bair Hugger), pulse oximetry, electrocautery, and a Vet specs 9062 V vital signs monitor. On average, 350 procedures per year are performed in this suite.
Animal Radiology Suite (Melaney K. Gee, DVM, Director) - The Animal Radiology Laboratory is located in Room 120 of Building 08. This facility is equipped with an Innovet Select Veterinary Radiographic System (40-125 kVp, 1-300 mA), and an automatic film processor model AFP14XL. The room also has a built-in film cabinet and personal protective equipment. The equipment is utilized for both clinical/diagnostic and experimental procedures. Our annual activity averages approximately 100 procedures.
Hematology Laboratory (Multi-user laboratory) - The Hematology laboratory supports the research of all center investigators and contains all supplies and equipment necessary for processing blood and CSF samples including multiple ultracentrifuges.
Primate Imaging Laboratory (Multi-user laboratory) - The facility is equipped with a Hologic Discovery A dual X-Ray Bone Densitometer, a Hologic (vendor built) PC with a 75 GB hard drive, Acer Monitor, Hewlett Packard Officejet Pro K5400, one quality control phantom (Spine 2928), general anesthesia (isoflurane) machine and recovery area. The equipment is utilized mainly for experimental procedures to measure whole body, anterior-posterior spine, and compartmentalized (appendages) bone density. Whole body and abdominal composition (% fat) can also be measured. The coefficient of variation has been determined for the following measurements (whole body BMC -1.2 ± 0.3%, lumbar spine BMC - 1.8 ± 0.6%, and lumbar BMD 1.5 ± 0.5%. The equipment will be used to measure body composition.
Primate Nutrition Laboratory (Susan Appt, DVM, Director) - This lab (352 sq. ft.) is responsible for the preparation of all experimental diets for nonhuman primates. There is an adjacent temperature-controlled storeroom for diet ingredients (660 sq. ft.) and a large room (1,188 sq. ft.) with about 25 freezers for storage of prepared diets. This laboratory has been providing specialized diets for our animal studies for over 40 years and currently prepares approximately 60 different diets per for ongoing investigations involving monkeys, rodents, and rabbits. Diets are prepared based on specific formulations developed in consultation with the laboratory supervisor, color coded, and then stored in study-specific freezers. Furthermore, samples for chemical analysis are taken from every diet lot. This laboratory allows investigators to mimic closely the environmental context within which human disease develops.
Comparative Behavior Laboratory (Jay R. Kaplan, PhD, Director) - This laboratory consists of two rooms. The first room (600 sq ft) houses computers, data records, EKG telemetry equipment, and space for sample labeling, preparation and data storage. The second room (120 sq ft) serves as office space for two laboratory technicians. Dr. Kaplan has additional space (150 sq ft) for his lab supervisor, computer storage and data archival. The staff of this laboratory is tasked with evaluating the behavior and social status of group-housed monkeys and their relation to disease susceptibility and resistance.
Reproductive Medicine Laboratory (Susan E. Appt, DVM, Director) - This laboratory consists of approximately 425 sq.ft. This 425 sq. ft. laboratory is shared with Drs. Clarkson and Kaplan and is focused on studies of ovarian structure and function. The investigators in this laboratory have special expertise in the evaluation of ovarian dysfunction in relation to genetic and environmental factors and the development of a nonhuman primate model of the menopause.
Aging Physiology Laboratory (Kylie Kavanagh, VMS, MS, MPH, Director) - This is a 600 sq. ft. laboratory located in the Janeway Research Building in the downtown Innovation Quarter. There is a fume hood, sinks, vacuum, refrigerator and –20ºC and –80ºC freezers for use. General laboratory equipment is available (pipettes, centrifuges, electrophoresis apparatus, vortexes, etc). Desk space and computers are available in the laboratory area for both technicians and students to work. Major equipment relevant to most research applications are available either within Kavanagh’s lab or the shared resources of Lipid Sciences laboratories. The Lipid Sciences Program has 500 square feet of shared equipment space, 200 square feet of shared cold room space. The Program includes a GC/Mass Spec core laboratory, a lipid and lipoprotein core laboratory (for fatty acid and sterol measurements by gas-liquid chromatography, ELISA assays for apolipoprotein quantification, and FPLC separation of plasma lipoproteins), a dark room, an autoclave and a conference room with video conferencing capabilities. Shared equipment in this area includes a dark room, two tissue culture rooms (415 sq ft each), four cold rooms, -80°C. freezer room, ABI 7000 real time PCR, FujiFilm Luminescent Image Analyzer with cold CCD camera for Western blots and Phosphorimager (LAS-3000), HP gas-liquid chromatograph, Genios microplate reader (for fluorescence and absorbance measurements), AlphaInotech Image Analyzer, Beckman gamma counter, Beckman liquid scintillation counter, BD FACSCalibur (4 channel) flow cytometer and FloJo software, and Olympus microscope equipped with digital camera and Image Pro Plus software. Dr. Kavanagh also has an office at the WFPC.
Clinical Chemistry and Endocrinology Laboratory (Thomas Register, PhD, Director) - The Clinical Chemistry and Endocrinology Laboratory is located in Building 12 and has approximately 650 sq.ft. of space for clinical chemistry determinations, hematologic procedures, and microbiologic and parasitologic procedures. A senior-level technician (Ms. Maryanne Post) with more than 25 years' experience supervises the laboratory. Additional equipment includes an Ace Alera Chemistry System and a HEMAVET 950 FS multi-species hematology system, allowing processing of more samples. Appropriate centrifuges, Coulter counters, and a Beckman DupR 540 spectrophotometer are available. The facility is equipped with an Antek Labdaq Laboratory Information System (LIS), Tecan GENios Microplate Reader (with Magellan Software) capable of measuring absorbance, fluorescence and luminescence, a Tecan Columbus Microplate Washer, and a Packard 9000 Gamma Counter for radioimmunoassay procedures.
Molecular Biology Laboratory (Thomas Register, PhD, Director) - Research and core laboratories are equipped for cell and tissue culture, protein, RNA, and DNA electrophoresis and other analyses, including ELISA, protein immunoblotting, nucleic acid blotting (Northern, Southern), RT-PCR and quantitative real time PCR, as well as histology, morphometry, immunohistochemistry and in situ hybridization. Major equipment includes a Molecular Dynamics STORMR 860 Imager and Dell Optiplex GX1 computer equipped with ImageQuant 5.1 Software, Perkin Elmer 9600 Automated Thermal Cycler, ABI Prism 7000 and 7500 FAST Sequence Detection Systems for real time quantitative RT-PCR, Hybaid Mini Hybridization MKII Oven, Perkin-Elmer HPLC System with UV-Vis, fluorescence, and electrochemical detectors, microtiter plate readers and microplate washers, a CCD camera attached to an Analytical Imaging Station (AISTM) from Imaging Research Inc. (Brock University, Ontario, Canada), and HP Color Laserjet, 4500HP, and HP Photosmart printers. Other standard equipment includes water-jacketed thermostated CO2 incubators, a laminar flow safety hood, high-speed refrigerated centrifuges and ultracentrifuges, liquid scintillation and gamma counters, a Reichert-Jung Cryocut cryotome, low-temperature freezers, liquid nitrogen dewars, Speed Vac, and fluorescence and UV-visible spectrophotometers.
Behavioral Neurobiology Laboratory (Carol Shively, PhD, Director) - This 800 sq ft. laboratory consists of three rooms. The laboratory provides space for several technicians and graduate students. The lab is equipped with an ECG Telemetry System, an Actigraphy System, PC’s and laptop computers for behavioral data collection and analysis. Included is storage space for clinical supplies, fixed tissues, and several -70°C freezers for frozen tissues.
Diabetes and Lipid Laboratories (Kylie Kavanagh, VMS, MS, MPH, Director) - These laboratories include approximately 1,000 square feet and include facilities for studies on diabetes, inflammation/oxidation, and lipid/lipoprotein studies.
Comparative Pathology and Experimental Atherosclerosis Laboratories (Director, Dr. Register) - These laboratories are designed and equipped for processing fixed and frozen tissues for determining the extent, severity, and histologic characteristics of atherosclerotic lesions, including histochemical and immunochemical staining and for computer-based histomorphometric imaging. Additionally, inflammatory marker analyses are performed in these laboratories.
Comparative Pathology Research Laboratory (J. Mark Cline, DVM, PhD, Director) - This clinical studies/image analysis lab is a 500 sq. ft. laboratory equipped with desks and networked computers for 4 technicians, 2 light/fluorescence microscopes connected to computer imaging workstations, hoods and wet lab space. This lab serves as a staging area for primate clinical studies and includes equipment for in vivo studies (two Sonosite 5-8 mHz and one 13 mHz portable ultrasound machines, anesthesia machines, surgical supplies, pulse oximeters, and Heska hematology, clinical chemistry analyzers, and blood gas analyzers).
Histology and Immunohistochemistry Laboratories (Dr. Cline, Director) - This laboratory is supervised by Dr. Cline in support of all researchers at the WFPC. The Histology Laboratory has approximately 375 sq. ft. which includes a separate room for tissue processing, two tissue processors and an embedding station for paraffin embedding, 3 built-in hazardous-materials hoods, and two networked PCs. The laboratory serves as the resource for investigators requiring histologic slides, and has expertise in paraffin, plastic and frozen sections, and a wide variety of special stains. The laboratory has particular expertise in the preparation of microscopic slides for quantitative morphometric evaluation, especially vascular and reproductive tissue. In addition to production of histologic and microscopic slides,
Glass and Paraffin block archives (Multi-user) - Staff in this laboratory maintains the glass slide and paraffin block archives, which are cross-referenced for rapid retrieval of both diagnostic and experimental material.
Adjacent to this lab is the Immunohistochemistry Laboratory, a 500 sq. ft. laboratory fully equipped for immunohistochemistry and in situ hybridization studies. It contains a Reichert-Jung Cryocut cryotome, 2 microtomes, Mettler balances, an Eppendorf centrifuge, a refrigerator, microwave and conventional ovens, a fume hood. Computer resources include 2 networked workstations, one of which is Staining and quantitative assessment is routinely done for estrogen and progesterone receptors, cell cycle markers (p21, p27, Ki67, cyclin B), apoptosis (cleaved caspase 3, TUNEL), inflammatory markers (C-reactive protein) and a variety of other markers.
Also in the same area are 1) a 100 square foot microscopy room housing a 7-headed teaching microscope and a digital photomicroscopy unit, 2) a 50 square foot storage room for histoslides, and 3) a 75 square foot Image Analysis laboratory, equipped with two networked computer workstations, configured for histomorphometry and quantification of immunohistochemical stains by connection to video-microscopy units consisting of a Nikon Labophot II microscope with epifluorescence capability, Sony or Luminera color video cameras, and software (Image-Pro Plus) for color image analysis.