Resources and environment – Institute for Regenerative Cures, Sacramento CA
The UC Davis Stem Cell Program that Dr. Nolta directs is housed in the 109,000 SF UC Davis/CIRM Institute for
Regenerative Cures (IRC, Figure below). The IRC was funded through a $20 million dollar grant from the California
Institute for Regenerative Medicine (CIRM), an agency which has funded over $50 million total for members of the
UC Davis Stem Cell Program. UC Davis has a strong track record in translational biomedical science. Building on
the scientific assets and expertise of one of the largest life science faculties in the nation, the UCD Stem Cell
Program has provided a visible intellectual home and a potent coordinating force for 145 faculty investigators who
are collaborating on regenerative medicine research, organized into more than 15 disease teams: Peripheral vascular
disease, heart disease, organ bioengineering and
regeneration for liver, lung, bladder and kidney,
blood disorders including leukemia and sickle
cell anemia, Immune system disorders including
HIV, restoration of hearing and sight, bone
repair and cartilage regeneration, studying
cancer stem cells to prevent relapse, skin repair
including burns and non-healing ulcers,
neurodevelopmental disorders including fragile
X and autism spectrum related disorders, and
neurodegenerative
diseases
including
Huntington’s disease, Alzheimer’s disease,
Parkinson’s disease, and ALS or Lou Gehrig’s
disease. Each of these teams is comprised of
leading researchers and clinicians who are
exploring the most promising approaches that
can lead to the greatest advances in improving
for Regenerative Cures. The state-of-the-art 109,000
patient health. These groups are supported by an Institute
2
NIH-funded Clinical Translational Science ft Facility enhances collaborations among physicians,
Center (CTSC) and a spectrum of institutional researchers and collaborators, and provides a Good
resources such as access to a wide range of Manufacturing Practice (GMP) Facility for Cellular Therapies
animal models including outstanding mouse and other stem cell-based therapeutics.
facilities, an immune deficient mouse core and
an NCRR-funded non-human primate facility, sophisticated analytical and imaging technologies and expertise, a 6
suite, state of the art Good Manufacturing Practice facility, and an integrated academic/clinical services health
system structure that is poised to effectively move promising innovations in the field of regenerative medicine into
clinical trials. This interdisciplinary research initiative brings together faculty at the School of Medicine with other
research groups and investigators to maximize synergy in moving regenerative therapies into the clinic.
The Institute for Regenerative Cures includes a state-of-the-art 6-suite Good Manufacturing Practice (GMP) facility
that will enable researchers to readily move cellular therapy development candidates that have previously been
studied in animal models into clinical trials in human patients. The building serves as a hub for basic, translational,
and clinical regenerative cures research at UC Davis. Numerous collaborations are currently in progress throughout
the Sacramento and Davis campus. The center serves as a centralized site for bringing together researchers in stem
cell-mediated repairs of different tissues and organs, to share equipment and ideas.
Translation to Clinic: The Good Manufacturing Practice (GMP) facility and CTSC will allow direct translation of
the good laboratory practice (GLP) studies done in Dr. Nolta’s laboratory into clinical trials, if they are successful.
Dr. Nolta is the Scientific Director of the 6-suite GMP facility in the UC Davis Institute for Regenerative Cures, and
has had 15 years of experience in implementing and conducting gene and cellular therapy trials at the GMP level.
We have an impressive team of translational researchers cross-trained in GMP manufacturing, and administrative
and statistical oversight and project management for filing of investigational new drug applications to the FDA,
through our stem cell program and the clinical translational science center, to help move promising research forward
from the bench to the bedside.
GMP Resources: The GMP facility located in the Institute for Regenerative Cures at the University of California
Davis, School of Medicine, is a multi-use, 6 manufacturing suite, Class 10,000 facility, for manufacturing of
clinically applicable products under current Good Tissue Practice, current Good Manufacturing Practice and USP
797. All 6 manufacturing rooms are independent of each other, applying one way personnel, product and waste flow.
6 different products can be manufactured without interfering with each other. The manufacturing, entry and exit
areas are separated by air pressure gradients. An elaborate interlock system controls opening and closing of access
doors, therefore avoiding cross-contamination of products by air turbulences. The GMP facility features an
electronically controlled and automatically monitored environment, with 7 day per week manual monitoring as well.
A strict QC/QA program with environmental cleaning and monitoring procedures is implemented to assure strict
quality control of facility and products, and freedom from microbiological contamination. In a type C meeting, the
plans for the facility were discussed with and approved by the FDA. This facility provides a state of the art
environment for the manufacturing of novel therapeutics for human clinical applications. The trials for our 15 UC
Davis Disease Teams and 5 collaborating Disease Teams will be performed there.
Translational resources/ Good Laboratory Practices: Prior to the actual expansion of cells for trials, the
technique validations and in vivo testing can be done adequately at the level of Good Laboratory Practice, in a nonGMP setting. We are experienced in these techniques and in the extensive record-keeping and source document
retention required to validate cells and reagents for clinical applications. The 5-laser Cytopeia Influx Flow Sorter,
currently in our Stem Cell Program’s sorting facility, is encased in a biosafety cabinet for GMP-grade sorting. The
CliniMACS immunomagnetic isolation device, Sysmex and FACScalibur for the clinical trials are in the same
designated scale-up area in the building that temporarily houses the Stem Cell Program. The scale-up and validations
are ongoing there.
For the in vivo safety trials, the principal investigator has three separate rooms within a contiguous suite, dedicated
solely to the breeding and care of her novel strains of immune deficient mice, in the Research III Barrier Facility on
the UCD Medical Center Campus, one block from the lab. There is a large laminar flow hood in each room. Two
technicians care for the mice, monitoring each cage twice daily, preparing antibiotic-treated water and irradiated
food, and changing microisolator cages twice a week. The health of the animals is checked daily. Sublethal
conditioning is done by irradiation, using the animal irradiator just purchased for the Stem Cell Program, and located
in that facility, across the hall from the animal suite. This facility is run as a core, on a cost recovery basis, and will
be moved into the large vivarium in the new Institute for Regenerative Cures in late 2010.
The UC Davis stem cell program currently has 145 faculty members, including basic and translational scientists and
clinicians, assembled into 15 disease teams. Each team is working toward the use of stem cell therapies in clinical
trials. Systems in place in the Stem Cell Program and in the NIH-funded CTSC will help many of our disease teams,
and those of our collaborators, to safely and effectively bring their cures to FDA-approved clinical trials to help
those in our community, and throughout the United States, who so desperately need them.
CORE SERVICES in the Institute for Regenerative Cures
Stem Cell Program Core Facilities and Resources: The Stem Cell Program provides multiple cores and services
summarized in the figure below, including a retroviral and lentiviral vector core, a Flow sorting and FACS analysis
core, a microscopy core, an immune deficient mouse core with tissue damage models already in place, and provides
scale-up services for clinical cellular therapy trials to be done in the GMP facility.
The Flow-Assisted Cell Sorting (FACS) Core for the Stem Cell Program is run by Carol Oxford, Technical
Director, and her staff. Ms. Oxford, a well-respected expert in her field, has overseen the Optical Biology Shared
Resource at UC Davis for the past decade. She operates the cell sorter, asists with experimental design, maintains all
instruments and is in charge of scheduling their use and reporting usage for billing. The Cytopeia InFlux is a high-
speed cell sorter with six lasers and 16 parameter measurement capability, and was purchased for the Stem Cell
Program. This will be the main sorter in the IRC, and can detect 14 simultaneous fluorescent parameters. Up to 4
populations can be sorted simultaneously. A biohazard containment system is in place. At the time of its last
upgrade, this instrument was one of only three in the world with this capability. The FC500, new NIH ARRAfunded LSRII, and other cytometers complete this core and can be used by any PI, technician or trianee in the
program, under Ms. McLaughlin’s training and guidance.
Microscopy and videomicroscopy Core. BioStation IM (time-lapse): The BioStation IM is an invaluable tool to do
long term imaging of live cell cultures by both phase and epifluorescence microscopy. Its compact design includes
an incubator, microscope, and a cooled CCD all housed in a single system that allows for easy to use observation of
cell cultures in a stable, sterile environment. Eclipse Ti-U (inverted): The Eclipse Ti-U is a versatile inverted
microscope equipped with the ability to perform phase, differential interference contrast, and epifluorescence
imaging on live cultures in a variety of tissue culture formats. It is a platform for monitoring, imaging, and
performing short-term high speed imaging of live cell cultures by multiple modalities and employing a perfect focus
system to ensure sharp images. The Ti is also extremely upgradeable, allowing us the potential to expand the system
to use deconvolution or confocal imaging techniques, automated imaging and screening, long-term time lapse, or
many other functions that would normally require a separate microscope. This system is used heavily in to monitor,
image, and document cell cultures.
The Stem Cell Program’s Vector Core provides a centralized service for the development and production of viral
vectors necessary for gene transfer in research experiments and pre-clinical studies. Expert technical staff will
consult with investigators to plan and develop vectors to fit individual project requirements, design and construct
novel vectors and generate purified recombinant moloney murine retrovirus and lentivirus. Quality control testing
will include vector titering and replication-competent retrovirus (RCR) and Lentivirus (RCL) assays. The core has
been providing novel non-integrating lentiviral vectors to generate iPSC from patient samples for the majority of the
groups working in the stem cell program. The UC Davis Good Manufacturing Practice facility in the Institute for
Regenerative Cures provides additional services to produce validated high titer vector supernatant for human clinical
trials, for UC Davis investigators and outside parties under contract.
Specific Pathogen Free Barrier Facility (SPF) Vivarium: The facility consists of approximately 10,5000 asf of
disease-free housing for experimental animals comprised of twelve holding/ procedure rooms, and two that can be
configured as holding or procedure rooms. These rooms are separated into four suites of three rooms and one suite
of two rooms. The top 30 users of animals at UC Davis mimic the national average of 248 cages per investigator.
Using this average of 248 cages per investigator as the standard, this initial configuration will support 15 of these
national average senior investigators. The following resources are situated within the animal facility: A tissue culture
room for cell isolation, the animal irradiator for transplantation procedures, and four separate imaging/behavioral
assessment rooms. The potential modalities include micro Computed Tomography (CT), micro Positron Emission
Tomography (PET) ultrasound, optical/fluorescence IVIS, x-ray with fluorescent overlay and other imaging
equipment.
Immune deficient mouse core for analysis of human stem and progenitor cells in tissue repair models: Dr.
Nolta has established the Immune Deficient Mouse Core at the UC Davis Sacramento campus, operating in
temporary vivarium space until the new Institute for Regenerative Cures Vivarium was ready. The facility is cared
for by the outstanding Center for Laboratory Animal Science (CLAS) and veterinary staff at UC Davis. The core is
run through the Stem Cell Program by Jeannine McGee. The core currently performs experimentation in
NOD/SCID, NOD/SCID/B2M null, and NOD/SCID/ IL2Rg (gamma chain)-/- mice obtained from Jackson
Laboratories, for IRC faculty and outside contractors. Xenotransplantation models are the Nolta lab specialty, and
Dr. Nolta has worked with immune deficient mice for over two decades. In addition to the commercially available
strains listed above, Dr. Nolta supplies the NOD/SCID/MPSVII immune deficient mouse model, which allows the
contribution from transplanted human stem cells to a damaged tissue to be rapidly visualized without upfront
marking or reliance on cell surface markers. This is important because we have demonstrated that stem cell surface
phenotype is dramatically altered during migration into sites of tissue damage and inflammation. This model also
allows FACS- based recovery of cells that have homed into the region of tissue damage, since we have a membranediffusible substrate for GUSB that is cleaved to a fluorescent moiety only in cells with high enzyme levels. This is
allowing us to recover human stem cells that have successfully homed into sites of damage in different tissues, and
to compare them to the transplanted population and to cells that lodged non-specifically rather than homing.
Comparisons are done through the outstanding proteomics core in the UCD Genomics building.
The Clinical Translational Science Center (CTSC), housed in 16,000 sq. ft. designated space in the same
warehouse occupied by the Institute for Regenerative Cures, links key clinical and translational research and training
initiatives, provides increased access to important technologies developed by our scientists, and links these
technologies to our clinician partners in the UC Davis primary care network (PCN), our Community Hospital
Network and TeleHealth Program. The CTSC integrates previously isolated functions, (biostatistics, informatics,
regulatory and IRB functions, clinical trials, and grants and contracts) into a single administrative home, adjacent to
the Stem Cell Center currently under development. It is a specific goal for the UC Davis CTSC to create a
recognized, highly visible, academic home for clinical and translational research benefiting the diverse communities
and populations that UCD serves in inland Central and Northern California, a geographically large region with a
rapidly growing population. The CTSC also streamlines, consolidates, and amplifies services supporting clinical and
translational research and education to enhance their impact and optimize utilization, and promotes professional
interest, among trainees and established scientists, in conducting collaborative translational research.
Telemedicine: We have used tele-technology to connect the UC Davis Health System (UCDHS) to our community
partners not only in Sacramento, where UCDHS is based, but also throughout inland Central and Northern
California. This UCDHS-supported, nationally prominent network, housed in the Center for Health and Technology,
underscores the determination of UC Davis to share our discoveries of new science that improves human health with
our larger constituencies. Regenerative medicine classes and lectures are being shared throughout this network and
therefore our developments are of benefit to multiple centers. Trainees at UC Merced routinely join Regenerative
Medicine lectures at UCD through Tele-technology. The UC Davis CHT is one of the broadest and most innovative
TeleHealth programs in the world. The cybercafé and networking space in the CRCCD provide a teleportal to other
academic sites and rural communities to teach regenerative medicine to physicians, trainees and community
members.
RESOURCES
FACILITIES: Specify the facilities to be used for the conduct of the proposed research. Indicate the
performance sites and describe capacities, pertinent capabilities, relative proximity, and extent of
availability to the project.
Laboratory: Dr. Nolta directs the large shared translational “bench to the bedside” laboratory which has over 8000
SF of space in our new 109,000 SF UC Davis Institute for Regenerative Cures. The institute contains a GMP facility
and mouse barrier facility, as well as bench space for approximately 15-20 PIs. Among the other PIs in our institute
are those working in fifteen different disease teams. These PIs and their teams work in the large 12,000 assignable
square foot shared stem cell discovery laboratory directly across the hall. Investigators have ample opportunity to
interact with those investigators through our weekly large and small group meetings and daily interactions in the
large common lab spaces, meeting and coffee areas.
Major Equipment: Major equipment in Dr. Nolta’s shared translational laboratory includes centrifuges, inverted
phase microscopes, videomicroscopy chambers and equipment, Karyotyping equipment, waterbaths, PCR machines,
fluorescent microscope, ELISA plate reader, CO2 tissue culture incubators, high-powered dissection microscopes,
microcentrifuges, Refrigerators; Freezers (-20°C); (-70°C); large and small Western blot apparatus, power supplies
and transfer equipment, 3D gel apparatus and power supply; gelboxes and power supplies for nucleic acids, gel dryer
and hydro-tech vacuum pump, ultracentrifuges, preparative centrifuges, lyophilizers, speed vac, autoclave, and 24
LN2 tanks in our stem cell bank. The Stem Cell Program also has an Influx high speed sterile cell sorter equipped
with four lasers and an automated cell deposition unit, an FC500 5-color flow cytometer, and a dedicated LSRII
cytometer for analysis. There are eight fully equipped tissue culture rooms associated with the shared translational
laboratory.
Animal: The principal investigator has three separate rooms within a contiguous suite, dedicated solely to the
breeding and care of immune deficient mice, in the Research III Barrier Facility on the UCD Medical Center
Campus, one block from the lab. There is a large laminar flow hood in each room. Two technicians care for the
mice, breeding and weaning, monitoring each cage twice daily, preparing antibiotic-treated water and irradiated
food, and changing microisolator cages twice a week. The health of the animals is checked daily. Sublethal
conditioning is done by irradiation, using the animal irradiator purchased for the Stem Cell Program, and located in
that facility, across the hall from the animal suite. Health screenings for each strain are done on a regular basis.
Computer: Hp Elitebook 2530p notebook PC laptop with docking station, CD writer, PDF writer, and 21” monitor
+ color printer, attached to University network and the Internet. Twelve similar PC computers and printers are
located in the main laboratory, and each student has a laptop. A graphics station permits generation of colored
posters, FACS and Microscopy file analysis, and blot scanning to generate photos for publications. Access to the
Internet, On-Line Journals, UC library – held publications, GenBank, etc. are freely available. Referencing is done
through the EndNote program linked into PubMed, and statistical analyses are done using the Excel software and a
program statistician is provided through the NIH-funded CTSC. The Dell laptop has a wireless card for internet
connection during travel, and the PI carries a blackberry for continual accessibility for the lab members.
Office: The P.I. has 200 square feet of non-shared office space in the new Institute for Regenerative Cures, that
opens directly into the lab. She is available to the students and technicians during the day. Her administrative
assistants and program managers have office space in the contiguous suites. GMP and clinical trial personnel are in a
suite next door to the translational lab. Our NIH-funded Clinical and Translational Science Center is next door to the
Institute, within the same building. Dr. Nolta’s private office has desk space and room for file cabinets, data
logbooks, data files, reference books, computer, phone with voicemail, printer, and room for meetings. The P.I. has
an additional computer, color printer/scanner/copier, and FAX in her home office. There are two conference rooms
for program use and a common eating area in the Institute for Regenerative Cures.