Cell Culture Facility by pgb16892


									                                      Cell Culture Facility
Kerry S. Campbell, Ph.D., Member, Director
Sharon D. Howard, B.A., HT-ASCP, Senior Scientific Associate
Joann R. Lutje, B.S., Scientific Technician
Dawn M. Riddle,* B.S., Scientific Technician
Sharon Connelly, Scientific Technician
A. Catherine Bluett, Technician
Laura Seeholzer, Student Assistant, Abington High School

Since it was established in 1981, the Cell Culture Facility has made it easier, more economical and more
effective to use mammalian cell cultures in research at Fox Chase. Cell culture procedures provide the
cellular materials for many of the molecular methods used to study normal and cancer cell proliferation,
cellular regulatory mechanisms, and normal and abnormal development. As a result, cell culture has
become a central technique in current molecular and cell biological research.
   The Cell Culture Facility provides a wide range of services and technical assistance that are tailored to
the individual needs of the user. The facility provides basic support services (preparation of numerous
culture media and a source of culture supplies and sterile reagents), cell propagation services, expertise in
establishing primary cultures from human and animal tissue samples, cell transformation services,
screening services for detecting mycoplasma contamination in cultured cells, consultation to assist in
experimental design, and sterile workstations for use by investigators who do not have facilities of their
own. A “nucleofection” service has become very popular among researchers to improve transfection effi-
ciencies in many mammalian cell lines. A total of 20 FCCC laboratories used this service in 2006 to per-
form 1463 transfections. A low oxygen incubator is also available for common use by Fox Chase
investigators. The cell culture facility is used by more than three-quarters of the Fox Chase laboratories,
illustrating the high demand for such technology and services at the Center.
   The following services were provided during the past year: 4327 liters of growth media and 2080 liters
of sterile reagents were prepared, 3200 vials of viable cells were banked in liquid nitrogen, and 702
samples were thawed for use. Mycoplasma screening was conducted on 315 samples from various labs at
Fox Chase, of which 57 were infected. The facility propagated 370 established cell lines and developed
an additional 19 primary cell lines, including 3 mouse mammary tumors, 8 embryonic mouse fibroblast
lines, one mouse kidney epithelial and fibroblast line, 2 human lung-angiomyolipomas, one angiomyoli-
poma, and 3 lipomas. The facility sent 2100 cell samples to other institutions and received 40 from out-
side sources. Two kidney fibroblast lines were propagated for extracellular matrix studies. Seven kidney
LAM and AML lines were expanded for injection into scid mice. Thirty liters of cells were maintained in
roller bottles and spinner cultures by the facility for antibody production or DNA and RNA preparation.
One hundred fifty cell lines were grown for harvesting DNA or RNA.
   Fetal bovine serum samples from 9 distributors were performance tested for the ability to promote
satisfactory growth rates of 6 sensitive cell lines. Based on these tests, the facility negotiated a reduced
price for bulk purchasing 1000 × 500 ml bottles of general serum from the optimal lot. Investigators can
purchase these optimal batches of serum at our reduced bulk acquisition cost. During the past year, more
than 900 × 500 ml bottles of fetal bovine serum were dispensed in this way.
   The Cell Culture Facility also provides technical support, cells, reagents, and supplies for establishing
germline transmissible embryonic stem (ES) cells for developing gene knockout mice. Seven different ES
cell lines from three strains of mice, as well as feeder layer cells (mouse embryo fibroblasts), and all
necessary reagents are in stock for immediate use in the facility. The facility has generated 400 vials of
irradiated mouse embryo fibroblasts for this purpose. During this same period, one ES cell transfection
project generated 371 neomycin-resistant clones. To date, 15 “knocked out” or “knocked in” ES cell lines
have been established by the facility. These genetically modified ES cell lines are subsequently injected
into blastocysts by the Transgenic Mouse Facility to establish chimeric mice bearing the modified gene.
This year 20 positive ES cell lines, developed at FCCC and other institutions were propagated for injection.
The Cell Culture Facility will continue to contribute substantial support to those efforts in the future.
   The Cell Culture Facility maintains a computer catalog that accounts for about 62,000 vials of cell lines
banked in liquid nitrogen within the facility for Fox Chase investigators. The computer database is accessi-
                             Fox Chase Cancer Center 2006 Scientific Report                   1
ble to researchers for searching via internal intranet access. This allows investigators to directly access their
inventory of frozen stocks and improve the sharing of cell reagents among the researchers at the Center.

* Personnel left Fox Chase

                               Fox Chase Cancer Center 2006 Scientific Report                    2

To top