Procurement of Tissue Samples for Cell Cultures and Analyses

Background: It is now apparent that virtually every human post-natal tissue contains some
type of stem/progenitor cell population that is responsible for tissue turnover and repair.
Our previous studies have identified a subset of human bone marrow stromal cells (BMSCs) that
is multi-potent and able to regenerate bone, cartilage, myelosupportive stroma and
adipocytes, whereas stromal cells derived from human spleen (SpSCs) and thymus (TSCs) only
form myelosupportive stroma. Recent studies suggest that human adipose-derived stromal cells
(ASCs) may be similar to BMSCs, whereas stromal cells derived from human dermis (DSCs) have
none of these properties (and serve as a negative control for most experiments in vitro and
in vivo). How similar or dissimilar these different stromal cell populations are has yet to
be determined. Molecular profiling is needed to compare these different populations and in
order to elucidate the factors that control differentiation stromal cells into one cell type
or another.

Objectives: Surgical waste (bone with red marrow, dermis, spleen, thymus and adipose tissues)
from males and females of varying ages undergoing clinically indicated surgical procedures
will be used to establish stromal cell cultures to study the molecular profile and
differentiation capacity of the stromal cell populations from different tissues and to
further characterize the regulation of gene expression and protein synthesis in these
stem/progenitor cells.

Eligibility: Any patient who is undergoing clinically indicated surgery that entails removal
of bone with red marrow, dermis, spleen, thymus and adipose tissue, and who do not have: 1) a
history of metabolic bone disease, 2) any form of cancer or have been treated by chemotherapy
or irradiation for cancer, and 3) have not received agents known to affect bone metabolism.

Design: Normal surgical waste (bone with red marrow, dermis, spleen, thymus and adipose) from
procedures that are performed on males and females at Suburban Hospital will be placed in
nutrient medium (provided by NIDCR) and sent to the NIDCR for the establishment of cell
culture strains. Only the age, gender and site from which the tissue was removed will be
recorded. The cell cultures from the different tissues will be used to determine their
phenotypic character and differentiation properties by molecular profiling, and for studies
to elucidate the regulation of gene expression and protein synthesis. Similarly, a portion of
the samples obtained at Suburban Hospital will be used to generate histological sections of
the normal tissues that will be used to screen newly developed antibodies and molecular
probes. In some cases, RNA will be extracted from the tissues for RT-PCR analysis. These
samples (cells, sections, mRNA) will also serve as normal controls for studies performed on
samples obtained from patients with various diseases recruited to NIH under current NIH
protocols (97-D-0055, 97-D-0145, 01-D-0184).

- INCLUSION CRITERIA:

Any patient over the age of 18 years who is undergoing a clinically indicated or elective
surgery that entails removal of bone with red marrow,periosteum, dermis, spleen, thymus,
muscle and adipose tissue.

All patients must sign a document of informed consent indicating their understanding that
the specimens collected and demographic information provided is only for research purposes.

EXCLUSION CRITERIA:

Any patient that has a history of metabolic bone disease.

Any patient that has cancer of any type, or has been treated for cancer of any type.

Any patient that has been treated with a reagent known to affect bone metabolism
(bisphosphonates, steroids, vitamin D)