49.
Collagen-Coated Polytetrafluoroethane Membrane Inserts Enhances
Chondrogenic Differentiation of Human Cord Blood Multi-Lineage
Progenitor Cells
Samir Munir, Kjeld Søballe, Michael Ulrich-Vinther, Thomas Gadegaard
Koch, Rita A. Kandel, W. David Lee
Orthopedic Research Laboratory Aarhus University Hospital; Orthopedic
Research Laboratory, Aarhus University Hospital; Orthopedic Research
Laboratory, Aarhus University Hospital; Department of Biomedical Sciences,
Ontario Veterinary College, Guelph, Canada; Department of Pathology and
Laboratory Medicine, Mount Sinai Hospital, Toronto, Canada; Department of
Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, Canada
Background:
Articular chondrocytes and bone marrow-derived multipotent
mesenchymal stromal cells (MSCs) are the favoured cells for cartilage tissue
engineering. Umbilical cord blood has proven an alternative source of MSCs
and moreover they may be more potent chondroprogenitor cells than bone-
marrow MSCs.
Purpose / Aim of Study:
Multilineage progenitor cells (MLPCs) are clonal
cord blood-derived MSCs and may therefore provide a cell source with more
reproducible outcomes compared to heterogeneous primary MSC cultures.
Materials and Methods:
We evaluated the chondrogenic potency of MLPCs
in standard micromass pellet system, layered on calcium polyphosphate (CPP),
and on semi-permeable polytetrafluoroethane membranes with and without
collagen type I, II or IV pre-coating.
Findings / Results:
The MPLC cell line used in this study possessed poor
chondrogenic potency overall, but membrane culturing resulted in a
multicellular layer tissue with formation of more cartilaginous tissue compared
to micromass or CPP culture. In the membrane system MLPCs produced
pellucid discs, 12 mm in diameter by 1 mm in thickness from 2x10^6 cells.
The discs had hyaline-like cartilage extracellular matrix, with 4-fold greater
proteoglycan content compared to MLPCs differentiated in standard
micromass pellet cultures. The expression of cartilage specific genes for
aggrecan, collagen II and SOX9 was significantly increased in uncoated as
well as collagen type I and IV coated membrane cultures compared with
micromass or CPP cultures.
Conclusions:
In conclusion, we demonstrate that MLPCs possess’
chondrogenic potency, which increased when cultured scaffold-free on
membrane inserts resulting in multicellular-layered hyaline-like cartilage
tissue. Evaluating the effect of culturing pre-differentiated MLPCs on CPP is
an obvious next step since direct seeding of MLPCs on CPP did not yield
satisfactory biphasic constructs.
