Chondrogenesis of human bone marrow-derived mesenchymal stem cells in agarose culture.
Mesenchymal stem cells derived from human bone marrow (hBM-MSCs) can differentiate into chondrogenic cells for the potential treatment of injured articular cartilage. To evaluate agarose gels as a supportive material for chondrogenesis of hBM-MSCs, this study examined chondrogenesis of hBM-MSCs in the agarose cultures. Pellet cultures were employed to confirm the chondrogenic potential of the hBM-MSCs that were used in agarose cultures. The hBM-MSCs were seeded in 2% agarose constructs at the initial cell-seeding densities of 3, 6, and 9 x 10(6) cells/ml while each of pellets was formed using 2.5 x 10(5) cells. Chondrogenesis of hBM-MSCs was induced by culturing cell-agarose constructs and pellets for 21 days in the presence of a defined medium containing transforming growth factor beta3 (TGF-beta3). The analysis of reverse transcription-polymerase chain reaction showed that hBM-MSCs of agarose and pellet cultures expressed the chondrogenic markers of collagen type II and aggrecan in the presence of TGF-beta3. The deposition of cartilage-specific macromolecules was detected in both agarose and pellet cultures by histological and immunohistochemical assessments. Chondrogenesis of hBM-MSCs in agarose gels directly correlated with the initial cell-seeding density, with the cell-agarose constructs of higher initial cell-seeding density exhibiting more cartilage-specific gene expressions. This study establishes a basic model for future studies on chondrogenesis of hBM-MSCs using the agarose cultures.
Huang
C-Y Charles
CY
Research Service and Geriatrics Research, Education, and Clinical Center, Miami Veterans Affairs Medical Center, 1201 NW 16th Street, Miami, FL 33125, USA.
Reuben
Paul M
PM
D'Ippolito
Gianluca
G
Schiller
Paul C
PC
Cheung
Herman S
HS
D053674:Aggrecans; D001854:Bone Marrow Cells / Q000502:physiology*; D020219:Chondrogenesis / Q000502:physiology*; D024043:Collagen Type II / Q000378:metabolism; D003470:Culture Media; D017931:DNA Primers; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005240:Feasibility Studies; D006652:Histological Techniques; D006801:Humans; D007150:Immunohistochemistry; D037181:Lectins, C-Type; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D011509:Proteoglycans / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012685:Sepharose; D016212:Transforming Growth Factor beta; D053782:Transforming Growth Factor beta3
D053674:Aggrecans; D024043:Collagen Type II; D003470:Culture Media; D017931:DNA Primers; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D016212:Transforming Growth Factor beta; D053782:Transforming Growth Factor beta3; D012685:Sepharose