Incipient analysis of mesenchymal stem-cell-derived osteogenesis.
Tissue regeneration strategies invoke cell-based therapies for effective tissue formation. Current assessment of mesenchymal stem cell (MSC) directed bone regeneration during in vivo assays is dependent on histologic determination of bone formation. It was the aim of this study to determine the relationship between bone sialoprotein (BSP) expression and osteocalcin expression with subsequent osteogenesis occurring in MSC-based implants. RT-PCR assessment of human actin, collagen type I, BSP, and osteocalcin indicated that undifferentiated cells did not express BSP or osteocalcin. Three weeks following implantation, human BSP could be identified in RNAs isolated from the retrieved implants. For every implant from which human BSP cDNA was amplified, parallel implants harvested at 6 weeks demonstrated bone formation at the histologic level. This study confirms that, in the context of the severe combined immunodeficiency disease (SCID) mouse model, culture-expanded, cryopreserved human MSCs have osteogenic potential and demonstrates that implanted cell gene expression can reveal the early onset of bone formation.
Cooper
L F
LF
1 Dental Research Center, University of North Carolina School of Dentistry, Chapel Hill, 27559-7450, USA. Lyndon_Cooper@Dentistry.unc.edu
Harris
C T
CT
Bruder
S P
SP
Kowalski
R
R
Kadiyala
S
S
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D016026:Bone Marrow Transplantation; D001861:Bone Regeneration / Q000235:genetics / Q000502:physiology*; D003114:Colony-Forming Units Assay; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D058574:Integrin-Binding Sialoprotein; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000166:cytology; D015675:Osteocalcin / Q000096:biosynthesis / Q000235:genetics; D010012:Osteogenesis / Q000235:genetics* / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012795:Sialoglycoproteins / Q000096:biosynthesis / Q000235:genetics; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*
C546871:IBSP protein, human; C546872:Ibsp protein, mouse; D058574:Integrin-Binding Sialoprotein; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D015675:Osteocalcin