Bone formation in vivo: comparison of osteogenesis by transplanted mouse and human marrow stromal fibroblasts.
BACKGROUND
Marrow stromal fibroblasts (MSFs) are known to contain bone precursor cells. However, the osteogenic potential of human MSFs has been poorly characterized. The aim of this study was to compare the osteogenic capacity of mouse and human MSFs after implantation in vivo.


METHODS
After in vitro expansion, MSFs were loaded into a number of different vehicles and transplanted subcutaneously into immunodeficient mice.


RESULTS
Mouse MSFs transplanted within gelatin, polyvinyl sponges, and collagen matrices all formed a capsule of cortical-like bone surrounding a cavity with active hematopoiesis. In transplants of MSFs from transgenic mice harboring type I procollagen-chloramphenicol acetyltransferase constructs, chloramphenicol acetyltransferase activity was maintained for up to 14 weeks, indicating prolonged bone formation by transplanted MSFs. New bone formation by human MSFs was more dependent on both the in vitro expansion conditions and transplantation vehicles. Within gelatin, woven bone was observed sporadically and only after culture in the presence of dexamethasone and L-ascorbic acid phosphate magnesium salt n-hydrate. Consistent bone formation by human MSFs was achieved only within vehicles containing hydroxyapatite/tricalcium phosphate ceramics (HA/TCP) in the form of blocks, powder, and HA/TCP powder-type I bovine fibrillar collagen strips, and bone was maintained for at least 19 weeks. Cells of the new bone were positive for human osteonectin showing their donor origin. HA/TCP powder, the HA/TCP powder-type I bovine fibrillar collagen strips, and HA/TCP powder held together with fibrin were easier to load and supported more extensive osteogenesis than HA/TCP blocks and thus may be more applicable for therapeutic use.


CONCLUSIONS
In this article, we describe the differences in the requirements for mouse and human MSFs to form bone, and report the development of a methodology for the consistent in vivo generation of extensive bone from human MSFs.
Krebsbach
P H
PH
Laboratory of Developmental Biology and Bone Research Branch, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland 20892, USA.
Kuznetsov
S A
SA
Satomura
K
K
Emmons
R V
RV
Rowe
D W
DW
Robey
P G
PG
D000818:Animals; D001846:Bone Development / Q000502:physiology*; D001854:Bone Marrow Cells; D002130:Calcium Phosphates; D017690:Cell Transplantation / Q000379:methods; D002478:Cells, Cultured; D002516:Ceramics; D002648:Child; D002675:Child, Preschool; D003094:Collagen; D017886:Durapatite; D005260:Female; D005347:Fibroblasts / Q000637:transplantation; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D010012:Osteogenesis / Q000502:physiology; D011145:Polyvinyls; D011161:Porifera; D014019:Tissue Donors
D002130:Calcium Phosphates; D011145:Polyvinyls; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; C485838:calcium phosphate, monobasic, anhydrous; D003094:Collagen; D017886:Durapatite; C020243:calcium phosphate; C485829:calcium phosphate, dibasic, anhydrous