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Flow cytometric cell sorting and in vitro pre-osteoinduction are not requirements for in vivo bone formation by human adipose-derived stromal cells.

Liu Y, Zhao Y, Zhang X, Chen T, Zhao X, Ma GE, Zhou Y - PLoS ONE (2013)

Bottom Line: We found that pre-OI enhanced the in vitro osteogenic capacity of hASCs.Interestingly, we also found that purified hASCs had an osteogenic potential similar to that of unpurified hASCs in vitro and in vivo.These results suggest that FCCS and in vitro pre-OI are not requirements for in vivo bone formation by hASCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Prosthodontics, School and Hospital of Stomatology, Peking University, Beijing, China.

ABSTRACT
Human adipose-derived stromal cells (hASCs) are a promising cell source for bone tissue engineering. However, before the clinical application of hASCs for the treatment of bone defects, key questions require answers, including whether pre-osteoinduction (OI) and flow cytometric cell purification are indispensible steps for in vivo bone formation by hASCs. In this study, hASCs were purified by flow cytometric cell sorting (FCCS). The osteogenic capabilities of hASCs and purified hASCs with or without pre-osteoinduction were examined through in vitro and in vivo experiments. We found that pre-OI enhanced the in vitro osteogenic capacity of hASCs. However, 8 weeks after in vivo implantation, there were no significant differences between hASCs and hASCs that had undergone OI (hASCs+OI) or between purified hASCs and purified hASCs+OI (P>0.05). Interestingly, we also found that purified hASCs had an osteogenic potential similar to that of unpurified hASCs in vitro and in vivo. These results suggest that FCCS and in vitro pre-OI are not requirements for in vivo bone formation by hASCs.

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In vivo bone formation by human adipose-derived stromal cells (hASCs).A) Gross observation and soft X-ray examination showed that hASCs, purified hASCs, hASCs that had undergone osteoinduction (OI) (hASCs+OI) and purified hASCs+OI formed bone-like tissues of relatively higher density than blank controls and fibroblast controls. B) Hematoxylin and eosin staining showed bone-like tissues with the typical structure of osteocyte lacunae in all four groups 4 weeks after implantation. The scale bar represents 50 µm. C) Eight weeks after implantation, the area of bone formation was larger in all four groups. The scale bar represents 50 µm. D) Quantitative measurements of bone-like tissues demonstrated that, 4 weeks after implantation, the area of bone formation was significantly increased in hASCs+OI and purified hASCs+OI compared with hASCs and purified hASCs without OI. However, 8 weeks after implantation there were no significant differences among the four groups. *P<0.05 compared with hASCs without OI; #P<0.05 compared with purified hASCs without OI.
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pone-0056002-g006: In vivo bone formation by human adipose-derived stromal cells (hASCs).A) Gross observation and soft X-ray examination showed that hASCs, purified hASCs, hASCs that had undergone osteoinduction (OI) (hASCs+OI) and purified hASCs+OI formed bone-like tissues of relatively higher density than blank controls and fibroblast controls. B) Hematoxylin and eosin staining showed bone-like tissues with the typical structure of osteocyte lacunae in all four groups 4 weeks after implantation. The scale bar represents 50 µm. C) Eight weeks after implantation, the area of bone formation was larger in all four groups. The scale bar represents 50 µm. D) Quantitative measurements of bone-like tissues demonstrated that, 4 weeks after implantation, the area of bone formation was significantly increased in hASCs+OI and purified hASCs+OI compared with hASCs and purified hASCs without OI. However, 8 weeks after implantation there were no significant differences among the four groups. *P<0.05 compared with hASCs without OI; #P<0.05 compared with purified hASCs without OI.

Mentions: To compare in vivo bone formation capabilities and to determine the function of in vitro pre-OI, hASCs, purified hASCs, hASCs+OI and purified hASCs+OI were transplanted subcutaneously into nude mice along with β-tricalcium phosphate (β-TCP). Blank controls and fibroblast controls were used in this experiment. Gross observation and soft X-ray examination showed that hASCs, purified hASCs, hASCs+OI and purified hASCs+OI could all form bone-like tissues with a relatively higher density than blank controls and fibroblast controls (Fig. 6A).


Flow cytometric cell sorting and in vitro pre-osteoinduction are not requirements for in vivo bone formation by human adipose-derived stromal cells.

Liu Y, Zhao Y, Zhang X, Chen T, Zhao X, Ma GE, Zhou Y - PLoS ONE (2013)

In vivo bone formation by human adipose-derived stromal cells (hASCs).A) Gross observation and soft X-ray examination showed that hASCs, purified hASCs, hASCs that had undergone osteoinduction (OI) (hASCs+OI) and purified hASCs+OI formed bone-like tissues of relatively higher density than blank controls and fibroblast controls. B) Hematoxylin and eosin staining showed bone-like tissues with the typical structure of osteocyte lacunae in all four groups 4 weeks after implantation. The scale bar represents 50 µm. C) Eight weeks after implantation, the area of bone formation was larger in all four groups. The scale bar represents 50 µm. D) Quantitative measurements of bone-like tissues demonstrated that, 4 weeks after implantation, the area of bone formation was significantly increased in hASCs+OI and purified hASCs+OI compared with hASCs and purified hASCs without OI. However, 8 weeks after implantation there were no significant differences among the four groups. *P<0.05 compared with hASCs without OI; #P<0.05 compared with purified hASCs without OI.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3569421&req=5

pone-0056002-g006: In vivo bone formation by human adipose-derived stromal cells (hASCs).A) Gross observation and soft X-ray examination showed that hASCs, purified hASCs, hASCs that had undergone osteoinduction (OI) (hASCs+OI) and purified hASCs+OI formed bone-like tissues of relatively higher density than blank controls and fibroblast controls. B) Hematoxylin and eosin staining showed bone-like tissues with the typical structure of osteocyte lacunae in all four groups 4 weeks after implantation. The scale bar represents 50 µm. C) Eight weeks after implantation, the area of bone formation was larger in all four groups. The scale bar represents 50 µm. D) Quantitative measurements of bone-like tissues demonstrated that, 4 weeks after implantation, the area of bone formation was significantly increased in hASCs+OI and purified hASCs+OI compared with hASCs and purified hASCs without OI. However, 8 weeks after implantation there were no significant differences among the four groups. *P<0.05 compared with hASCs without OI; #P<0.05 compared with purified hASCs without OI.
Mentions: To compare in vivo bone formation capabilities and to determine the function of in vitro pre-OI, hASCs, purified hASCs, hASCs+OI and purified hASCs+OI were transplanted subcutaneously into nude mice along with β-tricalcium phosphate (β-TCP). Blank controls and fibroblast controls were used in this experiment. Gross observation and soft X-ray examination showed that hASCs, purified hASCs, hASCs+OI and purified hASCs+OI could all form bone-like tissues with a relatively higher density than blank controls and fibroblast controls (Fig. 6A).

Bottom Line: We found that pre-OI enhanced the in vitro osteogenic capacity of hASCs.Interestingly, we also found that purified hASCs had an osteogenic potential similar to that of unpurified hASCs in vitro and in vivo.These results suggest that FCCS and in vitro pre-OI are not requirements for in vivo bone formation by hASCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Prosthodontics, School and Hospital of Stomatology, Peking University, Beijing, China.

ABSTRACT
Human adipose-derived stromal cells (hASCs) are a promising cell source for bone tissue engineering. However, before the clinical application of hASCs for the treatment of bone defects, key questions require answers, including whether pre-osteoinduction (OI) and flow cytometric cell purification are indispensible steps for in vivo bone formation by hASCs. In this study, hASCs were purified by flow cytometric cell sorting (FCCS). The osteogenic capabilities of hASCs and purified hASCs with or without pre-osteoinduction were examined through in vitro and in vivo experiments. We found that pre-OI enhanced the in vitro osteogenic capacity of hASCs. However, 8 weeks after in vivo implantation, there were no significant differences between hASCs and hASCs that had undergone OI (hASCs+OI) or between purified hASCs and purified hASCs+OI (P>0.05). Interestingly, we also found that purified hASCs had an osteogenic potential similar to that of unpurified hASCs in vitro and in vivo. These results suggest that FCCS and in vitro pre-OI are not requirements for in vivo bone formation by hASCs.

Show MeSH
Related in: MedlinePlus