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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.

Show MeSH
Purification of human adipose-derived stromal cells (hASCs) by flow cytometric cell sorting.Purified hASCs positive for all four of the mesenchymal stem cell-specific markers CD44, CD73, CD90 and CD105 were isolated from hASCs (third passage) by flow cytometry.
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pone-0056002-g002: Purification of human adipose-derived stromal cells (hASCs) by flow cytometric cell sorting.Purified hASCs positive for all four of the mesenchymal stem cell-specific markers CD44, CD73, CD90 and CD105 were isolated from hASCs (third passage) by flow cytometry.

Mentions: Purified hASCs positive for all four of the surface markers CD44, CD73, CD90 and CD105 were isolated from P3 hASCs by FCCS (Fig. 2). The purified hASCs were then passaged twice before the following experiments were performed. Unpurified P3 hASCs were also passaged and cultured using the same methods as for the purified hASCs.


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)

Purification of human adipose-derived stromal cells (hASCs) by flow cytometric cell sorting.Purified hASCs positive for all four of the mesenchymal stem cell-specific markers CD44, CD73, CD90 and CD105 were isolated from hASCs (third passage) by flow cytometry.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0056002-g002: Purification of human adipose-derived stromal cells (hASCs) by flow cytometric cell sorting.Purified hASCs positive for all four of the mesenchymal stem cell-specific markers CD44, CD73, CD90 and CD105 were isolated from hASCs (third passage) by flow cytometry.
Mentions: Purified hASCs positive for all four of the surface markers CD44, CD73, CD90 and CD105 were isolated from P3 hASCs by FCCS (Fig. 2). The purified hASCs were then passaged twice before the following experiments were performed. Unpurified P3 hASCs were also passaged and cultured using the same methods as for the purified hASCs.

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