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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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Related in: MedlinePlus

Osteogenesis-associated gene expression and protein secretion by human adipose-derived stromal cells (hASCs) and purified hASCs.A) There were no significant differences in the mRNA levels of Runt related transcription factor 2 (RUNX2), Osterix (OSX), Type I Collagen (COL1A1) or Osteocalcin (OCN) between hASCs and purified hASCs at 3, 7 or 14 days after osteogenic induction (P>0.05). B) There was no significant difference in OCN secretion between hASCs and purified hASCs at 24, 48 or 72 hours after osteogenic induction (P>0.05).
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pone-0056002-g004: Osteogenesis-associated gene expression and protein secretion by human adipose-derived stromal cells (hASCs) and purified hASCs.A) There were no significant differences in the mRNA levels of Runt related transcription factor 2 (RUNX2), Osterix (OSX), Type I Collagen (COL1A1) or Osteocalcin (OCN) between hASCs and purified hASCs at 3, 7 or 14 days after osteogenic induction (P>0.05). B) There was no significant difference in OCN secretion between hASCs and purified hASCs at 24, 48 or 72 hours after osteogenic induction (P>0.05).

Mentions: We examined the two cell types' expression of osteogenic genes such as Runt related transcription factor 2 (RUNX2), Osterix (OSX), Type I Collagen (COL1A1) and Osteocalcin (OCN) by real-time quantitative reverse transcription (qRT)-PCR. We found no significant differences (P>0.05) in the mRNA levels of these genes between hASCs and purified hASCs at 0, 3, 7 and 14 days after OI (Fig. 4A). The osteogenesis-associated secretion of OCN protein was also determined by radioimmunoassay. We found no significant difference (P>0.05) in OCN secretion between hASCs and purified hASCs at 24, 48 and 72 hours after OI (Fig. 4B).


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)

Osteogenesis-associated gene expression and protein secretion by human adipose-derived stromal cells (hASCs) and purified hASCs.A) There were no significant differences in the mRNA levels of Runt related transcription factor 2 (RUNX2), Osterix (OSX), Type I Collagen (COL1A1) or Osteocalcin (OCN) between hASCs and purified hASCs at 3, 7 or 14 days after osteogenic induction (P>0.05). B) There was no significant difference in OCN secretion between hASCs and purified hASCs at 24, 48 or 72 hours after osteogenic induction (P>0.05).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0056002-g004: Osteogenesis-associated gene expression and protein secretion by human adipose-derived stromal cells (hASCs) and purified hASCs.A) There were no significant differences in the mRNA levels of Runt related transcription factor 2 (RUNX2), Osterix (OSX), Type I Collagen (COL1A1) or Osteocalcin (OCN) between hASCs and purified hASCs at 3, 7 or 14 days after osteogenic induction (P>0.05). B) There was no significant difference in OCN secretion between hASCs and purified hASCs at 24, 48 or 72 hours after osteogenic induction (P>0.05).
Mentions: We examined the two cell types' expression of osteogenic genes such as Runt related transcription factor 2 (RUNX2), Osterix (OSX), Type I Collagen (COL1A1) and Osteocalcin (OCN) by real-time quantitative reverse transcription (qRT)-PCR. We found no significant differences (P>0.05) in the mRNA levels of these genes between hASCs and purified hASCs at 0, 3, 7 and 14 days after OI (Fig. 4A). The osteogenesis-associated secretion of OCN protein was also determined by radioimmunoassay. We found no significant difference (P>0.05) in OCN secretion between hASCs and purified hASCs at 24, 48 and 72 hours after OI (Fig. 4B).

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