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Analysis of the Pro- and Anti-Inflammatory Cytokines Secreted by Adult Stem Cells during Differentiation.

Strong AL, Gimble JM, Bunnell BA - Stem Cells Int (2015)

Bottom Line: ASCs cultured in osteogenic and adipogenic differentiation medium were characterized by oil red o staining and alizarin red staining, respectively.ASCs undergoing osteogenic and adipogenic differentiation were isolated on days 7, 14, and 21 and assessed by qRT-PCR for the expression of pro- and anti-inflammatory cytokines.ASCs undergoing osteogenic differentiation expressed a distinct panel of cytokines that differed from the cytokine profile of ASCs undergoing adipogenic differentiation at each of the time points analyzed.

View Article: PubMed Central - PubMed

Affiliation: Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, 1430 Tulane Avenue, New Orleans, LA 70112, USA.

ABSTRACT
Adipose-derived stromal/stem cells (ASCs) are adult stem cells that have the potential to differentiate into mesenchymal lineage cells. The abundance of ASCs in adipose tissue and easy accessibility with relatively little donor site morbidity make them attractive candidate cells for tissue engineering and regenerative medicine. However, the underlying inflammatory process that occurs during ASC differentiation into adipocytes and osteoblast has not been extensively investigated. ASCs cultured in osteogenic and adipogenic differentiation medium were characterized by oil red o staining and alizarin red staining, respectively. ASCs undergoing osteogenic and adipogenic differentiation were isolated on days 7, 14, and 21 and assessed by qRT-PCR for the expression of pro- and anti-inflammatory cytokines. ASCs undergoing osteogenic differentiation expressed a distinct panel of cytokines that differed from the cytokine profile of ASCs undergoing adipogenic differentiation at each of the time points analyzed. Mapping the cytokine expression profile during ASC differentiation will provide insight into the role of inflammation in this process and identify potential targets that may aid in enhancing osteogenic or adipogenic differentiation for the purposes of tissue engineering and regenerative medicine.

No MeSH data available.


Schematic of ASCs undergoing osteogenic or adipogenic differentiation. Expression of proinflammatory and anti-inflammatory cytokines is dependent on the stage of differentiation.
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Related In: Results  -  Collection


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fig6: Schematic of ASCs undergoing osteogenic or adipogenic differentiation. Expression of proinflammatory and anti-inflammatory cytokines is dependent on the stage of differentiation.

Mentions: A quantitative comparison of the proinflammatory cytokines and anti-inflammatory cytokines expressed during osteogenic differentiation of ASCs demonstrates three distinct groups of cytokines. These groups of cytokines are categorized based on their induction at early, mid, and late stages of osteogenic differentiation (Figure 6). While the current study investigated the inflammatory cytokines secreted during the osteogenic differentiation of these cells in a cocktail of growth factors, others have taken the approach of treating progenitor cells with a similar cocktail of growth factors and supplemented the medium with additional inflammatory cytokines, such as TNF-α and IL-1 [11–14]. Most of these studies were conducted in bone marrow-derived mesenchymal stem cells (BMSCs), which are derived from the mesodermal lineage and have a similar differentiation potential as ASCs. Human BMSCs treated with TNF-α resulted in the activation of NF-κB, leading to increased mineralization and enhanced expression of osteogenic proteins, such as BMP2 and alkaline phosphatase, and transcription factors such as RUNX2 and Osterix [11, 12]. Human BMSCs treated with IL-1, likewise, enhanced differentiation into osteoblasts through the Wnt-5a/receptor tyrosine kinase-like orphan receptor 2 pathway [13]. Ferreira et al. found that IL-1 also enhanced mineralization through both nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and mitogen-activated protein kinase (MAPK) pathways [14]. The results presented in the current study are consistent with previously published reports showing an upregulation of TNF-α and IL-1 during differentiation. The present study supplements the current body of literature and highlights specific inflammatory factors that should be investigated further based on their induction during osteogenic differentiation. Additional studies, however, are necessary to elucidate the precise mechanism by which these cytokines effect osteogenic differentiation.


Analysis of the Pro- and Anti-Inflammatory Cytokines Secreted by Adult Stem Cells during Differentiation.

Strong AL, Gimble JM, Bunnell BA - Stem Cells Int (2015)

Schematic of ASCs undergoing osteogenic or adipogenic differentiation. Expression of proinflammatory and anti-inflammatory cytokines is dependent on the stage of differentiation.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4537750&req=5

fig6: Schematic of ASCs undergoing osteogenic or adipogenic differentiation. Expression of proinflammatory and anti-inflammatory cytokines is dependent on the stage of differentiation.
Mentions: A quantitative comparison of the proinflammatory cytokines and anti-inflammatory cytokines expressed during osteogenic differentiation of ASCs demonstrates three distinct groups of cytokines. These groups of cytokines are categorized based on their induction at early, mid, and late stages of osteogenic differentiation (Figure 6). While the current study investigated the inflammatory cytokines secreted during the osteogenic differentiation of these cells in a cocktail of growth factors, others have taken the approach of treating progenitor cells with a similar cocktail of growth factors and supplemented the medium with additional inflammatory cytokines, such as TNF-α and IL-1 [11–14]. Most of these studies were conducted in bone marrow-derived mesenchymal stem cells (BMSCs), which are derived from the mesodermal lineage and have a similar differentiation potential as ASCs. Human BMSCs treated with TNF-α resulted in the activation of NF-κB, leading to increased mineralization and enhanced expression of osteogenic proteins, such as BMP2 and alkaline phosphatase, and transcription factors such as RUNX2 and Osterix [11, 12]. Human BMSCs treated with IL-1, likewise, enhanced differentiation into osteoblasts through the Wnt-5a/receptor tyrosine kinase-like orphan receptor 2 pathway [13]. Ferreira et al. found that IL-1 also enhanced mineralization through both nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and mitogen-activated protein kinase (MAPK) pathways [14]. The results presented in the current study are consistent with previously published reports showing an upregulation of TNF-α and IL-1 during differentiation. The present study supplements the current body of literature and highlights specific inflammatory factors that should be investigated further based on their induction during osteogenic differentiation. Additional studies, however, are necessary to elucidate the precise mechanism by which these cytokines effect osteogenic differentiation.

Bottom Line: ASCs cultured in osteogenic and adipogenic differentiation medium were characterized by oil red o staining and alizarin red staining, respectively.ASCs undergoing osteogenic and adipogenic differentiation were isolated on days 7, 14, and 21 and assessed by qRT-PCR for the expression of pro- and anti-inflammatory cytokines.ASCs undergoing osteogenic differentiation expressed a distinct panel of cytokines that differed from the cytokine profile of ASCs undergoing adipogenic differentiation at each of the time points analyzed.

View Article: PubMed Central - PubMed

Affiliation: Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, 1430 Tulane Avenue, New Orleans, LA 70112, USA.

ABSTRACT
Adipose-derived stromal/stem cells (ASCs) are adult stem cells that have the potential to differentiate into mesenchymal lineage cells. The abundance of ASCs in adipose tissue and easy accessibility with relatively little donor site morbidity make them attractive candidate cells for tissue engineering and regenerative medicine. However, the underlying inflammatory process that occurs during ASC differentiation into adipocytes and osteoblast has not been extensively investigated. ASCs cultured in osteogenic and adipogenic differentiation medium were characterized by oil red o staining and alizarin red staining, respectively. ASCs undergoing osteogenic and adipogenic differentiation were isolated on days 7, 14, and 21 and assessed by qRT-PCR for the expression of pro- and anti-inflammatory cytokines. ASCs undergoing osteogenic differentiation expressed a distinct panel of cytokines that differed from the cytokine profile of ASCs undergoing adipogenic differentiation at each of the time points analyzed. Mapping the cytokine expression profile during ASC differentiation will provide insight into the role of inflammation in this process and identify potential targets that may aid in enhancing osteogenic or adipogenic differentiation for the purposes of tissue engineering and regenerative medicine.

No MeSH data available.