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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.


Related in: MedlinePlus

Levels of mRNA expression of proinflammatory cytokines are upregulated during adipogenic differentiation of ASCs. ASCs were grown in CCM and changed to ADM. Cells were harvested on days 7, 14, and 21 and analyzed by qRT-PCR. Data is normalized to undifferentiated cells. Mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001 relative to undifferentiated cells.
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fig4: Levels of mRNA expression of proinflammatory cytokines are upregulated during adipogenic differentiation of ASCs. ASCs were grown in CCM and changed to ADM. Cells were harvested on days 7, 14, and 21 and analyzed by qRT-PCR. Data is normalized to undifferentiated cells. Mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001 relative to undifferentiated cells.

Mentions: ASCs were expanded in CCM until confluent at which time ADM was added to the cells. Again, at 7, 14, and 21 days, cells were harvested and the mRNA expression of proinflammatory and anti-inflammatory cytokines was assessed relative to naive ASCs. ASCs induced with ADM for 7 days and 14 days demonstrated an increase in the expression of proinflammatory cytokines IL-12 (223.3-fold on day 7 and 4.3-fold on day 14, P < 0.05), interleukin-17 (IL-17; 69.8-fold on day 7 and 11.9-fold on day 14, P < 0.001), and ICAM-1 (8.4-fold on day 7 and 2.6-fold on day 14, P < 0.05, Figure 4(a)). In contrast, several proinflammatory cytokines demonstrated the highest levels of mRNA expression on day 21: mRNA levels for IL-6 were increased by 9.4-fold, IL-8 was increased by 929.5-fold, G-CSF was increased by 559.4-fold, CCL8 was increased by 163.8-fold, CXCL10 was increased by 43.9-fold, and TNF-α was increased by 139.5-fold (Figure 4(b), P < 0.001). In contrast, the mRNA expression of several proinflammatory cytokines was significantly reduced during adipogenic differentiation of ASCs: IL-1 (−4.2-fold on day 14 and −2.4-fold on day 21, P < 0.001), LIF (−5.8-fold on day 14, P < 0.001), and IFN-γ (−5.0-fold on day 14 and −3.6-fold on day 21, P < 0.001).


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)

Levels of mRNA expression of proinflammatory cytokines are upregulated during adipogenic differentiation of ASCs. ASCs were grown in CCM and changed to ADM. Cells were harvested on days 7, 14, and 21 and analyzed by qRT-PCR. Data is normalized to undifferentiated cells. Mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001 relative to undifferentiated cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4537750&req=5

fig4: Levels of mRNA expression of proinflammatory cytokines are upregulated during adipogenic differentiation of ASCs. ASCs were grown in CCM and changed to ADM. Cells were harvested on days 7, 14, and 21 and analyzed by qRT-PCR. Data is normalized to undifferentiated cells. Mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001 relative to undifferentiated cells.
Mentions: ASCs were expanded in CCM until confluent at which time ADM was added to the cells. Again, at 7, 14, and 21 days, cells were harvested and the mRNA expression of proinflammatory and anti-inflammatory cytokines was assessed relative to naive ASCs. ASCs induced with ADM for 7 days and 14 days demonstrated an increase in the expression of proinflammatory cytokines IL-12 (223.3-fold on day 7 and 4.3-fold on day 14, P < 0.05), interleukin-17 (IL-17; 69.8-fold on day 7 and 11.9-fold on day 14, P < 0.001), and ICAM-1 (8.4-fold on day 7 and 2.6-fold on day 14, P < 0.05, Figure 4(a)). In contrast, several proinflammatory cytokines demonstrated the highest levels of mRNA expression on day 21: mRNA levels for IL-6 were increased by 9.4-fold, IL-8 was increased by 929.5-fold, G-CSF was increased by 559.4-fold, CCL8 was increased by 163.8-fold, CXCL10 was increased by 43.9-fold, and TNF-α was increased by 139.5-fold (Figure 4(b), P < 0.001). In contrast, the mRNA expression of several proinflammatory cytokines was significantly reduced during adipogenic differentiation of ASCs: IL-1 (−4.2-fold on day 14 and −2.4-fold on day 21, P < 0.001), LIF (−5.8-fold on day 14, P < 0.001), and IFN-γ (−5.0-fold on day 14 and −3.6-fold on day 21, P < 0.001).

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.


Related in: MedlinePlus