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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

Osteogenic differentiation of ASCs increases expression of anti-inflammatory cytokines. ASCs were grown in CCM and then changed to ODM. After 7, 14, and 21 days, cells were harvested 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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fig3: Osteogenic differentiation of ASCs increases expression of anti-inflammatory cytokines. ASCs were grown in CCM and then changed to ODM. After 7, 14, and 21 days, cells were harvested 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: The expression of anti-inflammatory cytokines was also assessed during osteogenic differentiation. Of the 14 anti-inflammatory cytokines assessed, seven genes were significantly upregulated in ASCs following culture in ODM for 7 days, which diminished by days 14 and 21 after induction. Increased expression of interleukin-10 (IL-10; 122.8-fold, P < 0.001), interleukin-13 (IL-13; 59.8-fold, P < 0.001), interleukin-18 binding protein (IL-18BP; 38.9-fold, P < 0.001), interleukin-35 (IL-35; 102.7-fold, P < 0.001), chemokine (C-C motif) ligand 2 (CCL2; 161.2-fold, P < 0.001), cyclooxygenase 2 (COX2; 614.4-fold, P < 0.001), and stanniocalcin 1 (STC-1; 230.8-fold, P < 0.001, Figure 3(a)) was observed. In contrast, ASCs cultured in ODM for 7 and 14 days demonstrated a sustained increase in the expression of interleukin-1 receptor antagonist (IL-1RA; 2617.7-fold on day 7 and 3024.7-fold on day 14, P < 0.001) and tumor necrosis factor-stimulated gene 6 (TSG-6; 51.2-fold on day 7 and 65.2-fold on day 14, P < 0.001; Figure 3(b)). Cells cultured in ODM demonstrated a biphasic induction in mRNA expression of anti-inflammatory cytokine interleukin-11 (IL-11; 4.5-fold on day 7 and 22.9-fold on day 21, P < 0.001), tumor necrosis factor receptor superfamily member (TNFRSF1A; 4.5-fold on day 7 and 3.5-fold on day 21, P < 0.001), prostaglandin E synthase 2 (PTGES2; 5.7-fold on day 7 and 5.5-fold on day 21, P < 0.001), and TGF-β (17.0-fold on day 7 and 28.0-fold on day 21, P < 0.001; Figure 3(c)). In contrast, mRNA expression of hepatocyte growth factor (HGF) was reduced by −2.9-fold, −50.0-fold, and −7.1-fold on days 7, 14, and 21, respectively (P < 0.001; Figure 3(d)).


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)

Osteogenic differentiation of ASCs increases expression of anti-inflammatory cytokines. ASCs were grown in CCM and then changed to ODM. After 7, 14, and 21 days, cells were harvested 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

fig3: Osteogenic differentiation of ASCs increases expression of anti-inflammatory cytokines. ASCs were grown in CCM and then changed to ODM. After 7, 14, and 21 days, cells were harvested 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: The expression of anti-inflammatory cytokines was also assessed during osteogenic differentiation. Of the 14 anti-inflammatory cytokines assessed, seven genes were significantly upregulated in ASCs following culture in ODM for 7 days, which diminished by days 14 and 21 after induction. Increased expression of interleukin-10 (IL-10; 122.8-fold, P < 0.001), interleukin-13 (IL-13; 59.8-fold, P < 0.001), interleukin-18 binding protein (IL-18BP; 38.9-fold, P < 0.001), interleukin-35 (IL-35; 102.7-fold, P < 0.001), chemokine (C-C motif) ligand 2 (CCL2; 161.2-fold, P < 0.001), cyclooxygenase 2 (COX2; 614.4-fold, P < 0.001), and stanniocalcin 1 (STC-1; 230.8-fold, P < 0.001, Figure 3(a)) was observed. In contrast, ASCs cultured in ODM for 7 and 14 days demonstrated a sustained increase in the expression of interleukin-1 receptor antagonist (IL-1RA; 2617.7-fold on day 7 and 3024.7-fold on day 14, P < 0.001) and tumor necrosis factor-stimulated gene 6 (TSG-6; 51.2-fold on day 7 and 65.2-fold on day 14, P < 0.001; Figure 3(b)). Cells cultured in ODM demonstrated a biphasic induction in mRNA expression of anti-inflammatory cytokine interleukin-11 (IL-11; 4.5-fold on day 7 and 22.9-fold on day 21, P < 0.001), tumor necrosis factor receptor superfamily member (TNFRSF1A; 4.5-fold on day 7 and 3.5-fold on day 21, P < 0.001), prostaglandin E synthase 2 (PTGES2; 5.7-fold on day 7 and 5.5-fold on day 21, P < 0.001), and TGF-β (17.0-fold on day 7 and 28.0-fold on day 21, P < 0.001; Figure 3(c)). In contrast, mRNA expression of hepatocyte growth factor (HGF) was reduced by −2.9-fold, −50.0-fold, and −7.1-fold on days 7, 14, and 21, respectively (P < 0.001; Figure 3(d)).

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