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Unique responses of stem cell-derived vascular endothelial and mesenchymal cells to high levels of glucose.

Keats E, Khan ZA - PLoS ONE (2012)

Bottom Line: Our results show that high levels of glucose do not alter the derivation of either EPCs or MPCs.Interestingly, MPCs showed a transient reduction in growth upon glucose challenge.The findings further show that hyperglycemia may have detrimental effects on the MPCs, causing reduced growth and altering the differentiation potential.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, University of Western Ontario, London, Ontario, Canada.

ABSTRACT
Diabetes leads to complications in selected organ systems, and vascular endothelial cell (EC) dysfunction and loss is the key initiating and perpetuating step in the development of these complications. Experimental and clinical studies have shown that hyperglycemia leads to EC dysfunction in diabetes. Vascular stem cells that give rise to endothelial progenitor cells (EPCs) and mesenchymal progenitor cells (MPCs) represent an attractive target for cell therapy for diabetic patients. Whether these vascular stem/progenitor cells succumb to the adverse effects of high glucose remains unknown. We sought to determine whether adult vascular stem/progenitor cells display cellular activation and dysfunction upon exposure to high levels of glucose as seen in diabetic complications. Mononuclear cell fraction was prepared from adult blood and bone marrow. EPCs and MPCs were derived, characterized, and exposed to either normal glucose (5 mmol/L) or high glucose levels (25 mmol/L). We then assayed for cell activity and molecular changes following both acute and chronic exposure to high glucose. Our results show that high levels of glucose do not alter the derivation of either EPCs or MPCs. The adult blood-derived EPCs were also resistant to the effects of glucose in terms of growth. Acute exposure to high glucose levels increased caspase-3 activity in EPCs (1.4x increase) and mature ECs (2.3x increase). Interestingly, MPCs showed a transient reduction in growth upon glucose challenge. Our results also show that glucose skews the differentiation of MPCs towards the adipocyte lineage while suppressing other mesenchymal lineages. In summary, our studies show that EPCs are resistant to the effects of high levels of glucose, even following chronic exposure. The findings further show that hyperglycemia may have detrimental effects on the MPCs, causing reduced growth and altering the differentiation potential.

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Gene expression profiles of bmMPCs.Cells were cultured in 5 mmol/L (control) or 25 mmol/L (high glucose; HG) glucose for up to 14 days. RNA was isolated at day 1, day 3 (data not shown), and day 14 (data not shown) in order to assess changes in gene expression through qRT-PCR. (A) Oxidative stress marker expression in bmMPCs and uaSMCs showed no significant changes upon exposure to high levels of glucose. (B) HG induced an increase in the production of matrix proteins in mature uaSMCs, but caused downregulation of these same markers in the bmMPCs [*p<0.05 compared to control media; n = 3].
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pone-0038752-g004: Gene expression profiles of bmMPCs.Cells were cultured in 5 mmol/L (control) or 25 mmol/L (high glucose; HG) glucose for up to 14 days. RNA was isolated at day 1, day 3 (data not shown), and day 14 (data not shown) in order to assess changes in gene expression through qRT-PCR. (A) Oxidative stress marker expression in bmMPCs and uaSMCs showed no significant changes upon exposure to high levels of glucose. (B) HG induced an increase in the production of matrix proteins in mature uaSMCs, but caused downregulation of these same markers in the bmMPCs [*p<0.05 compared to control media; n = 3].

Mentions: We have previously shown altered cellular activity of ECs in high glucose conditions and in target organs of diabetic complications [24]–[26]. These altered activities include changes in the redox enzymes, expansion of extracellular matrix, and vasoactive factor alteration. We wanted to determine whether long-term culture of vascular progenitor cells would lead to similar alterations. Therefore, we cultured the cells in complete media (EBM-2/20% FBS with or without 25 mmol/L glucose) and performed a gene expression analyses. We profiled key cell activation genes (endothelin-1, ET-1; E-selectin; and intercellular adhesion molecule-1, ICAM-1), matrix protein genes (collagens 1,3,4; and fibronectin), and oxidative stress genes (catalase, Cat; glutathione peroxidase, GPx; heme oxygenase-1, HO-1; NADPH oxidase, p22 Phox; inducible nitric oxide synthase, NOS2; and superoxide dismutase-1, SOD-1). Interestingly, no significant effect of high levels of glucose was noted in endothelial cell activation, oxidative stress parameters, and matrix proteins at day 1, day 3, or day 14 (Figure 3). Analysis of bmMPCs also showed no changes in oxidative stress markers (Figure 4A). Significantly increased levels of matrix proteins (collagen 3, collagen 4, and fibronectin) in high glucose-treated uaSMCs were seen (Figure 4B). These same matrix protein mRNA levels were significantly reduced in the bmMPCs.


Unique responses of stem cell-derived vascular endothelial and mesenchymal cells to high levels of glucose.

Keats E, Khan ZA - PLoS ONE (2012)

Gene expression profiles of bmMPCs.Cells were cultured in 5 mmol/L (control) or 25 mmol/L (high glucose; HG) glucose for up to 14 days. RNA was isolated at day 1, day 3 (data not shown), and day 14 (data not shown) in order to assess changes in gene expression through qRT-PCR. (A) Oxidative stress marker expression in bmMPCs and uaSMCs showed no significant changes upon exposure to high levels of glucose. (B) HG induced an increase in the production of matrix proteins in mature uaSMCs, but caused downregulation of these same markers in the bmMPCs [*p<0.05 compared to control media; n = 3].
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3368917&req=5

pone-0038752-g004: Gene expression profiles of bmMPCs.Cells were cultured in 5 mmol/L (control) or 25 mmol/L (high glucose; HG) glucose for up to 14 days. RNA was isolated at day 1, day 3 (data not shown), and day 14 (data not shown) in order to assess changes in gene expression through qRT-PCR. (A) Oxidative stress marker expression in bmMPCs and uaSMCs showed no significant changes upon exposure to high levels of glucose. (B) HG induced an increase in the production of matrix proteins in mature uaSMCs, but caused downregulation of these same markers in the bmMPCs [*p<0.05 compared to control media; n = 3].
Mentions: We have previously shown altered cellular activity of ECs in high glucose conditions and in target organs of diabetic complications [24]–[26]. These altered activities include changes in the redox enzymes, expansion of extracellular matrix, and vasoactive factor alteration. We wanted to determine whether long-term culture of vascular progenitor cells would lead to similar alterations. Therefore, we cultured the cells in complete media (EBM-2/20% FBS with or without 25 mmol/L glucose) and performed a gene expression analyses. We profiled key cell activation genes (endothelin-1, ET-1; E-selectin; and intercellular adhesion molecule-1, ICAM-1), matrix protein genes (collagens 1,3,4; and fibronectin), and oxidative stress genes (catalase, Cat; glutathione peroxidase, GPx; heme oxygenase-1, HO-1; NADPH oxidase, p22 Phox; inducible nitric oxide synthase, NOS2; and superoxide dismutase-1, SOD-1). Interestingly, no significant effect of high levels of glucose was noted in endothelial cell activation, oxidative stress parameters, and matrix proteins at day 1, day 3, or day 14 (Figure 3). Analysis of bmMPCs also showed no changes in oxidative stress markers (Figure 4A). Significantly increased levels of matrix proteins (collagen 3, collagen 4, and fibronectin) in high glucose-treated uaSMCs were seen (Figure 4B). These same matrix protein mRNA levels were significantly reduced in the bmMPCs.

Bottom Line: Our results show that high levels of glucose do not alter the derivation of either EPCs or MPCs.Interestingly, MPCs showed a transient reduction in growth upon glucose challenge.The findings further show that hyperglycemia may have detrimental effects on the MPCs, causing reduced growth and altering the differentiation potential.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, University of Western Ontario, London, Ontario, Canada.

ABSTRACT
Diabetes leads to complications in selected organ systems, and vascular endothelial cell (EC) dysfunction and loss is the key initiating and perpetuating step in the development of these complications. Experimental and clinical studies have shown that hyperglycemia leads to EC dysfunction in diabetes. Vascular stem cells that give rise to endothelial progenitor cells (EPCs) and mesenchymal progenitor cells (MPCs) represent an attractive target for cell therapy for diabetic patients. Whether these vascular stem/progenitor cells succumb to the adverse effects of high glucose remains unknown. We sought to determine whether adult vascular stem/progenitor cells display cellular activation and dysfunction upon exposure to high levels of glucose as seen in diabetic complications. Mononuclear cell fraction was prepared from adult blood and bone marrow. EPCs and MPCs were derived, characterized, and exposed to either normal glucose (5 mmol/L) or high glucose levels (25 mmol/L). We then assayed for cell activity and molecular changes following both acute and chronic exposure to high glucose. Our results show that high levels of glucose do not alter the derivation of either EPCs or MPCs. The adult blood-derived EPCs were also resistant to the effects of glucose in terms of growth. Acute exposure to high glucose levels increased caspase-3 activity in EPCs (1.4x increase) and mature ECs (2.3x increase). Interestingly, MPCs showed a transient reduction in growth upon glucose challenge. Our results also show that glucose skews the differentiation of MPCs towards the adipocyte lineage while suppressing other mesenchymal lineages. In summary, our studies show that EPCs are resistant to the effects of high levels of glucose, even following chronic exposure. The findings further show that hyperglycemia may have detrimental effects on the MPCs, causing reduced growth and altering the differentiation potential.

Show MeSH
Related in: MedlinePlus