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Hyperglycemic Stress Impairs the Stemness Capacity of Kidney Stem Cells in Rats.

Yang G, Jia Y, Li C, Cheng Q, Yue W, Pei X - PLoS ONE (2015)

Bottom Line: However, KSC proliferation, differentiation ability and tolerance to hypoxia were decreased in high-glucose cultures.Taken together, these results suggest the high-glucose microenvironment can damage the reparative ability of KSCs.It may result in a decreased of recovery capability of renal tubules from injury.

View Article: PubMed Central - PubMed

Affiliation: Department of Geriatric Nephrology, Chinese PLA General Hospital, State Key Laboratory of Kidney Disease, Beijing, China.

ABSTRACT
The incidence of acute kidney injury in patients with diabetes is significantly higher than that of patients without diabetes, and may be associated with the poor stemness capacity of kidney stem cells (KSCs) and limited recovery of injured renal tubules. To investigate the effects of hyperglycemic stress on KSC stemness, KSCs were isolated from the rat renal papilla and analyzed for their self-renewal and differentiation abilities. Our results showed that isolated KSCs expressed the mesenchymal stem cell markers N-cadherin, Nestin, CD133, CD29, CD90, and CD73. Moreover, KSCs co-cultured with hypoxia-injured renal tubular epithelial cell (RTECs) induced the expression of the mature epithelial cell marker CK18, suggesting that the KSCs could differentiate into RTECs in vitro. However, KSC proliferation, differentiation ability and tolerance to hypoxia were decreased in high-glucose cultures. Taken together, these results suggest the high-glucose microenvironment can damage the reparative ability of KSCs. It may result in a decreased of recovery capability of renal tubules from injury.

No MeSH data available.


Related in: MedlinePlus

Schematic of the KSC epithelial induction protocol.KSC epithelial differentiation was determined by Transwell co-culture differentiation assay. Hypoxia-injured RTECs and KSCs were prepared and plated in the Transwell inserts and wells, respectively. Cells were co-cultured in induction media for three days, moved to renal epithelial cell growth medium (REGM) for two days, and then changed back to induction medium. This series was repeated twice for a total 10 days. The inserts containing hypoxia-injured RTECs were renewed every three days.
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pone.0139607.g001: Schematic of the KSC epithelial induction protocol.KSC epithelial differentiation was determined by Transwell co-culture differentiation assay. Hypoxia-injured RTECs and KSCs were prepared and plated in the Transwell inserts and wells, respectively. Cells were co-cultured in induction media for three days, moved to renal epithelial cell growth medium (REGM) for two days, and then changed back to induction medium. This series was repeated twice for a total 10 days. The inserts containing hypoxia-injured RTECs were renewed every three days.

Mentions: Epithelial KSC differentiation was evaluated by Transwell co-culture differentiation assay (0.4 μm membrane diameter, Corning Inc., USA). First, hypoxia-injured RTECs (2 × 103 RTECs plated in Transwell inserts) were prepared using the following sequential culture protocol: (1) DMEM/F12/10% FBS media in a 21% O2/5% CO2 atmosphere for 24 h; (2) DMEM/F12/1% FBS in a 1% O2/5% CO2 atmosphere incubator (Heraeus, Germany) for 4 h; (3) DMEM/F12/10% FCS in a 5% CO2 atmosphere for 2 h; (4) renal epithelial cell growth medium (REGM) (Lonza, Switzerland) and 21% O2/5% CO2 atmosphere for 24 h. Second, the KSCs were plated at 104 cells per well into 24-well plates. Co-cultures were then performed by transferring the inserts containing injured RTECs into the KSCs wells and induction medium, including 10 ng/mL activin A (PeproTech), 10 ng/mL BMP-7 (PeproTech), and 5 μM of retinoic acid (Sigma, USA), was added. Three days later, the culture media was replaced with REGM for two days, and then changed into induction medium again. This sequence was repeated twice for a total 10 days. The inserts containing hypoxia-injured RTECs were renewed every three days. Following the immunofluorescence protocol described above, KSCs were stained with mouse anti-CK18 polyclonal antibody (Abcam, UK), mouse anti-E-cadherin polyclonal antibody, and rabbit anti-ZO-1 polyclonal antibody (Santa Cruz, USA). The induction strategy is shown in Fig 1.


Hyperglycemic Stress Impairs the Stemness Capacity of Kidney Stem Cells in Rats.

Yang G, Jia Y, Li C, Cheng Q, Yue W, Pei X - PLoS ONE (2015)

Schematic of the KSC epithelial induction protocol.KSC epithelial differentiation was determined by Transwell co-culture differentiation assay. Hypoxia-injured RTECs and KSCs were prepared and plated in the Transwell inserts and wells, respectively. Cells were co-cultured in induction media for three days, moved to renal epithelial cell growth medium (REGM) for two days, and then changed back to induction medium. This series was repeated twice for a total 10 days. The inserts containing hypoxia-injured RTECs were renewed every three days.
© Copyright Policy
Related In: Results  -  Collection

License
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getmorefigures.php?uid=PMC4592017&req=5

pone.0139607.g001: Schematic of the KSC epithelial induction protocol.KSC epithelial differentiation was determined by Transwell co-culture differentiation assay. Hypoxia-injured RTECs and KSCs were prepared and plated in the Transwell inserts and wells, respectively. Cells were co-cultured in induction media for three days, moved to renal epithelial cell growth medium (REGM) for two days, and then changed back to induction medium. This series was repeated twice for a total 10 days. The inserts containing hypoxia-injured RTECs were renewed every three days.
Mentions: Epithelial KSC differentiation was evaluated by Transwell co-culture differentiation assay (0.4 μm membrane diameter, Corning Inc., USA). First, hypoxia-injured RTECs (2 × 103 RTECs plated in Transwell inserts) were prepared using the following sequential culture protocol: (1) DMEM/F12/10% FBS media in a 21% O2/5% CO2 atmosphere for 24 h; (2) DMEM/F12/1% FBS in a 1% O2/5% CO2 atmosphere incubator (Heraeus, Germany) for 4 h; (3) DMEM/F12/10% FCS in a 5% CO2 atmosphere for 2 h; (4) renal epithelial cell growth medium (REGM) (Lonza, Switzerland) and 21% O2/5% CO2 atmosphere for 24 h. Second, the KSCs were plated at 104 cells per well into 24-well plates. Co-cultures were then performed by transferring the inserts containing injured RTECs into the KSCs wells and induction medium, including 10 ng/mL activin A (PeproTech), 10 ng/mL BMP-7 (PeproTech), and 5 μM of retinoic acid (Sigma, USA), was added. Three days later, the culture media was replaced with REGM for two days, and then changed into induction medium again. This sequence was repeated twice for a total 10 days. The inserts containing hypoxia-injured RTECs were renewed every three days. Following the immunofluorescence protocol described above, KSCs were stained with mouse anti-CK18 polyclonal antibody (Abcam, UK), mouse anti-E-cadherin polyclonal antibody, and rabbit anti-ZO-1 polyclonal antibody (Santa Cruz, USA). The induction strategy is shown in Fig 1.

Bottom Line: However, KSC proliferation, differentiation ability and tolerance to hypoxia were decreased in high-glucose cultures.Taken together, these results suggest the high-glucose microenvironment can damage the reparative ability of KSCs.It may result in a decreased of recovery capability of renal tubules from injury.

View Article: PubMed Central - PubMed

Affiliation: Department of Geriatric Nephrology, Chinese PLA General Hospital, State Key Laboratory of Kidney Disease, Beijing, China.

ABSTRACT
The incidence of acute kidney injury in patients with diabetes is significantly higher than that of patients without diabetes, and may be associated with the poor stemness capacity of kidney stem cells (KSCs) and limited recovery of injured renal tubules. To investigate the effects of hyperglycemic stress on KSC stemness, KSCs were isolated from the rat renal papilla and analyzed for their self-renewal and differentiation abilities. Our results showed that isolated KSCs expressed the mesenchymal stem cell markers N-cadherin, Nestin, CD133, CD29, CD90, and CD73. Moreover, KSCs co-cultured with hypoxia-injured renal tubular epithelial cell (RTECs) induced the expression of the mature epithelial cell marker CK18, suggesting that the KSCs could differentiate into RTECs in vitro. However, KSC proliferation, differentiation ability and tolerance to hypoxia were decreased in high-glucose cultures. Taken together, these results suggest the high-glucose microenvironment can damage the reparative ability of KSCs. It may result in a decreased of recovery capability of renal tubules from injury.

No MeSH data available.


Related in: MedlinePlus