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Three-Dimensional Aggregates Enhance the Therapeutic Effects of Adipose Mesenchymal Stem Cells for Ischemia-Reperfusion Induced Kidney Injury in Rats.

Zhao X, Qiu X, Zhang Y, Zhang S, Gu X, Guo H - Stem Cells Int (2015)

Bottom Line: Low engraftment of stem cells, however, limits the therapeutic effects of AdMSCs.In vitro data indicated that AdMSCs in 3D aggregates were less susceptible to oxidative and hypoxia stress induced by 200 μM peroxide and hypoxia/reoxygenation, respectively, compared with those cultured in two-dimensional (2D) monolayer.Improved grafted AdMSCs were observed in kidney injected with 3D aggregates compared with AdMSCs cultured in 2D monolayer.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Affiliated Drum Tower Hospital, School of Medicine, Nanjing University, Nanjing 210008, China ; Institute of Urology, Nanjing University, Nanjing 210093, China.

ABSTRACT
It has been shown that administration of adipose derived mesenchymal stem cells (AdMSCs) enhanced structural and functional recovery of renal ischemia-reperfusion (IR) injury. Low engraftment of stem cells, however, limits the therapeutic effects of AdMSCs. The present study was designed to enhance the therapeutic effects of AdMSCs by delivering AdMSCs in a three-dimensional (3D) aggregates form. Microwell was used to produce 3D AdMSCs aggregates. In vitro data indicated that AdMSCs in 3D aggregates were less susceptible to oxidative and hypoxia stress induced by 200 μM peroxide and hypoxia/reoxygenation, respectively, compared with those cultured in two-dimensional (2D) monolayer. Furthermore, AdMSCs in 3D aggregates secreted more proangiogenic factors than those cultured in 2D monolayer. 2D AdMSCs or 3D AdMSCs aggregates were injected into renal cortex immediately after induction of renal IR injury. In vivo data revealed that 3D aggregates enhanced the effects of AdMSCs in recovering function and structure after renal IR injury. Improved grafted AdMSCs were observed in kidney injected with 3D aggregates compared with AdMSCs cultured in 2D monolayer. Our results demonstrated that 3D AdMSCs aggregated produced by microwell enhanced the retention and therapeutic effects of AdMSCs for renal IR injury.

No MeSH data available.


Related in: MedlinePlus

Characters of 3D AdMSCs aggregates. (a) Viability of AdMSCs cultured in 2D monolayer. (b) Viability of 3D AdMSCs aggregates 1 d, 3 d, and 7 d after seeding in microwell. Live or dead cells were stained with FDA (live, green) PI (dead, red). (c) Fluorescent images of 2D AdMSCs or 3D aggregates stained with fibronectin and laminin. (d) mRNA expression of fibronectin and laminin in 2D AdMSCs or 3D aggregates. ∗P < 0.05 compared with 2D AdMSCs.
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fig2: Characters of 3D AdMSCs aggregates. (a) Viability of AdMSCs cultured in 2D monolayer. (b) Viability of 3D AdMSCs aggregates 1 d, 3 d, and 7 d after seeding in microwell. Live or dead cells were stained with FDA (live, green) PI (dead, red). (c) Fluorescent images of 2D AdMSCs or 3D aggregates stained with fibronectin and laminin. (d) mRNA expression of fibronectin and laminin in 2D AdMSCs or 3D aggregates. ∗P < 0.05 compared with 2D AdMSCs.

Mentions: As shown in Figure 2(b), most cells in 3D aggregates were positive for FDA and negative for PI staining, suggesting high viability in the aggregates for at least 7 days (Figures 2(a) and 2(b)). 3D AdMSCs aggregates showed well-preserved ECM compared to the 2D AdMSCs. After culture in microwell for 7 days, cells expressed significantly increased the level of fibronectin and laminin (Figures 2(c)–2(e)) compared with 2D cells, partly indicating the significantly increased secretion of ECM in 3D AdMSCs aggregates.


Three-Dimensional Aggregates Enhance the Therapeutic Effects of Adipose Mesenchymal Stem Cells for Ischemia-Reperfusion Induced Kidney Injury in Rats.

Zhao X, Qiu X, Zhang Y, Zhang S, Gu X, Guo H - Stem Cells Int (2015)

Characters of 3D AdMSCs aggregates. (a) Viability of AdMSCs cultured in 2D monolayer. (b) Viability of 3D AdMSCs aggregates 1 d, 3 d, and 7 d after seeding in microwell. Live or dead cells were stained with FDA (live, green) PI (dead, red). (c) Fluorescent images of 2D AdMSCs or 3D aggregates stained with fibronectin and laminin. (d) mRNA expression of fibronectin and laminin in 2D AdMSCs or 3D aggregates. ∗P < 0.05 compared with 2D AdMSCs.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4663369&req=5

fig2: Characters of 3D AdMSCs aggregates. (a) Viability of AdMSCs cultured in 2D monolayer. (b) Viability of 3D AdMSCs aggregates 1 d, 3 d, and 7 d after seeding in microwell. Live or dead cells were stained with FDA (live, green) PI (dead, red). (c) Fluorescent images of 2D AdMSCs or 3D aggregates stained with fibronectin and laminin. (d) mRNA expression of fibronectin and laminin in 2D AdMSCs or 3D aggregates. ∗P < 0.05 compared with 2D AdMSCs.
Mentions: As shown in Figure 2(b), most cells in 3D aggregates were positive for FDA and negative for PI staining, suggesting high viability in the aggregates for at least 7 days (Figures 2(a) and 2(b)). 3D AdMSCs aggregates showed well-preserved ECM compared to the 2D AdMSCs. After culture in microwell for 7 days, cells expressed significantly increased the level of fibronectin and laminin (Figures 2(c)–2(e)) compared with 2D cells, partly indicating the significantly increased secretion of ECM in 3D AdMSCs aggregates.

Bottom Line: Low engraftment of stem cells, however, limits the therapeutic effects of AdMSCs.In vitro data indicated that AdMSCs in 3D aggregates were less susceptible to oxidative and hypoxia stress induced by 200 μM peroxide and hypoxia/reoxygenation, respectively, compared with those cultured in two-dimensional (2D) monolayer.Improved grafted AdMSCs were observed in kidney injected with 3D aggregates compared with AdMSCs cultured in 2D monolayer.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Affiliated Drum Tower Hospital, School of Medicine, Nanjing University, Nanjing 210008, China ; Institute of Urology, Nanjing University, Nanjing 210093, China.

ABSTRACT
It has been shown that administration of adipose derived mesenchymal stem cells (AdMSCs) enhanced structural and functional recovery of renal ischemia-reperfusion (IR) injury. Low engraftment of stem cells, however, limits the therapeutic effects of AdMSCs. The present study was designed to enhance the therapeutic effects of AdMSCs by delivering AdMSCs in a three-dimensional (3D) aggregates form. Microwell was used to produce 3D AdMSCs aggregates. In vitro data indicated that AdMSCs in 3D aggregates were less susceptible to oxidative and hypoxia stress induced by 200 μM peroxide and hypoxia/reoxygenation, respectively, compared with those cultured in two-dimensional (2D) monolayer. Furthermore, AdMSCs in 3D aggregates secreted more proangiogenic factors than those cultured in 2D monolayer. 2D AdMSCs or 3D AdMSCs aggregates were injected into renal cortex immediately after induction of renal IR injury. In vivo data revealed that 3D aggregates enhanced the effects of AdMSCs in recovering function and structure after renal IR injury. Improved grafted AdMSCs were observed in kidney injected with 3D aggregates compared with AdMSCs cultured in 2D monolayer. Our results demonstrated that 3D AdMSCs aggregated produced by microwell enhanced the retention and therapeutic effects of AdMSCs for renal IR injury.

No MeSH data available.


Related in: MedlinePlus