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Differentiation of Wharton's jelly mesenchymal stem cells into neurons in alginate scaffold.

Hosseini SM, Vasaghi A, Nakhlparvar N, Roshanravan R, Talaei-Khozani T, Razi Z - Neural Regen Res (2015)

Bottom Line: In this study, we cultured WJMSCs in a three-dimensional scaffold fabricated by 0.25% alginate and 50 mM CaCl2 in the presence of neurogenic medium containing 10 μM retinoic acid and 20 ng/mL basic fibroblast growth factor.After 10 days, immunofluorescence staining was performed for detecting β-tubulin (marker for WJMSCs-differentiated neuron) and CD271 (motor neuron marker). β-Tubulin and CD271 expression levels were significantly greater in the WJMSCs cultured in the three-dimensional alginate scaffold than in the conventional two-dimensional culture condition.These findings suggest that three-dimensional alginate scaffold cell culture system can induce neuronal differentiation of WJMSCs effectively.

View Article: PubMed Central - PubMed

Affiliation: Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran ; Cell and Molecular Medicine Student Research Group, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran ; Stem Cell Laboratory, Department of Anatomy, Shiraz University of Medical Sciences, Shiraz, Iran.

ABSTRACT
Alginate scaffold has been considered as an appropriate biomaterial for promoting the differentiation of embryonic stem cells toward neuronal cell lineage. We hypothesized that alginate scaffold is suitable for culturing Wharton's jelly mesenchymal stem cells (WJMSCs) and can promote the differentiation of WJMSCs into neuron-like cells. In this study, we cultured WJMSCs in a three-dimensional scaffold fabricated by 0.25% alginate and 50 mM CaCl2 in the presence of neurogenic medium containing 10 μM retinoic acid and 20 ng/mL basic fibroblast growth factor. These cells were also cultured in conventional two-dimensional culture condition in the presence of neurogenic medium as controls. After 10 days, immunofluorescence staining was performed for detecting β-tubulin (marker for WJMSCs-differentiated neuron) and CD271 (motor neuron marker). β-Tubulin and CD271 expression levels were significantly greater in the WJMSCs cultured in the three-dimensional alginate scaffold than in the conventional two-dimensional culture condition. These findings suggest that three-dimensional alginate scaffold cell culture system can induce neuronal differentiation of WJMSCs effectively.

No MeSH data available.


Wharton's jelly mesenchymal stem cells (WJMSCs) had the capability to differentiate into adipocytes (A) and osteocytes (B).Arrows show the nuclei of the cells and star shows calcium deposit. WJMSCs could be stained with oil red S (A) and alizarin red S (B). Scale bars: 50 μm.
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Figure 2: Wharton's jelly mesenchymal stem cells (WJMSCs) had the capability to differentiate into adipocytes (A) and osteocytes (B).Arrows show the nuclei of the cells and star shows calcium deposit. WJMSCs could be stained with oil red S (A) and alizarin red S (B). Scale bars: 50 μm.

Mentions: The cells were shown to be positive for CD44 (88.87%), CD105 (5.75%), CD90 (82.87%) and CD73 (97.08%). The percentages of the cells positive for CD144 (0.04%) and CD34 (1.07%) were negligible (Figure 1). Alizarin red S and oil red O staining also revealed that the cells were capable to differentiate toward osteoblasts and adipocytes, respectively (Figure 2).


Differentiation of Wharton's jelly mesenchymal stem cells into neurons in alginate scaffold.

Hosseini SM, Vasaghi A, Nakhlparvar N, Roshanravan R, Talaei-Khozani T, Razi Z - Neural Regen Res (2015)

Wharton's jelly mesenchymal stem cells (WJMSCs) had the capability to differentiate into adipocytes (A) and osteocytes (B).Arrows show the nuclei of the cells and star shows calcium deposit. WJMSCs could be stained with oil red S (A) and alizarin red S (B). Scale bars: 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4590246&req=5

Figure 2: Wharton's jelly mesenchymal stem cells (WJMSCs) had the capability to differentiate into adipocytes (A) and osteocytes (B).Arrows show the nuclei of the cells and star shows calcium deposit. WJMSCs could be stained with oil red S (A) and alizarin red S (B). Scale bars: 50 μm.
Mentions: The cells were shown to be positive for CD44 (88.87%), CD105 (5.75%), CD90 (82.87%) and CD73 (97.08%). The percentages of the cells positive for CD144 (0.04%) and CD34 (1.07%) were negligible (Figure 1). Alizarin red S and oil red O staining also revealed that the cells were capable to differentiate toward osteoblasts and adipocytes, respectively (Figure 2).

Bottom Line: In this study, we cultured WJMSCs in a three-dimensional scaffold fabricated by 0.25% alginate and 50 mM CaCl2 in the presence of neurogenic medium containing 10 μM retinoic acid and 20 ng/mL basic fibroblast growth factor.After 10 days, immunofluorescence staining was performed for detecting β-tubulin (marker for WJMSCs-differentiated neuron) and CD271 (motor neuron marker). β-Tubulin and CD271 expression levels were significantly greater in the WJMSCs cultured in the three-dimensional alginate scaffold than in the conventional two-dimensional culture condition.These findings suggest that three-dimensional alginate scaffold cell culture system can induce neuronal differentiation of WJMSCs effectively.

View Article: PubMed Central - PubMed

Affiliation: Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran ; Cell and Molecular Medicine Student Research Group, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran ; Stem Cell Laboratory, Department of Anatomy, Shiraz University of Medical Sciences, Shiraz, Iran.

ABSTRACT
Alginate scaffold has been considered as an appropriate biomaterial for promoting the differentiation of embryonic stem cells toward neuronal cell lineage. We hypothesized that alginate scaffold is suitable for culturing Wharton's jelly mesenchymal stem cells (WJMSCs) and can promote the differentiation of WJMSCs into neuron-like cells. In this study, we cultured WJMSCs in a three-dimensional scaffold fabricated by 0.25% alginate and 50 mM CaCl2 in the presence of neurogenic medium containing 10 μM retinoic acid and 20 ng/mL basic fibroblast growth factor. These cells were also cultured in conventional two-dimensional culture condition in the presence of neurogenic medium as controls. After 10 days, immunofluorescence staining was performed for detecting β-tubulin (marker for WJMSCs-differentiated neuron) and CD271 (motor neuron marker). β-Tubulin and CD271 expression levels were significantly greater in the WJMSCs cultured in the three-dimensional alginate scaffold than in the conventional two-dimensional culture condition. These findings suggest that three-dimensional alginate scaffold cell culture system can induce neuronal differentiation of WJMSCs effectively.

No MeSH data available.