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Early differentiation patterning of mouse embryonic stem cells in response to variations in alginate substrate stiffness.

Candiello J, Singh SS, Task K, Kumta PN, Banerjee I - J Biol Eng (2013)

Bottom Line: Upon generation of functionally mature ESC derived islet-like cells, they need to be implanted into diabetic patients to restore the loss of islet activity.Encapsulation in alginate microcapsules is a promising route of implantation, which can protect the cells from the recipient's immune system.The insight into these biophysical phenomena found in this study can be used along with other cues to enhance the differentiation of embryonic stem cells toward a specific lineage fate.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA. pkumta@pitt.edu.

ABSTRACT

Background: Embryonic stem cells (ESCs) have been implicated to have tremendous impact in regenerative therapeutics of various diseases, including Type 1 Diabetes. Upon generation of functionally mature ESC derived islet-like cells, they need to be implanted into diabetic patients to restore the loss of islet activity. Encapsulation in alginate microcapsules is a promising route of implantation, which can protect the cells from the recipient's immune system. While there has been a significant investigation into islet encapsulation over the past decade, the feasibility of encapsulation and differentiation of ESCs has been less explored. Research over the past few years has identified the cellular mechanical microenvironment to play a central role in phenotype commitment of stem cells. Therefore it will be important to design the encapsulation material to be supportive to cellular functionality and maturation.

Results: This work investigated the effect of stiffness of alginate substrate on initial differentiation and phenotype commitment of murine ESCs. ESCs grown on alginate substrates tuned to similar biomechanical properties of native pancreatic tissue elicited both an enhanced and incrementally responsive differentiation towards endodermal lineage traits.

Conclusions: The insight into these biophysical phenomena found in this study can be used along with other cues to enhance the differentiation of embryonic stem cells toward a specific lineage fate.

No MeSH data available.


Related in: MedlinePlus

Immunoflourescence Images of mESCs cultured on 10 mg/ml and 1× crosslinking alginate gels. Cells expressed high levels of CXCR4 (A) and AFP (B).
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Figure 5: Immunoflourescence Images of mESCs cultured on 10 mg/ml and 1× crosslinking alginate gels. Cells expressed high levels of CXCR4 (A) and AFP (B).

Mentions: For further verification of endoderm expression, we used immuno-flourescence imaging to verify protein expression of two of the higher upregulated genes found in the previous section. Immuno-staining of CXCR4 (Figure 5A) and AFP (Figure 5B) within a cell “clump” demonstrated clearly the presence of both proteins in the cells differentiated on the 10 mg/ml with 1X crosslinking alginate gel, selected as the representative gel condition.


Early differentiation patterning of mouse embryonic stem cells in response to variations in alginate substrate stiffness.

Candiello J, Singh SS, Task K, Kumta PN, Banerjee I - J Biol Eng (2013)

Immunoflourescence Images of mESCs cultured on 10 mg/ml and 1× crosslinking alginate gels. Cells expressed high levels of CXCR4 (A) and AFP (B).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3643844&req=5

Figure 5: Immunoflourescence Images of mESCs cultured on 10 mg/ml and 1× crosslinking alginate gels. Cells expressed high levels of CXCR4 (A) and AFP (B).
Mentions: For further verification of endoderm expression, we used immuno-flourescence imaging to verify protein expression of two of the higher upregulated genes found in the previous section. Immuno-staining of CXCR4 (Figure 5A) and AFP (Figure 5B) within a cell “clump” demonstrated clearly the presence of both proteins in the cells differentiated on the 10 mg/ml with 1X crosslinking alginate gel, selected as the representative gel condition.

Bottom Line: Upon generation of functionally mature ESC derived islet-like cells, they need to be implanted into diabetic patients to restore the loss of islet activity.Encapsulation in alginate microcapsules is a promising route of implantation, which can protect the cells from the recipient's immune system.The insight into these biophysical phenomena found in this study can be used along with other cues to enhance the differentiation of embryonic stem cells toward a specific lineage fate.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA. pkumta@pitt.edu.

ABSTRACT

Background: Embryonic stem cells (ESCs) have been implicated to have tremendous impact in regenerative therapeutics of various diseases, including Type 1 Diabetes. Upon generation of functionally mature ESC derived islet-like cells, they need to be implanted into diabetic patients to restore the loss of islet activity. Encapsulation in alginate microcapsules is a promising route of implantation, which can protect the cells from the recipient's immune system. While there has been a significant investigation into islet encapsulation over the past decade, the feasibility of encapsulation and differentiation of ESCs has been less explored. Research over the past few years has identified the cellular mechanical microenvironment to play a central role in phenotype commitment of stem cells. Therefore it will be important to design the encapsulation material to be supportive to cellular functionality and maturation.

Results: This work investigated the effect of stiffness of alginate substrate on initial differentiation and phenotype commitment of murine ESCs. ESCs grown on alginate substrates tuned to similar biomechanical properties of native pancreatic tissue elicited both an enhanced and incrementally responsive differentiation towards endodermal lineage traits.

Conclusions: The insight into these biophysical phenomena found in this study can be used along with other cues to enhance the differentiation of embryonic stem cells toward a specific lineage fate.

No MeSH data available.


Related in: MedlinePlus