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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

H&E Staining of murine pancreas tissue (A) cryosectioned to 20 microns before fixation. Staining was used to verify the tissue samples were intact, as subsequent sections were utilized for AFM stiffness measurements. Fluorescently tagged fibronectin gel (B) demonstrated that the fibronectin was homogeneously dispersed throughout the gel. A scratch can be seen in the image, which was necessary to provide a focusing point.
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Figure 1: H&E Staining of murine pancreas tissue (A) cryosectioned to 20 microns before fixation. Staining was used to verify the tissue samples were intact, as subsequent sections were utilized for AFM stiffness measurements. Fluorescently tagged fibronectin gel (B) demonstrated that the fibronectin was homogeneously dispersed throughout the gel. A scratch can be seen in the image, which was necessary to provide a focusing point.

Mentions: Measurements of unfixed native murine pancreas stiffness were taken on tissue samples cryosectioned and mounted on glass slides. The pancreas was sectioned to a thickness of 20 microns and the integrity was verified using H&E staining (Figure 1A). AFM force indentation measurements were then taken on n = 2 slide sections and approximately 50 force-indentation curves were taken on each at random locations. The Young’s modulus of elasticity for murine pancreas found was estimated as 1210 ± 77 Pa with individual measurements ranging from 618 Pa to 1407 Pa.


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)

H&E Staining of murine pancreas tissue (A) cryosectioned to 20 microns before fixation. Staining was used to verify the tissue samples were intact, as subsequent sections were utilized for AFM stiffness measurements. Fluorescently tagged fibronectin gel (B) demonstrated that the fibronectin was homogeneously dispersed throughout the gel. A scratch can be seen in the image, which was necessary to provide a focusing point.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: H&E Staining of murine pancreas tissue (A) cryosectioned to 20 microns before fixation. Staining was used to verify the tissue samples were intact, as subsequent sections were utilized for AFM stiffness measurements. Fluorescently tagged fibronectin gel (B) demonstrated that the fibronectin was homogeneously dispersed throughout the gel. A scratch can be seen in the image, which was necessary to provide a focusing point.
Mentions: Measurements of unfixed native murine pancreas stiffness were taken on tissue samples cryosectioned and mounted on glass slides. The pancreas was sectioned to a thickness of 20 microns and the integrity was verified using H&E staining (Figure 1A). AFM force indentation measurements were then taken on n = 2 slide sections and approximately 50 force-indentation curves were taken on each at random locations. The Young’s modulus of elasticity for murine pancreas found was estimated as 1210 ± 77 Pa with individual measurements ranging from 618 Pa to 1407 Pa.

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