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Controlled electromechanical cell stimulation on-a-chip.

Pavesi A, Adriani G, Rasponi M, Zervantonakis IK, Fiore GB, Kamm RD - Sci Rep (2015)

Bottom Line: The platform was validated in experiments using human bone marrow mesenchymal stem cells.These experiments demonstrated the ability for inducing changes in cell morphology, cytoskeletal fiber orientation and changes in gene expression under physiological stimuli.This novel bioengineering approach can be readily applied to various studies, especially in the fields of stem cell biology and regenerative medicine.

View Article: PubMed Central - PubMed

Affiliation: Biosym IRG, Singapore-MIT Alliance for Research and Technology, Singapore.

ABSTRACT
Stem cell research has yielded promising advances in regenerative medicine, but standard assays generally lack the ability to combine different cell stimulations with rapid sample processing and precise fluid control. In this work, we describe the design and fabrication of a micro-scale cell stimulator capable of simultaneously providing mechanical, electrical, and biochemical stimulation, and subsequently extracting detailed morphological and gene-expression analysis on the cellular response. This micro-device offers the opportunity to overcome previous limitations and recreate critical elements of the in vivo microenvironment in order to investigate cellular responses to three different stimulations. The platform was validated in experiments using human bone marrow mesenchymal stem cells. These experiments demonstrated the ability for inducing changes in cell morphology, cytoskeletal fiber orientation and changes in gene expression under physiological stimuli. This novel bioengineering approach can be readily applied to various studies, especially in the fields of stem cell biology and regenerative medicine.

No MeSH data available.


Related in: MedlinePlus

Immunofluorescence data.(A) Actin cytoskeleton, nuclei, and CX43 expression in control and electromechanically stimulated devices at 3% strain and 5 V/cm electrical stimulation. Scale bar is 100 μm. (B) Quantification and statistical analysis of the CX43 fluorescence intensity under three different conditions: control, 3% strain with 5 V/cm electrical stimulation, and 7% strain with 5 V/cm electrical stimulation. Data are expressed as means ± SD. Two-tailed t-test, *p < 0.05. (C) Cell density after 14 days of culture in control and stimulated devices (3% strain and 5 V/cm). Data are expressed as means ± SD.
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f5: Immunofluorescence data.(A) Actin cytoskeleton, nuclei, and CX43 expression in control and electromechanically stimulated devices at 3% strain and 5 V/cm electrical stimulation. Scale bar is 100 μm. (B) Quantification and statistical analysis of the CX43 fluorescence intensity under three different conditions: control, 3% strain with 5 V/cm electrical stimulation, and 7% strain with 5 V/cm electrical stimulation. Data are expressed as means ± SD. Two-tailed t-test, *p < 0.05. (C) Cell density after 14 days of culture in control and stimulated devices (3% strain and 5 V/cm). Data are expressed as means ± SD.

Mentions: In mature cardiac cells, the CX43 gap junction protein is essential for intercellular communication when single cells constitute a functional pluricellular complex. Therefore, CX43 serves as a marker of induced differentiation37, and its expression was evaluated here by fluorescence immunostaining (Fig. 5A). There were statistically significant differences in CX43 intensities between control samples and cells that underwent electrical stimulation (5 V/cm) coupled with low strain levels (3%) (Fig. 5B). No significant differences were observed between control cells and cells exposed to electromechanical stimulation with a higher strain level (7%). Moreover, no differences in cell density were observed between the stimulated (3% strain and 5 V/cm) and control samples after 14 days in culture (Fig. 5C).


Controlled electromechanical cell stimulation on-a-chip.

Pavesi A, Adriani G, Rasponi M, Zervantonakis IK, Fiore GB, Kamm RD - Sci Rep (2015)

Immunofluorescence data.(A) Actin cytoskeleton, nuclei, and CX43 expression in control and electromechanically stimulated devices at 3% strain and 5 V/cm electrical stimulation. Scale bar is 100 μm. (B) Quantification and statistical analysis of the CX43 fluorescence intensity under three different conditions: control, 3% strain with 5 V/cm electrical stimulation, and 7% strain with 5 V/cm electrical stimulation. Data are expressed as means ± SD. Two-tailed t-test, *p < 0.05. (C) Cell density after 14 days of culture in control and stimulated devices (3% strain and 5 V/cm). Data are expressed as means ± SD.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488866&req=5

f5: Immunofluorescence data.(A) Actin cytoskeleton, nuclei, and CX43 expression in control and electromechanically stimulated devices at 3% strain and 5 V/cm electrical stimulation. Scale bar is 100 μm. (B) Quantification and statistical analysis of the CX43 fluorescence intensity under three different conditions: control, 3% strain with 5 V/cm electrical stimulation, and 7% strain with 5 V/cm electrical stimulation. Data are expressed as means ± SD. Two-tailed t-test, *p < 0.05. (C) Cell density after 14 days of culture in control and stimulated devices (3% strain and 5 V/cm). Data are expressed as means ± SD.
Mentions: In mature cardiac cells, the CX43 gap junction protein is essential for intercellular communication when single cells constitute a functional pluricellular complex. Therefore, CX43 serves as a marker of induced differentiation37, and its expression was evaluated here by fluorescence immunostaining (Fig. 5A). There were statistically significant differences in CX43 intensities between control samples and cells that underwent electrical stimulation (5 V/cm) coupled with low strain levels (3%) (Fig. 5B). No significant differences were observed between control cells and cells exposed to electromechanical stimulation with a higher strain level (7%). Moreover, no differences in cell density were observed between the stimulated (3% strain and 5 V/cm) and control samples after 14 days in culture (Fig. 5C).

Bottom Line: The platform was validated in experiments using human bone marrow mesenchymal stem cells.These experiments demonstrated the ability for inducing changes in cell morphology, cytoskeletal fiber orientation and changes in gene expression under physiological stimuli.This novel bioengineering approach can be readily applied to various studies, especially in the fields of stem cell biology and regenerative medicine.

View Article: PubMed Central - PubMed

Affiliation: Biosym IRG, Singapore-MIT Alliance for Research and Technology, Singapore.

ABSTRACT
Stem cell research has yielded promising advances in regenerative medicine, but standard assays generally lack the ability to combine different cell stimulations with rapid sample processing and precise fluid control. In this work, we describe the design and fabrication of a micro-scale cell stimulator capable of simultaneously providing mechanical, electrical, and biochemical stimulation, and subsequently extracting detailed morphological and gene-expression analysis on the cellular response. This micro-device offers the opportunity to overcome previous limitations and recreate critical elements of the in vivo microenvironment in order to investigate cellular responses to three different stimulations. The platform was validated in experiments using human bone marrow mesenchymal stem cells. These experiments demonstrated the ability for inducing changes in cell morphology, cytoskeletal fiber orientation and changes in gene expression under physiological stimuli. This novel bioengineering approach can be readily applied to various studies, especially in the fields of stem cell biology and regenerative medicine.

No MeSH data available.


Related in: MedlinePlus