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Culturing DRRAGN cardiomyocytes on GelTA and GelTA-MWCNT to assess differences in maturity level

Stoter, A.M.S. (2019) Culturing DRRAGN cardiomyocytes on GelTA and GelTA-MWCNT to assess differences in maturity level.

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Abstract:Background: A model of in vitro cultured mature cardiomyocytes is essential to fabricate disease models and research drug responses. Over the past years, research has been done into the use of gelatin based hydrogel scaffolds for the culturing of cardiomyocytes. However, pristine gelatin based hydrogel scaffolds lack conductivity, which is present in human cardiac tissue. Here, a method to fabricate a conductive gelatin based hydrogel scaffold is discussed. Method: Gelatin tyramine has been mixed with multi-walled carbon nanotubes to achieve a higher conductivity of the hydrogel. To investigate the effect of the multi-walled carbon nanotubes on the conductivity of the hydrogel, a conductivity measurement is done with a four point probe. To investigate the effect of the multi-walled carbon nanotubes on the maturity of the cardiomyocytes, the RNA expression of the gene encoding connexin 43 is measured with qPCR and the expression of the connexin 43 protein is imaged with immunostaining. The orientation of �-actinine strands has also been investigated. Results and discussion: It has been found that the presence of multi-walled carbon nanotubes did not show a positive correlation to the conductivity of the scaffolds. It is expected that this is due to the lack of a network formed by the multi-walled carbon nanotubes, causing the formation of electrical double layers at the interfaces between the carbon nanotubes and the gel. With regard to the effect of the presence of carbon nanotubes on the maturity of the cultured cardiomyocytes it is hard to draw a concrete conclusion due to the fashion in which the cardiomyocytes are cultured, as multiple cardiomyocytes are present on top of each other. Conclusion and recommendations: During the scope of this master thesis, a design for the fabrication of thin gelatine based hydrogel layers has been optimized. For further optimization it is recommended to apply an electric field over the hydrogels with carbon nanotubes during the curing of the hydrogels, to allow the carbon nanotubes to align. This would form a network of carbon nanotubes in the gel, causing less electrical double layers to be formed. For the fabrication of the thin hydrogel layers it is recommended to use a PMMA mold for reproducibility.
Item Type:Essay (Master)
Faculty:TNW: Science and Technology
Subject:42 biology
Programme:Biomedical Engineering MSc (66226)
Link to this item:http://purl.utwente.nl/essays/79165
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