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In developing and optimizing the hydrogel protocols, there were many technical aspects and solutions considered. A variety of laboratory techniques were employed to decellularize the tissue and assess for DNA and protein content. 3D printing was also employed to design tools for future testing of the mechanical properties.
Comparison of SDS and SDC 8 day protocol adapted from Dr. Karen Christmans lab. The SDC was not able to fully decellularize in this time frame and further optimization was required.
Optimized timepoint decellularization study. This 5 hour protocol illustrates how tissue samples and supernatant can be tested to determine DC protein release and decellularization efficiency at any point in the study. We were able to show that both SDS and SDC methods were able to get to the decellularization standard of 50ng/ul of DNA content after decell and over time SDC shows to have a gentler decell which can be concluded from a decreased amount of released protein content in the supernatant overtime. In the future, that may prove that for the construction of EHT's specifically SDC is a better detergent that is capable of preserving more structural proteins and bioactive molecules to make more accurate in vitro EHT models.
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