April 2023:
Matched project and gathered teammates
Met Dr. Anderson and discussed goals/scope of project
Did lab safety training and took lab tour to familiarize ourselves with wet lab
Processed first round of porcine hearts
May 2023:
Made first round of decellularization solution (1% SDS)
Attempted first perfusion and immersion decellularization attempts
Planned out project in more detail
Discussed how ECM from different chambers will have different cardiomyocyte phenotypes and behavior
Researched ways to test mechanical properties of hydrogels
June 2023:
Finalized adapting Christman lab decell protocol for our purposes
Performed first attempt at Christman lab decell protocol on processed hearts
July 2023:
Continued performing decellularization protocol to build up stock of decellularized ECM (dECM)
Did preliminary histology on dECM stock
August 2023:
Made adjustments based on previous runs and performed Christman decell protocol again
Lasercut clips and frames for EHT testing
Went to Christman lab and milled samples with 40 grade filter
Performed first attempt at pepsin digestion to produce hydrogel
September 2023:
Discussed how to use the 3d printed grips made in August
October 2023:
Planned out goals and experiments for the quarter and year
Planned meeting times for experiments, journal club, and milling at Christman lab
Performed second and third attempts at pepsin digestion protocol
Performed more decellularizations to build stock
Met with whole group to analyze why we haven't been achieving gelation and plan next steps for experiments
November 2023:
Went over all existing protocols and updated them to fix issues/feedback
Scheduled experiments for upcoming months
Did first large scale decell
December 2023:
Discussed which experiments we want to run to finish the quarter
Performed histology on decell batch 5 samples
Wrote, edited, and finalized senior design proposal
January 2024:
Regrouped after break to discuss goals for the year and plan experiments
Section heart tissue into slices for new one day decell protocol
Ran first timepoint study by removing tissue and supernatant at regular intervals
Ran decell using SDC vs SDS to compare
February 2024:
Ran pepsin digestion protocol while varying salinity and ion presence to test if these were the causes of our previous gelation issues
Some visual gelling at first, but not after incubation
Performed DC protein assay to test protein concentrations
Ran pepsin digestion protocol over ice, and achieved gelation
Ran TRIzol DNA isolation to check effectiveness of decell
March 2024:
Performed more DNA quantification to test effectiveness of SDC vs SDS protocols
Performed protein release assays to test effectiveness of SDC vs SDS protocols
Wrote and edited senior design final report
April 2024:
Optimized shavings decell protocol
Ran multiple decells and timepoint studies
Performed protein assay and DNA quantification on samples
May 2024:
Wrapped up experiments and organized project
Prepared poster and gave final presentation
Finalized senior design final report
Presented at Bioengineering day
Rohil Deolalikar