External Deliverables

  • This is my reference guide for basic Solidworks Assembly Features.

  • This is the abstract I helped write before submitting to RMBS.

  • This is a short overview paper of what I have researched on cell deposition.

Internal Deliverables

  • Pictured are cells creating spherical structures in the test of our Agarose molds.

  • In this picture, one can see the mold and printhead fitting in our fixture.

  • This link pictured will take you to my video of flow using our 3D printed Fixture.

Culture Experiment

I started an experiment with our agarose wells. I took a 90% confluent and an 80% confluent osteoblast cell culture through the splitting process. The cells were distributed into a 10 mL media solution before being split into 5 mL tubes and spun in the centrifuge for 6 minutes at 1000 rpm. Then, the media was piped out and replaced with new media which was used to flush cells off the walls and break up any cell clumps. Then, approx .25 mL of this media was piped into each Agarose mold. The cultures were split into different petri dishes depending on the parent culture. The petri dishes were then sealed and placed in an incubator at 37 degrees C and 5% Carbon Dioxide. Culture was checked at 3 days and pictures taken at 5. Culture slice and dead cell stain done later due to time constraints. Culture media determined to needing replacing daily which is difficult without sucking up the cell cultures too. This is where the microfluidic device would come into play with replacing cell media through a PDMS printhead which could keep Oxygen and Carbon Dioxide levels adequate due to being highly permeable to those gases.

Where to Go Next Semester

I have a few things where I want to accomplish early next semester as some next steps. I want to perform an experiment where I would pipe 40% confluent cells into the agarose gel molds like I did previously. Instead of leaving them and trying to pipe out and in media every day, I would pipe out liquid the next day and stick in the PDMS printhead and start flowing media to it in order to keep it alive. If the cells don't get sucked up or impede a good seal, it would feasibly not let the cells flow back up through the waste tube with the media. This would circumvent the problem of cells not sticking together in the wells and just flowing out through the media.

The second place to go involving cells next semester would be tracking cells through the printhead via our microPIV experiments. These are explained in our research abstract. I may actually gear some work towards understanding and performing microPIV experiments and simulations with that part of our group. However, one experiment we want to perform is infusing cells with red or green fluorescence. This would allow our laser to be able to track individual cells or aggregates through the printhead.