Bioreactor for Tissue Engineering, Team Bioreactor Week 10 & 11 – November 16 to November 29, 2007 Team Members: Rachel Mosher – Team leader Kara Barnhart – Communications, BSAC Joel Gaston – BWIG Client: Dr. Susan Thibeault Dept. of Otolaryngology, UW Medical School Phone: 608/263-6751, Email: [email protected] Advisor: Brenda Ogle Assistant Professor/ Dept. of Biomedical Engineering Phone: 608/265-8267, Email: [email protected] Problem Statement The aim of this project is to re-design and improve upon a previous version of a bioreactor that will be used for the culturing of human vocal fold fibroblasts. The previous design was able to vibrate two pairs of cell-seeded strips under tensile stress, but had design flaws that needed improvement, including keeping the bioreactor leak-proof, subjecting the cells to more stimuli, and allowing the equipment providing the stimuli to be controlled by a computer. Our goals are to finish the design and fabrication of this new model, to obtain a substitute for the cellular substrate, Tecoflex, and to test the bioreactor and cellular substrate for optimal design and operating conditions. The bioreactor will be made from two T-flasks, two moving magnet linear voice coil actuators, two rotary stepper motors, and two linear stepper motors. A total of four pairs of cells will be immersed in a buffer in two T-flasks, and subjected to vibration, tensile stress, and angular changes between each pair of strips. This device needs to be easily replaced with disposable and/or sterilizable parts, fit inside a standard incubator, and have a capacity to generate vibrations within the frequency range of 50-400 Hz. Restatement of Team Goals The team will meet on Wednesday, November 28 to discuss progress with putting the bioreactor together, cell culture, and the previous week’s testing of the cellular substrates. The team will meet with the client on Friday, November 16 at SMI to update the client on the team’s progress. Individual Goals Joel Update website, work on cell culture, determine testing protocols, Start work on poster Kara Continue making Tecoflex and hydrogels, start testingRachel Machine bioreactor parts, work on next week’s progress report. Summary of Accomplishment The team met on Wednesday, November 28 to discuss cellular substrate progress, the poster, and testing plans for next week. During class time on Friday, November 16, the team discussed each member’s progress, met with Professor Ogle, and met with the client. Statement of Team Goals The team will finish up preparing/machining the bioreactor for testing, buying the remainder of parts needed, preparing and testing the cell culture materials, and working on the poster and presentation. Difficulties There has been difficulty getting into SMI to make cellular substrates and some difficulty finding proper springs, screws and aluminum stock. Activities 11.15.2007 Went to the lab and continued making the Tecoflex, started another batch of Tecoflex 1.50h 11.16.2007 Continued making the Tecoflex from the day before and started a new batch 2.00h 11.19.2007 Put one batch of the Tecoflex on distilled water, put other batch in a mold, talked to Stephanie 1.00h 11.20.2007 Lyophilized one batch of Tecoflex, started a new batch of Tecoflex 2.00h 11.21.2007 Took Tecoflex out of the vacuum, checked on Tecoflex started the day before 0.75h 11.26.2007 Put Tecoflex on distilled water, worked with the old batches of Tecoflex 0.75h 11.27.2007 Lyophilized Tecoflex, made hydrogels 2.50h Kara 11.28.2007 Observed hydrogel, worked with older batches of Tecoflex 1.00h 11.50h 11.15.2007 Went to lab, cultured and split cells, talked to Stephanie about seeding strips with cells. Finished suspension system, including knots and testing 2.25h Joel 11.16.2007 Worked on poster, gathering information and determining layout 1.5h 10.25h11.17.2007 Worked on poster, adding information 2h 11.19.2007 Went to lab, cultured cells, continued research into force transducers 1.5h 11.21.2007 Went to lab, cultured and split cells .75h 11.26.2007 Went to lab, cultured cells. Worked on poster, adding information 1.5h 11.29.2007 Researched places to buy springs with our dimensions 1.25h 11.25.2007 Machined holders for rotary stepper motors, started machining threaded rods to attach to stepper motors, machined support holders for end of threaded rods opposite motors. 6.25h Rachel 11.29.2007 Searched on-line for spring vendors, went to Ace Hardware on Midvale Blvd to find screws and aluminum stock, completed and e-mailed Progress Report for week 10. 1.00h 7.25h Team 11.28.2007 Met outside of class to discuss team members’ progress, finish and test suspension system, and made future plans for testing. 1.75h 1.75h Project Timeline September October November December Tasks 17 24 1 8 15 22 29 5 12 19 26 3 10 Project research Brainstorming PDS Prototype design Prototype building Actual device design Ordering Expected shipping Device manufacturingTesting Re-designing Re-testing Meeting with client Group meeting Presentation Progress report Website Expenses The expenses currently total ~$150.Testing protocols Day 1: Testing the substrate in T-flask Purpose: To determine how the substrates behave in the media needed to grow the cells, including separation of substrate from Tecoflex, and change in pH of media Equipment: Four clean, dry T-flasks, 60mL cell media, both substrates to be tested Notes: If the color of the media changes to yellow, it is too acidic. If the color of the media changes to red, it is too basic. Procedure: Testing first substrate: First trial: 10mL of media • Obtain a clean T-flask and cell media • Place the two strips to be tested in the T-flask • Add 10mL of media to flask • Let sit for 30 minutes • Record results, including any separation of Tecoflex and substrate, and color change
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