P19081: Cell Factory
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Subsystem Build & Test

Table of Contents

Team Vision for Subsystem Level Build & Test Phase

During this phase, our team intended to complete all cell culture testing to obtain growth curve data that would determine the doubling time at various seeding densities. Along side with cell culture testing, we intended on beginning iterative fabrication process of the automation device. By the end of this phase, we were effectively able to acquire the doubling time for this cell type at various seeding densities as well as completing our initial build of the automation device.

Test Results Summary

Cell culture testing was performed in three 24-well plates over the course of eight days. Each plate was seeded as pictured to the right. Fluorescent imaging and hemocytometer cell counts were taken daily sampling from randomized wells. The media in each tray was replaced on day four of testing. Protocol for testing followed CELL-TM-01.

24-well Plate Layout

24-well Plate Layout

Testing on plate 1 was begun on January 23rd. The plate was seeded as shown above and there was a media swap that took place on day 4 of testing. The growth curve developed from plate 1 follows very closely to the expected shape of growth curve for these cells.

Plate 1 3000cells/cm2

Plate 1 3000cells/cm2

Plate 1 6000cells/cm2

Plate 1 6000cells/cm2

Plate 1 9000cells/cm2

Plate 1 9000cells/cm2

Plate 1 Growth Curve

Plate 1 Growth Curve

Testing on plate 2 was begun on January 24rd. The plate was seeded as shown above and there was a media swap that took place on day 4 of testing. The growth curve developed from plate 2 follows very closely to the expected shape of growth curve for these cells.

Plate 2 3000cells/cm2

Plate 2 3000cells/cm2

Plate 2 6000cells/cm2

Plate 2 6000cells/cm2

Plate 2 9000cells/cm2

Plate 2 9000cells/cm2

Plate 2 Growth Curve

Plate 2 Growth Curve

Testing on plate 3 was begun on January 30th. The plate was seeded as shown above and there was a media swap that took place on day 4 of testing. The growth curve developed from plate 3 does not follow the expected curve for this cell type. We determined to keep this data out of the doubling time calculation since it did not follow the expected trend.

Plate 3 3000cells/cm2

Plate 3 3000cells/cm2

Plate 3 6000cells/cm2

Plate 3 6000cells/cm2

Plate 3 9000cells/cm2

Plate 3 9000cells/cm2

Plate 3 Growth Curve

Plate 3 Growth Curve

The doubling time for cell seeding densities of 3000, 6000, and 9000 cells/cm2 were determined using the results from plates 1 and 2. These were determined using a linear trend line of the data points between day 1 and day 5.

Doubling Time at Varying Cell Seeding Densities

Doubling Time at Varying Cell Seeding Densities

Live Document:Test Plan

Live Document:Test Results

Risk and Problem Tracking

After our cell culture testing, both our risk management and problem tracking documents were updated to reflect new risks or problems encountered. Our risk management added the school closing and not having lab access, not enough ports available on the micro controller, and a team member not being able to make it in for testing due to travel restrictions. The problem tracking document follows the resolution of these problems.
Problem Tracking

Problem Tracking

Live Document:Problem Tracking
Risk Management

Risk Management

Live Document:Risk Management

Automation Summary

Automated Passaging Machine

Automated Passaging Machine

Shown in the figure above is the first build of the Automated Cell Passaging System. We have begun integration of the liquid removal and liquid dispensing subsystems and have completed an estimated 75% of the mechanical build. We have not conducted any official testing of any of the subsystems, however, we have successfully programmed the 2-axis stepper motor system and have also successfully dispensed liquids in accurate volumes into a petri dish on our previous test fixture.

Dispensing Subsystem

Dispensing Subsystem

The dispensing subsystem is intended to be capable of dispensing precise volumes of growth media, PBS, and trypsin on demand as per the cell passaging process. We still have to determine the appropriate way to create a pressure seal with the GL45 screw caps for aseptic liquid transfer within our budget limitations. The previous item we were going to order for this purpose has been discontinued by one of our suppliers, Cole-Palmer.

Wiring Diagram

Wiring Diagram

We have encountered an issue with our control system. The problem is that there are a total of 4 12VDC devices that we need to control independently (3 liquid valves, and 1 pneumatic valve). There are only 2 available 12V ports on the microcontroller. The wiring diagram above shows a possible solution to this issue using solid state relays that are controlled by the other two available motor ports.

Bill of Materials

Bill of Materials

Live Document:Bill of Materials

Functional Demo Materials

Include links to:

Plans for next phase

By the end of next phase, our team intends to have a large portion of the LabVIEW coding completed to implement the doubling time results. We also intend to complete baseline testing for the automation portion of the project. A key focus will be on integrating the halves of the project.

Tony Yosick's Three Week Plan Live Document: Tony's Goals

Philip Tinder's Three Week Plan Live Document: Philip's Goals

Christa Vuglar's Three Week Plan Live Document: Christa' Goals

Charles Henle's Three Week Plan Live Document: Charles' Goals

Emma Kate Flanagan's Three Week Plan Live Document: Emma Kate's Goals


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