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Sterility Test
Sterility Test
The aim for this evaluation is to verify that the device will not get contaminated when in use. The protocols created in the previous section called “adherent cell process,” and “general timeline of steps” illustrate when sterilization takes place in both the setup of the bioreactor and when performing our cell study. Sterilization consists of autoclaving the bioreactor mounts and tubing, UV exposure of the entire bioreactor with all components within a biosafety cabinet , and ethanol washing or adapters, fittings, and tubing. This test consists of measuring glucose concentration and pH within the feed reservoir and the output of each cell culturing chamber. In addition to this qualitative portion, we will inspect for microbes within the cell chambers. These three measurements will be performed at different time points throughout a 5 day timespan. Furthermore, for this test, we will use a typical media formulation of DMEM/10% FBS / 1% P/S.
Sterility Test Protocol:
Autoclave materials
Tubing
Microfluidic Chips
PDMS
Media Bottles
Use 70% ethanol to wash adapters and fittings by submerging the parts and using a syringe to push EtOH through the parts. Perform this step for at least 30 minutes.
Allow parts to air dry within a biosafety cabinet under UV light exposure. Perform this step for at least 1 hour, but preferably overnight.
Create the media = DMEM + 10% FBS + 1% P/S
Sterilize filter media and store until use.
Once the autoclaved parts are complete and all parts are dry, assemble a bioreactor within a BSC.
Place media within each cell chamber and inside a petri dish
Image cell chambers containing media for initial microbial inspection.
Place cell chambers back into BSC by discarding the petri-dishes.
Prime the system and flush with the media within a BSC.
Retrieve initial samples of media within the reservoir and at the output.
Place the bioreactor within an incubator.
Run the pump at 30 uL/min.
Retrieve media at the following times:
From each of these measurements, we can determine the glucose and pH difference between the media reservoir and the output of each chip. This will help determine potential contamination and microbial growth. Furthermore, imaging each chip until five days surpass allows us to visually determine if contamination does occur.
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