After you’ve used our printing parameters guide for your biomaterial of choice or you’ve completed the biomaterial optimization yourself, you are ready to start bioprinting with cells! Before you start, there are a few steps you should follow. Read on for our guide to bioprinting experiment preparation.
Experiment Prep: Clean your workspace
While working with live cells during the bioprinting experiment you should take extra precautions to avoid contamination. Start with preparing your workspace. We recommend putting Allevi bioprinters into a cell culture hood (laminar flow cabinet), which will provide a sterile environment throughout the bioprinting process. Since there is significant interaction with the bioprinter, sample, biomaterials, and cells we designed our printers to be easily accessible and open.
- Clean the cell hood according to your equipment manufacturer’s instructions. Wiping surfaces with ethanol and/or UV irradiation for 30 minutes is sufficient.
- Clean the Allevi bioprinter. Gently spray visible surfaces using 70% ethanol and wipe them with a clean paper towel. Make sure to reach all corners, including the power cable, USB cable, and compressor hose. Do not immerse any parts in the ethanol or any other liquid to avoid damaging electronic components.
- Put the Allevi bioprinter inside the clean cell hood.
- Optional: UV irradiation on the printer for an additional 30 minutes.
- Make sure that after installation the airflow in the cell hood is at the operational level.
Experiment Prep: Prepare your bioink
For printing live cells encapsulated in a biomaterial, there are several points to consider:
- Is your biomaterial sterile? In our shop, we offer some sterile biomaterials, but sometimes when you prepare the hydrogel yourself, you might need to sterilize it before. For most hydrogels, heat treatment is not suitable, because it can change biological and mechanical properties, thus we recommend filtration.
- Select your filter:
- Simply aspirate your hydrogel to a syringe with Luer-lock fitting. Next, attach the appropriate filter and push the contents of the syringe through it. Collect the flow-through in a sterile tube.
- Notes: Make sure your biomaterial is at the right temperature. Some will be liquid in 4 °C (e.g. Pluronic), others will start cross-linking at 37 °C (e.g. collagen).
- Some protein-rich biomaterials might not be suitable for filtering, due to a loss of protein during the process. Some other methods such as ethylene oxide treatment or irradiation can be more suitable.
- Sterilize your consumables
We offer sterile plastic 5 ml syringes and 10 ml syringes. However, if you purchased a non-sterile version and you would like to use them with cell-laden hydrogels, autoclaving will be necessary. The same applies to our plastic needles and metal nozzles.
- To sterilize needles or syringes use a standard autoclaving program at 121°C, 15 PSI pressure for a minimum of 30 minutes. In the case of syringes, remember to remove the plunger before autoclaving. You can use a sterilization pouch for easy handling of the sterilized products.
- For non-Allevi products, check the manufacturer’s information.
- Combine cells with your biomaterial
- Use our guide for mixing your cells with your biomaterial.
The keys to a successful bioprinting experiment are having a sterile environment, sterile biomaterial, and a good experimental plan.
If you have any questions regarding the bioprinting experiment preparation protocol above, please contact us at [email protected]