What you need to know about large scale cultivation (Issue 4) —Tutorial on Operating BioFactories in Series-무석내사생명과학기술주식유한회사

Cell culture is a complex and time-consuming process which requires significant efforts. In the beginning, the cells are nurtured in a cell culture plate or flask, but as they grow, they needed to be transferred to larger containers. This is where multiple BioFactories ^TM connection comes in. In the previous issue, we have discussed the advantages of closed systems, how to customize a desired one, as well as the ways to prevent contamination. In this issue, we will introduce the accessories available for BioFactory ^TM, how to make the most of BioFactory ^TM and its accessories, and how to connect individual "villas" into "townhouses" like toys so as to boost laboratory efficiency.

Part One NEST Supportive Gadgets - Bioprocessing Accessories

Liquid Transferring Hoses

Connectors

Other

Part Two Figure of BioFactory ^TM Connection

Due to the feasibility of connection between BioFactories ^TM, the culture process can be scaled up by simply using more BioFactories ^TM, which may increases the risk of contamination caused by repeated opening operations. However, there is a simple solution to this problem: connect BioFactories ^TM together into a semi-closed/closed system with accessories.

Tutorial on operating three 40-layer BioFactories ^TM in series (demonstration of one method)

Part Three Tutorial on connecting BioFactory ^TM (taking a 10-layer BioFactory ^TM as an example)

Seeding and Innoculation

1. Connect the BioFactory ^TM in series first. Install the hose from Carboy to the peristaltic pump, ensuring that the hose is properly secured and connected to the tubing of the BioFactories ^TM in series.

2. Open all the clamps on the hose and turn on the peristaltic pump. The culture medium is pumped into the connected BioFactories ^TM via the hoses. Try to prevent bubbles or foams during operation.

Note: The inflow amount can be controlled by closing the corresponding clamp since the rate may vary.

3. When getting a sufficient inflow amount, turn off the peristaltic pump and close all the clamps.

4. Take the hose from the peristaltic pump and disconnect it from the tubing. Place the BioFactory ^TM on its side to evenly distribute the liquid to each layer. Make sure the filter side is on the top while the inlet side is at the bottom to prevent liquid from entering the filter.

5. Rotate 90 degrees clockwise with the inlet side facing upwards, and let it stand for a while to distribute the liquid evenly to each layer. Slowly place the BioFactory ^TM horizontally, tidy up the tubing or replace it with a vent cap (or a filter), and then put it in the incubator for incubation.

Note: If the end of the tubing is heat-sealed, connection or disconnection process can be performed under normal environment conditions. If the end of the tubing is a joint, perform the connection or disconnection process in a Class 100 cleanroom.

Collection

1. Connect the BioFactory ^TM in series and place them laterally (the tubing side at the bottom and the filter on the top). Install the hose from the Carboy to the peristaltic pump, ensuring that the hose is properly secured and connected to the tubing of the BioFactories ^TM.

2. Open all clamps and turn on the peristaltic pump. Collect the culture medium into the Carboy via the hose.

3. Tilt the BioFactory ^TMto facilitate further collection after collecting a certain amount of fluid.

Note: It is recommended to raise the BioFactory ^TM on a rack for collection. If there is no rack, raise the BioFactory ^TMmanually to ensure complete pouring out.

Digestion

1. Connect the hose to the Carboy containing digestion fluid and start the peristaltic pump to fill in the digestion fluid.

Note: Since the inflow rate may vary, the inflow amount can be controlled by closing the corresponding clamps.

2. Turn off the peristaltic pump and take the hose from it. Place the BioFactory ^TM laterally to ensure that the digestion fluid is evenly distributed to each layer. Make sure the filter side is on the top while the inlet side is at the bottom to prevent liquid from entering the filter.

3. Rotate the BioFactory ^TM 90 degrees clockwise with the inlet side facing upwards, and let it stand to distribute the liquid evenly to each layer.

4. Slowly place the BioFactory ^TM horizontally. Gently shake each BioFactory ^TM to ensure that the digestion fluid covers the adherent cells, and then allow it to stand still for a while . After the digestion is completed, connect the tubing to the collection container, open all the clamps, and start the peristaltic pump. Tilt the BioFactory ^TM to transfer.

Note: It is recommended to raise the BioFactory ^TM on a rack to avoid residue during collection. If there is no rack, raise the BioFactory ^TMmanually to ensure complete pouring out.

5. After the collection is completed, turn off the peristaltic pump. Due to differences in production processes, different connection method and various accessories may be deployed for large-scale cell culture in BioFactory ^TM.

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