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)
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 TM to 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 TM manually 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 TM manually 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.
