Home › Cell Biology › Cultivation of Mammalian Cells Using a Single-use Pneumatic Bioreactor System
Cell Biology JoVE (Open Access) Citable · DOI

Cultivation of Mammalian Cells Using a Single-use Pneumatic Bioreactor System

DOI: 10.3791/52008-v
What you'll learn
  • ✓ Assemble and calibrate a single-use pneumatic bioreactor system
  • ✓ Inoculate mammalian cells and monitor culture parameters in real-time
  • ✓ Collect samples and characterize cell growth and viral production
Protocol

Using a pneumatic bioreactor, we demonstrate the assembly, operation, and performance of this single-use bioreactor system for the growth of mammalian cells.

Difficulty
intermediate
Total time
~7-10 days (culture duration); ~2 hrs setup and initiation per batch
Model organism
Mammalian cells (CHO, HEK293, or similar)
Biosafety
BSL-1

Steps

1
Log in and calibrate bioreactor system

Access the bioreactor control software and perform initial system calibration to ensure sensor accuracy and proper operation.

▶ 01:35
2
Install autoclave, sensors, and reagent vessels

Assemble and sterilize the bioreactor vessel, install dissolved oxygen and pH sensors, and attach reagent containers for media and supplements.

▶ 03:41
3
Add medium, microcarriers, and perform DO calibration

Introduce culture medium and microcarriers into the bioreactor, allow equilibration, and complete one-point dissolved oxygen calibration.

▶ 06:43
4
Initiate bioreactor run and inoculate cells

Start the bioreactor operation, introduce mammalian cell inoculum at target density, and begin collecting baseline samples.

▶ 08:28
5
Characterize cell culture and viral growth results

Analyze culture samples for cell viability, concentration, and viral titer to evaluate bioreactor performance and cell behavior.

▶ 10:11
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