An optimized HEK293T cell expansion protocol using a hollow-fiber bioreactor system

Area of Science:

• Biotechnology
• Cell Biology
• Virology

Background:

• Viral vectors are crucial for genetic material delivery, with HEK293 cells frequently used for viral particle production.
• HEK293T cells, engineered with SV40 large T antigen, are vital for high-titer viral production.
• Optimizing cell culture conditions is essential for efficient HEK293T cell expansion and viral vector manufacturing.

Purpose of the Study:

• To provide a protocol for automated HEK293T cell expansion using the Quantum® hollow-fiber bioreactor (HFB) system.
• To guide users on critical parameters for successful cell expansion within the HFB environment.
• To ensure reliable control of environmental factors including glucose, lactate, and pH.

Main Methods:

• Utilized the Quantum® hollow-fiber bioreactor (HFB) system for automated cell expansion.
• Focused on optimizing and controlling key environmental parameters: glucose, lactate, and pH.
• Developed a protocol specific to the HFB milieu, distinct from traditional tissue culture methods.

Main Results:

• Demonstrated reliable control over glucose, lactate, and pH levels during HEK293T cell expansion in the HFB system.
• Provided data reflecting the consistency of controlled environmental factors.
• Outlined procedural time points for successful cell expansion.

Conclusions:

• The Quantum® HFB system offers a reliable method for automated HEK293T cell expansion.
• Optimized control of environmental parameters ensures consistent cell culture conditions.
• This protocol facilitates efficient viral vector production by standardizing HEK293T cell expansion.