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Seed Train Optimization in Microcarrier-Based Cell Culture Post In Situ Cell Detachment through Scale-Down Hybrid

Atefeh Ebrahimian^1,2,3, Mona Schalk^1,2, Mark Dürkop⁴

¹ACIB-Austrian Centre of Industrial Biotechnology, Krenngasse 37, 8010 Graz, Austria.

Bioengineering (Basel, Switzerland)

|March 27, 2024

Summary

This study introduces a new method for cell culture using microcarriers (MCs) and in situ detachment, improving efficiency for antigen and viral vaccine production. MA 104 cells can be subcultured five times on MCs, optimizing seed training in bioreactors.

Keywords:

MA 104 cells hybrid modeling in situ cell detachment microcarrier culture process optimization seed training

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Area of Science:

Biotechnology
Cell Culture Engineering
Bioprocess Development

Background:

Microcarrier-based cell culture is standard for anchorage-dependent cells like MA 104 in antigen manufacturing.
Conventional methods involve static cell propagation before bioreactor seeding, which is inefficient.

Purpose of the Study:

To replace conventional static cell propagation with serial subculturing on microcarriers (MCs) using in situ cell detachment.
To optimize critical process parameters (CPPs) for efficient cell growth and seed training in stirred tank reactors.

Main Methods:

Serial subculturing of MA 104 cells on Cytodex 1 MCs with in situ detachment in closed systems.
Utilized augmented Design of Experiments (DoE) and hybrid modeling for CPP screening and optimization.
Investigated process parameters like inoculation density, MC concentration, and agitation speed (Njs).

Main Results:

MA 104 cells were successfully subcultured at least five times on Cytodex 1 MCs without cell-MC separation.
Optimized conditions yielded a cell density of 2.6 ± 0.5 × 10^6 cells/mL after five days.
Identified optimal inoculation density (>16 cells/bead), Cytodex 1 concentration (3.5-4.5 g/L), and agitation strategy.

Conclusions:

Developed a reliable and efficient bioprocess for seed training in stirred tank reactors.
The in situ detachment method enhances microcarrier-based cell culture for viral vaccine production.
Optimized CPPs ensure high cell density and reproducible results.