High density (HD) cell banking is seed train optimization involving larger volume frozen seed banks, eliminating seed steps from vial and enabling banking of larger volumes up to 1 liter.

HIGH DENSITY CELL BANKING

For the production of biopharmaceuticals, a seed train is required to generate an adequate number of cells for inoculation of the production bioreactor. Conventional seed train processes begin with thawing of a cryopreserved cell-bank vial, followed by successive expansions into larger culture vessels.  When culture volume and cell density meet pre-determined criteria, the culture is transferred to a production bioreactor in which cells continue to grow and express product.

Conventional seed train starting from a vial is time- and cost-intensive.

  • Shake flasks or spinners used in the initial stages require manual manipulations inside a laminar flow hood, making them vulnerable to contamination and operator error.
  • Because a conventional cryopreserved cell-bank vial contains low cell numbers, the seed-train process is time consuming.
  • If the production bioreactors are very large, additional large-volume culture vessels are required to support scale up to a production bioreactor

High density (HD) cell banking is seed train optimization involving larger volume frozen seed banks, eliminating seed steps from vial and enabling banking of larger volumes up to 1 liter.

  • Consistent seed inoculations across different process runs
  • Reduced seed train process complexity by decreasing the number of lab-scale expansion stages
  • Eliminates the need for labor-intensive operations such as cleaning, assembly, and sterilization 

 

 

 

by Benjamin Wright, Mike Bruninghaus, Mike Vrabel, Jason Walther, Neha Shah, Seul-A Bae, Timothy Johnson,Jin Yin, Weichang Zhou and Konstantin Konstantinov
BioProcess International
March 2015

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A Novel Seed-Train Process: Using HD Cell Banking, a Disposable Bioreactor, and Perfusion Technologies

THE XCell™ ATF SYSTEM IN HIGH DENSITY CELL BANKING

 
XCell™ ATF Cell Retention System
  • Continuous, provides clarified harvest ready for capture chromatography
  • Efficient, delivers 100% cell retention, increases productivity
  • Convenient single-use format reduces implementation time by up to 80%
  • Intensified, generates high cell concentration and viability
  • Flexible, connects to any bioreactor and can be used in a variety of applications
  • Gentle, reduces cell stress, increases cell viability and productivity
  • Easy to use, reduces setup time and user training
  • Scalable, adapts from development-scale to production-scale
  • Self-cleaning, reduces biofilm build up on the hollow fiber filter

Learn more about the XCell™ ATF System

Using the XCell™ ATF System produced consistent seed quality, reducing growth time by up to 10 days

Create a Cryo-seed bank

This XCell™ ATF perfusion was conducted to generate 100E6 cells/mL VCD and stored in 100mL and 1L HDCSI cryo-bags (5% DMSO and 0.11% CMC were used in conditioned media as cryo-protectants)

n-1 Perfusion using Cryo-bag (100mL) seed

Cells from cryo-bags were used to inoculate n-1 reactor at 1E6 cells/mL VCD, produces consistent and reproducible cell growth

n-1 Perfusion comparing Cryo-bag 100mL and 1L

Comparison of 100mL and 1L cryo bag. The recovery and growth curve profiles are similar between 100mL cryo-bag thaw compared to 1L cryo-bag thaw

High density cell banking, using the XCell™ ATF System attached to high density cryo-bags, helps eliminate the seed step from vial, and enables banking of larger volumes up to 1 liter.

see poster

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