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Media density (ρ)
It is important that there is enough difference in density between the cells (ρ
Note: If the media is denser than the cells, then the cells cannot settle and a bed will not form.
Even very subtle changes in media density can affect the stability of the bed and the settings for
loading and separating cells. The higher the density of the media, the higher the G force or lower the
flow rate needed to form a bed.
Cell concentration
The number of cells required for a fluidized bed to form is strongly influenced by the cell diameter (d)
and the density differential (ρ
• MSC's: >20 × 10
• PBMC's: >30 × 10
• When the cell concentration in the input material is low, it will take longer to reach the cell
concentration in the CFC Chamber and hence for the fluidized bed to form.
• While the Rotea
beds are much more sensitive to changes in conditions. Where possible, it is best to work with
larger numbers of cells
Establish a bed in recirculation
Under most situations it is preferred to establish a bed in such a way that no cells are lost to the waste
bag during the formation of the bed. This technique is particularly important when the cells are small,
the input concentration low or the media dense.
A dual-connection input bag contains the cells in media. At the define kit screen, define a bag with two
connections. The Rotea Protocol Builder will give you the available second connection options.
CTS
™
Rotea
™
Counterflow Centrifugation System Process Design User Guide
– ρ
) but as a guide:
p
m
6
cells
6
cells
™
instrument is capable of processing comparatively small numbers of cells, small
Chapter 4 Protocol Building Blocks
Establish a fluidized bed
) and the media (ρ
p
4
).
m
57
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