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Note:
·
Up to 5 billion T-cells have been loaded in the CFC Chamber using high G and a very low flow rate
but 2 – 3 billion T-cells is more common.
·
Cells <4 μm, e.g. platelets, cannot be concentrated in the CFC Chamber and hence do not need to
be considered when calculating the volume of input material that can be loaded.
·
Elutriation requires a lower G force and higher flow rate than loading, reducing the concentration and
hence number of cells in the fluidized bed.
·
When loading cells prior to an elutriation step, be sure to allow enough room in the CFC Chamber to
retain the larger cells as the fluidized bed expands.
Bed disruption during loading
In certain conditions, low density immiscible media may accumulate as a separate layer of fluid in the
input bag. When this fluid enters the CFC Chamber, it is highly buoyant and can disrupt the established
fluidized bed. It is important to keep materials of this nature mixed during loading.
Cell separation during loading
While it is common practice to be conservative during cell loading to avoid losses of target cells, the
loading step can be an efficient way to remove debris and lighter cell materials. For example, this
strategy can be used during leukopak processing where debris and platelets are washed out as the
PBMC is loaded.
CTS
™
Rotea
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Counterflow Centrifugation System Process Design User Guide
Chapter 4 Protocol Building Blocks
Cell loading
4
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