Cultivation Parameters; Sampling And Inoculation; Preparing Sampling Or Inoculation - Sartorius Stedim Biotech BIOSTAT RM 20 basic Bedienungsanleitung

4.5 Cultivation Parameters

4.6 Sampling and Inoculation

4.6.1 Preparing Sampling
or Inoculation
Each cell line requires optimization of the cultivation conditions.
The following table contains recommendations for the aeration rate and the
rocker angle to start the cultivation process (room air with an oxygen content
of 20 %).
Liquid volume Rocker speed
(L)/CultiBag RM [r/min]
1/2 15–20
5/10 15–20
10/20 15–20
25/50 15–20
Air intake (aeration rate) and CO
An oversized air intake only has a minimal effect on the oxygen intake. The aeration
rate should be kept to a minimum to reduce the evaporation of the medium. The
recommended aeration rate is 0.1–0.2 l/min. We recommend a CO
5 % because this is used for most mammalian cells. However, this must be adjusted to
each cell line and medium.
Rocker speed
The rocker speed must be set to the minimum speed that ensures thorough mixing
and oxygen intake without leading to excessive foam buildup and the associated
shearing forces. At the start, the rocker speed is normally 15–20 rocker movements/
min. The rocker speed required for the oxygen intake is designed for the respective
cell population and its metabolism. Normally, 15–25 rocker movements/min. are
enough to ensure the oxygen requirement of cell thicknesses up to 5 + 10
Rocker angle
A smaller rocker angle can minimize foaming of the medium. The rocker angle can be
reduced 2–3° for high foaming media. The rocker angle can be increased up to 8° for
cells with a high oxygen requirement, e.g. insect cells. The maximum recommended
rocker angle is 10°.
This procedure requires a standard laboratory syringe with a Luer septum without a
needle. This procedure must not be carried out on a sterile workbench.
1. Stop the rocker movement. The rocker has an easy-to-use Sampling function for
this, which is explained in more detail in section 4.4.2.
2. Do not interrupt the surface aeration.
3. Remove the protective cover from the Luer septum of the sampling tube.
Aeration rate
[l/min]
0,1
0,1–0,2
0,2
0,3–0,5
concentration
2
Menu Operation
Rocker angle
6–8
6–8
6–8
6–8
concentration of
2
6 cells/ml.
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