Cells and microbeads were inoculated at high concentrations and then diluted, stirred and sampled as required.

Operation method

microcarrier

Principle

Cells and microbeads were inoculated at high concentrations and then diluted, stirred and sampled as required.

Materials and Instruments

Starter Cultures
Growth medium Microcarriers Stirred culture flasks Magnetic stirrer

Move

1. Suspend the microbeads at 2~3 g/L according to 1/3 of the desired final volume of culture solution.

2. Digest and count the cells with trypsin and inoculate the cells into the microbead suspension at 3~5 times the normal inoculum concentration.

3. Agitate the culture at 10~25 rpm for 8 h.

4. Add culture solution to reach the final volume of the microbead concentration of 0.7~1 g/L.

5. Increase the agitation speed to 60 rpm. rpm.

6. If pH drops, turn off the stirrer and incubate for 5 min to allow the microbeads to settle down, then replace 1/2~2/3 of the culture solution.

7. Collect the cells:

(a) Aspirate the culture solution.

(b) Wash the cells by sedimentation.

(c) Treat the microbeads with trypsin and EDTA.

(d) Allow the microbeads to settle.

(e) Dislodge the cells from the microbeads by rotation.

(f) Wash the cells by mixing and centrifugation.

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