Sample Volumes Required for Biosafety Testing (Cell Bank Sterility)
The following tables state our protocol references and the minimum amount of sample needed to carry out the assay in question.
We can also amplify test cell lines in order to generate a sufficient quantity for various tests required. In such a case, MDS requires access to the client's experimental details (SOPs) and media for cell expansion.
Isoenzyme, sterility, mycoplasma testing
Test |
Supernatant |
Cells |
Duration (weeks) |
Isoenzyme analysis |
|
2X1 mL at 107/mL |
1 |
B&F direct inoculation |
15 mL |
3 x 1 mL at 5x 106/mL |
3 |
Mycoplasma (PTC) |
25 mL |
25 mL at 103/mL+2 X T 75 Flasks |
5 |
In vitro testing
Test |
Supernatant |
Cells |
Duration (weeks) |
Using a sensitive MRC5 cell line |
8 mL |
5 mL at 107 /mL + 5 mL supernatant |
4 |
Using a sensitive Vero cell line |
8 mL |
5 mL at 107 /mL + 5 mL supernatant |
4 |
Using a sensitive BHK-21 cell line |
8 mL |
5 mL at 107 /mL + 5 mL supernatant |
4 |
Using a sensitive PPK cell line |
8 mL |
5 mL at 107 /mL + 5 mL supernatant |
4 |
Using a sensitive BT cell line |
8 mL |
5 mL at 107 /mL + 5 mL supernatant |
4 |
Electron microscopy
Test |
Supernatant |
Cells/mL |
Duration (weeks) |
TEM |
2X15 ML |
2X1 mL at 107/mL |
2 |
TEM negative stain |
2X15 ML |
2X1 mL at 107/mL |
2 |
Retrovirus testing
Test |
Supernatant |
Cells/mL |
Duration (weeks) |
F-PERT assay |
15 mL |
2X1 mL at 107/mL |
2 |
Extended assay for murine ecotropic retroviruses by co-cultivation on NIH 3T3 over 3 passages and Fg10 S+L- assay |
15 mL |
5 x 106/mL |
3 |
Extended assay for murine xenotropic/amphotropic/MCF retroviruses by co-cultivation on Mus dunni over 3 passages and PG4 S+L- assay |
15 mL |
5 mL at 2 x 107/mL + 5 mL supernatant |
3 |