In Vitro Adventitious Agent Tests on Cells
| WHO TRS 978 Annex 3 | FDA Guidance 2010 | Ph. Eur. 5.2.3 (3 tests) | Ph. Eur. 2.6.16 | |
|---|---|---|---|---|
| Test article | 107 live cells or cell lysate, each with spent culture fluid | Cell lysate equivalent to 107 cells (for cell bank) 500 doses or 50 mL whichever is greater (for viral seeds and viral harvests) | Hemadsorbing virus test on cells • Extraneous Agent test on cell culture fluids • Co-cultivation of intact and/or disrupted cells | 500 human doses of vaccine or 50 mL of viral seeds or viral harvests whichever is greater |
| Indicator cells | At least 2 cell types • the same species and tissue type • a human diploid cell line (DCL) In some countries, cultures of another (third) cell line from a different species are required. | 3 cell types • the same species and tissue type • human diploid cells • monolayer cultures of monkey kidney cells. | At least 2 cell types As per Ph. Eur. 2.6.16 for hemadsorbing virus test and for Extraneous agents test on cell culture fluids + Co-Cultivation with human and simian cells | At least 2 cell types • continuous simian kidney • human cell cultures • If growth in an avian or mammalian cell system other than simian or human, cells of that species |
| Incubation time | For at least 2 weeks • If human or simian CMV, HDC observed for at least 4 weeks (or CMV NAAT) In some countries, a passage for an additional 2 weeks | For at least 2 weeks + subculture (an additional 2 weeks). If human or simian CMV is consideration, the first set of human cell cultures observed for at least 4 weeks unless a shorter incubation period is shown to be equally sensitive. | At least two weeks as per Ph. Eur. 2.6.16 | The cells are incubated at 36 ± 1 °C and observed for a period of 14 days. |