Table 3.
Common quality control tests performed to release NSC-based products.
| Test | Method | Comments |
|---|---|---|
| Sterility | Turbidity testing after growth in TSB and TG media (Ph. Eur.2.6.1) [134] or media for automated detection systems (Ph. Eur. 2.6.27) [135] | NA |
| Mycoplasma | PCR, culture (Ph. Eur. 2.6.7) [136] | NA |
| Endotoxins | LAL assay (Ph. Eur. 2.6.14) [137] | NA |
| Adventitious viruses | PCR, cytopathic effect, others (Ph. Eur. 5.2.3 and Ph. Eur. 5.1.7) [138,139] | The viruses to detect should be evaluated case by case by risk analysis (Ph. Eur. 5.1.7) [139] |
| Cell number and Population doublings | Haematocytometer, automated cell counters, flow cytometer (Ph. Eur. 2.7.29) [140] | NA |
| Viability | Trypan blue, Calcein/Ethidium, 7AAD, PI (Ph. Eur. 2.7.29) [140] | This should be higher than 70% when transplanted [141] |
| Identity/Purity | Flow cytometry (Ph. Eur. 2.7.24) [142], PCR (Ph. Eur. 2.6.21) [143], immunofluorescence (Ph. Eur. 2.7.1) [144], others (Ph. Eur. 5.2.3) [138]. | Markers of the possible contaminant cells (e.g., PSC) should be included [145] |
| Potency | Tri-lineage differentiation potential, expression of neurotrophic factors, etc. (Ph. Eur. 2.6.34) [146] | NA |
| Tumorigenicity | Karyotype, CGH array, trophic factor dependence, colony-forming assays in softagar, in vivo tumor formation assay in athymic mice (Ph. Eur. 5.2.3) [138] | NA |
| DNA Fingerprint | STR, VNTR (Ph. Eur. 5.2.3) [138] | NA |
Selected European Pharamacopea (Ph. Eur.) chapters or guidelines that can be used as a reference have been included. TSB: Trypticase soy broth; TG: Thioglycollate; LAL: Limulus amebocyte lysate; PCR: Polymerase chain reaction; 7AAD: 7-Aminoactinomycin D; PI: Propidium iodide; CGH: Comparative Genomic Hybridization; STR: short tandem repeats; VNTR: variable number of tandem repeats; NA: not applicable.