Table 1: Basic assays for MSC-based products.
Specifications |
Assays |
Criteria |
|
Positive |
Negative |
||
Donor screening |
Testing for virus and other communicable disease |
N/A |
HTLV-1, HTLV-2 and
CMV, HIV-1, HIV-2, HBV, HCV, Treponema pallidum, human transmissible spongiform encephalopathy
(including CJD), and other communicable disease associated with
xenotransplantation |
Viability test |
Viability Assay |
> 70% |
N/A |
Purity test |
Residual contaminants tests |
N/A |
Residual
peptides, proteins, and reagents |
Pyrogenicity/endotoxin tests |
Endotoxin:< 5 EU/kg body
weight/dose (for intrathecally administered drugs
is < 0.2 EU/kg body weight/dose) |
||
Safety test |
Bacterial and fungal test |
N/A |
Bacteria and fungi |
Mycoplasma test |
N/A |
Mycoplasma |
|
Adventitious viral agent test |
N/A |
Virus |
|
Tumorigenicity assays (karyotype analysis, teratoma
formation assay, soft agar assay, CGH, FISH, and PCR) |
N/A |
Chromosomal
abnormalities, teratoma formation, colony formation
|
|
Identity test |
Immunophenotypic profiles |
High expression (95%) of CD105, CD73, and CD90 |
Lack of expression (< 2%) of CD45, CD34, CD11b,
CD14, CD38, CD19, CD31, and HLA-DR |
Potency test |
Multilineage differentiation ability assays |
Osteogenic lineages |
N/A |
Adipogenic lineages |
N/A |
||
Chondrogenic lineages |
N/A |
||
Secretion Profiles |
FGF-2, HGF, EGF, LIF and others |
N/A |
|
CFU-F assay |
Colony formation |
N/A |
|
Immunosuppressive
assay |
Surface molecules and co-stimulatory molecules |
N/A |
Abbreviations: CFU-F: Colony-forming unit – fibroblast; CGH:
Comparative Genomic Hybridization; CJD: Creutzfeldt-Jakob Disease; CMV:
Cytomegalovirus; EGF: Epidermal growth factor; FGF-2, Basic fibroblast growth
factor-2; FISH: Fluorescence in situ hybridization; HBV, HCV: Hepatitis B and C
viruses; HGF, Hepatocyte growth factor; HIV-1, HIV-2: Human immunodeficiency
virus types 1 and 2; HLA-DR, Human leukocyte antigen - antigen D related;
HTLV-1, HTLV-2: Human T-lymphotropic virus types 1
and 2; LIF, Leukemia inhibitory factor; N/A: Not Applicable; PCR: Polymerase
chain reaction