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