Table 1.
Quality control and preclinical research of cell line HAMSC10.
| Measurement indicators | Testing items | Quality standards | Results |
|---|---|---|---|
| (1) Cell identity | |||
| Cell morphology (QCP3−QCP6) | Cell shape | Fusiform | Up to standard |
| Genetic characteristics (QCP5) | Chromosome karyotype | 46, XY/46, XX | 46, XY |
| STR map | Single origin | Up to standard | |
| Cell surface antigens (QCP3−QCP6) | CD73, CD90, CD105, CD44 | Positive cells ≥ 95% | Up to standard |
| CD45, CD34, CD11b, CD19, HLA-DR | Positive cells ≤ 2% | Up to standard | |
| (2) General biosafety | |||
| Common microorganism contamination (QCP2−QCP6) | Fungus, bacteria, mycoplasma, Treponema pallidum | Negative | Up to standard |
| Endotoxin | <0.5 EU/ml | Up to standard | |
| (3) Endogenous and exogenous pathogenic factors | |||
| Detection of virus infection by molecular analysis (QCP3 and QCP5) | HIV-1, HBV, HCV, EBV, HCMV, HHV6/7, HPV, retrovirus | Negative | Up to standard |
| Detection of virus infection by cell culture (QCP5) | Stem cell culture test | Normal morphology | Up to standard |
| Hemadsorption/hemagglutination test | Negative | Up to standard | |
| Detection of virus infection by animal inoculation (QCP5) | Guinea pig inoculation testa | Negative | Up to standard |
| (4) Cell viability and growth characteristics | |||
| Cell viability (QCP2−QCP6) | Living cell counting | ≥95% | Up to standard |
| Growth curve analysis (QCP6) | Cell population doubling time | Report result | 32.8 h |
| Cell cycle analysis (QCP5) | Cells in S-stage | Report result | 12.8% |
| (5) Tumorigenicity | |||
| In vitro tumorigenicity (QCP5) | Colon formation test, animal hypodermic inoculation test | Negative | Up to standard |
| In vivo tumorigenicity (QCP5) | Subcutaneous test in nude mice | Negative | Up to standard |
| (6) Biological potency | |||
| Pluripotency (QCP5) | Adipogenic differentiation, osteogenic differentiation, chondrogenic differentiation | Positive | Up to standard |
| Cytokine secretory function (QCP5) | HGF | Report result | >1000 pg/ml |
| BDNF | Report result | >100 pg/ml | |
| Nerve growth factor | Report result | >10 pg/ml | |
| Immunological effect (QCP5, AMSCs:PBMCs = 1:10) | Lymphocyte proliferation inhibition test | Report result | Lymphocyte proliferation inhibited |
| Special lymphocyte subpopulation proliferation test (Th1/Th17/Treg) | Report result | Treg increased, Th1 and Th17 decreased | |
a
Test methods were according to the Pharmacopoeia of People's Republic of China (2015 version, part III) except for that specially described in the Materials and methods section.
QCP, quality control point; STR, short tandem repeat; HIV-1, human immunodeficiency virus-1; HBV, hepatitis B virus; HCV, hepatitis C virus; EBV, Epstein–Barr virus; HCMV, human cytomegalovirus; HHV6/7, human herpes virus 6/7; HPV, human papillomavirus; PBMCs, peripheral blood mononuclear cells.