Table 2.
MSC culture process validation
| (a) Cultures rates | |||||
|---|---|---|---|---|---|
| 1st Exp. | 2nd Exp. | 3rd Exp. | |||
| BM sample (mL) | 28 | 52 | 55 | ||
| MNC on day 0 | 180 × 106 | 195 × 106 | 260 × 106 | ||
| MSC on day 28 | 225 × 106 | 165 × 106 | 356 × 106 | ||
| (b) Phenotypes | |||||
|---|---|---|---|---|---|
| Marker | Eligibility criteria (%) | 1st Exp. (%) | 2nd Exp. (%) | 3rd Exp. (%) | Conformity |
| CD73 | >70 | 96.9 | 99.9 | 99.4 | Ok |
| CD105 | >70 | 85.6 | 90.3 | 77.8 | Ok |
| CD90 | >70 | 100 | 98.2 | 100 | Ok |
| CD34 | <10 | 1.8 | 8.8 | 0.92 | Ok |
| CD45 | <10 | 1.1 | 1.10 | 0.55 | Ok |
| HLA-DR | <10 | 0.84 | 0.63 | 0.52 | Ok |
| CD80 | <10 | 1.0 | 0.7 | 0.45 | Ok |
| CD31 | <10 | 1.0 | 0.9 | 0.62 | Ok |
| (c) Post-thaw | |||||
|---|---|---|---|---|---|
| Test | Eligibility criteria | 1st Exp. | 2ndExp. | 3rd Exp. | Conformity |
| Initial cell number | Na | 100 × 106 | 70 × 106 | 55 × 106 | Na |
| Initial viability | Na | 95.8 % | 82.5 % | 86.0 % | Na |
| Post-thaw cell count | Na | 100 × 106 | 56 × 106 | 43 × 106 | Na |
| Post-thaw cell viability | >50 % | 89.5 % | 78.0 % | 78.0 % | Ok |
| Post-thaw cell culture expansion | Na | Ok | Ok | Ok | Na |
| Sterility | Sterile | Ok | Ok | Ok | Ok |
| Mycoplasma | Absence of mycoplasma | Ok | Ok | Ok | Na |
| Endotoxin | Endotoxin level <2.5 U.I./ml | Ok | Ok | Ok | Na |
| Phenotype | |||||
| CD73 | >70 % | 99.5 % | 96.1 % | 87.9 % | Ok |
| CD105 | >70 % | 93.6 % | 70.3 % | 70.0 % | Ok |
| CD90 | >70 % | 100 % | 99.6 % | 99.8 % | Ok |
| CD34 | <10 % | 3.1 % | 2.7 % | 2.6 % | Ok |
| CD45 | <10 % | 1.0 % | 1.9 % | 1.0 % | Ok |
| HLA-DR | <10 % | 1.0 % | 1.2 % | 1.0 % | Ok |
| CD80 | <10 % | 0.4 % | 0.8 % | 0.4 % | Ok |
| CD31 | <10 % | 0.6 % | 1.2 % | 0.6 % | Ok |
Results of three large-scale MSC cultures for initial process validation: culture rates, phenotypes at final harvest and post-thaw parameters are reported