Table 1.
Immunophenotyping, apoptotic index and distribution of neurogenic markers on unpurified mixed population
| MSCs Day < 0 |
NSPs Day 30 |
NLCs Day 37 |
|
|---|---|---|---|
| Differentiation of hADSCs in healthy subjects | |||
| CD34(+) | 1.26 ± 2.01 | 0.18 ± 1.64 | 0.51 ± 2.60 |
| CD45(+) | 1.28 ± 1.17 | 0.71 ± 0.92 | 0.07 ± 1.34 |
| CD73(+) | 90.12 ± 5.52 | 24.51 ± 10.34 * | 6.77 ± 3.68 ** |
| CD90(+) | 95.59 ± 6.80 | 4.07 ± 3.76 ** | 1.94 ± 2.1 *** |
| CD105(+) | 84.33 ± 5.33 | 18.68 ± 4.51 * | 1.26 ± 1.90 *** |
| Annexin V(−)/PI(−)/7AAD(−) (viable cells) | 98.97 ± 0.71 | 96.20 ± 1.25 | 96.85 ± 0.83 |
| Annexin V(+)/PI(−)/7AAD(−) (early-apoptotic cells) | 0.76 ± 0.90 | 2.11 ± 1.01 | 2.03 ± 1.04 |
| Annexin V(+)/PI(+)/7AAD(+) (late apoptotic, necrotic cells) | 0.27 ± 0.71 | 1.69 ± 0.99 | 1.12 ± 1.10 |
| NeuN | 1.62 ± 1.54 | 58.14 ± 3.32 ** | 78.12 ± 6.28 ** |
| TUJ1 | 4.49 ± 1.62 | 60.48 ± 5.25 ** | 89.09 ± 8.67 ** |
| O4 | 1.28 ± 2.40 | 28.16 ± 4.11* | 26.33 ± 3.29 * |
| GFAP | 24.14 ± 2.58 | 4.26 ± 1.84 ** | 1.20 ± 1.24 ** |
| Nestin | 54.01 ± 8.41 | 20.15 ± 2.64 * | 0.81 ± 2.62 ** |
| Differentiation of hADSCs in ND subjects | |||
| CD34(+) | 1.82 ± 1.72 | 0.08 ± 1.21 | 0.71 ± 1.05 |
| CD45(+) | 3.69 ± 1.88 | 0.90 ± 1.14 | 0.11 ± 1.92 |
| CD73(+) | 79.49 ± 3.42 | 4.06 ± 2.66 ** | 2.90 ± 2.03 *** |
| CD90(+) | 93.51 ± 3.01 | 0.73 ± 1.40 *** | 0.98 ± 1.74 *** |
| CD105(+) | 86.28 ± 6.76 | 13.53 ± 5.18 ** | 5.04 ± 7.18 ** |
| Annexin V(−)/PI(−)/7AAD(−) (viable cells) | 97.12 ± 1.13 | 93.10 ± 2.44 | 98.33 ± 2.91 |
| Annexin V(+)/PI(−)/7AAD(−) (early apoptotic cells) | 1.96 ± 0.76 | 4.71 ± 1.82 | 0.02 ± 2.16 |
| Annexin V(+)/PI(+)/7AAD(+) (late apoptotic, necrotic cells) | 0.92 ± 1.33 | 2.19 ± 1.37 | 1.65 ± 0.24 |
| NeuN | 0.37 ± 1.09 | 45.20 ± 2.07 ** | 50.31 ± 2.53 **, § |
| TUJ1 | 1.94 ± 1.13 | 40.11 ± 4.17 **, § | 52.65 ± 6.24 **, §§ |
| O4 | 4.13 ± 5.10 | 38.14 ± 5.23** | 18.04 ± 1.88 *, § |
| GFAP | 34.05 ± 3.67 § | 4.19 ± 2.12 ** | 2.43 ± 4.16 ** |
| Nestin | 73.56 ± 7.82 § | 16.20 ± 3.75 ** | 1.05 ± 2.35 ** |
Cells were labeled with fluorescence-coupled antibodies against CD34, CD45, CD73, CD90, CD105, Annexin V, 7AAD, propidium iodide solution (PI) and neural markers and analyzed using a MACSQuant flow analyzer as indicated in Methods. Results presented as mean ± SEM and presented as the percentage of cell surface marker per cell type (% of total cells) of all subjects each performed in duplicate in each group. Cells collected at day 0 before the induction of the differentiation (MSCs) and days 30 and 37 of differentiation (NSPs and NLCs, respectively)
hADSC human adipose-derived mesenchymal stem cell, MSC mesenchymal stem cell, ND neurodegenerative disease, NLC neural-like cell, NSP neurosphere-like structure
*P < 0.05, **P < 0.01 and ***P < 0.005: NSPs/NLCs vs MSCs
§P < 0.05, §§P < 0.01: ND vs healthy