Figure 4. Safety profile of reversibly immortalised human mesoangioblasts.

1. In vitro serum and growth factor dependence of hTERT + Bmi1 polyclonal populations (H#1 and H#2, left graphs, n = 2 for each groups) and hTERT + Bmi1 clones (H#3A, H#3B and H#3C, right graphs, n = 4). HeLa cells: positive control (n = 4). BrdU incorporation rate was calculated as the % of BrdU‐positive cells on total number of nuclei in 1 h time. Data plotted as means ± SEM. *P = 0.0314, ***P = 0.0002, ****P < 0.0001, one‐way analysis of variance (ANOVA) with Tukey's post hoc post‐test.
2. Cell contact inhibition assays of hTERT + Bmi1 polyclonal populations (H#1 and H#2, left graphs, n = 2 for each groups) and hTERT + Bmi1 clones (H#3A, H#3B and H#3C, right graphs, n = 4). HeLa cells: positive control (n = 4). BrdU incorporation rate was calculated as the % of BrdU‐positive cells on total number of nuclei. Data plotted as means ± SEM. *P = 0.0190 (HeLa, 0 vs. 4), **P = 0.0035 (H#1, 0 vs. 4), **P = 0.0020 (H#1, 0 vs. 8), ***P = 0.0003 (H#1, 0 vs. 12), ***P = 0.0006 (H#3B, 0 vs. 8), **** P < 0.0001, ns = 0.0627 (H#3B, 0 vs. 4), ns = 0.9643 (HeLa, 0 vs. 8), ns = 0.0790 (HeLa, 0 vs. 12), one‐way analysis of variance (ANOVA) with Tukey's post hoc post‐test.
3. Karyotype analysis of immortalised mesoangioblast polyclonal populations.
4. qRT for hTERT and Bmi1 transgenes in immortalised mesoangioblasts (H#2) 2 weeks after transduction with different multiplicity of infection (MOI 0.5, 1, 2.5, 5 and 10) of IDLV NLS‐Cre. Gapdh was used as housekeeping gene. Immortalised mesoangioblasts have been used as reference (=1, black bar). Data plotted as means ± SEM (n = 2). Red box: IDLV NLS‐Cre MOI 2.5 immortalised mesoangioblasts used for further experiments with ganciclovir.
5. hTERT and Bmi1 PCRs on MOI 2.5 IDLV NLS‐Cre‐transduced immortalised mesoangioblasts treated with 10 and 20 μM ganciclovir. Positive control: untreated (−) immortalised mesoangioblasts.

Source data are available online for this figure.