Recurrent genomic instability of chromosome 1q in neural derivatives of human embryonic stem cells
Christine Varela, Jérôme Alexandre Denis, Jérôme Polentes, Maxime Feyeux, Sophie Aubert, Benoite Champon, Geneviève Piétu, Marc Peschanski, Nathalie Lefort
Christine Varela, Jérôme Alexandre Denis, Jérôme Polentes, Maxime Feyeux, Sophie Aubert, Benoite Champon, Geneviève Piétu, Marc Peschanski, Nathalie Lefort
View: Text | PDF
Research Article

Recurrent genomic instability of chromosome 1q in neural derivatives of human embryonic stem cells

Abstract

Human pluripotent stem cells offer a limitless source of cells for regenerative medicine. Neural derivatives of human embryonic stem cells (hESCs) are currently being used for cell therapy in 3 clinical trials. However, hESCs are prone to genomic instability, which could limit their clinical utility. Here, we report that neural differentiation of hESCs systematically produced a neural stem cell population that could be propagated for more than 50 passages without entering senescence; this was true for all 6 hESC lines tested. The apparent spontaneous loss of evolution toward normal senescence of somatic cells was associated with a jumping translocation of chromosome 1q. This chromosomal defect has previously been associated with hematologic malignancies and pediatric brain tumors with poor clinical outcome. Neural stem cells carrying the 1q defect implanted into the brains of rats failed to integrate and expand, whereas normal cells engrafted. Our results call for additional quality controls to be implemented to ensure genomic integrity not only of undifferentiated pluripotent stem cells, but also of hESC derivatives that form cell therapy end products, particularly neural lines.

Authors

Christine Varela, Jérôme Alexandre Denis, Jérôme Polentes, Maxime Feyeux, Sophie Aubert, Benoite Champon, Geneviève Piétu, Marc Peschanski, Nathalie Lefort

×

Figure 3

Differentiation potential in vitro and in vivo.

Options: View larger image (or click on image) Download as PowerPoint
Differentiation potential in vitro and in vivo. (A–F) In vitro neuronal ...
(A–F) In vitro neuronal differentiation potential. 3 weeks after growth factor withdrawal, normal VUB01 passage 18 NSCs (A), normal VUB05-HD passage 15 NSCs (B), and mutant VUB05-HD (batch b) passage 61 NSCs (C) differentiated in vitro into TUBB3-positive neurons. (D) Number of neurons generated with normal VUB01 and VUB05 and mutant VUB05-HD (batch a and b) NSC lines, determined using the neuronal nuclear marker HuCD. Samples of normal and mutant cell lines were differentiated in at least 3 independent experiments. The proportion of HuCD-positive cells (± SEM) was determined in at least 1,000 cells per sample in randomly picked fields. NSC lines bearing 1q duplication failed to differentiate into neurons, except mutant VUB05-HD (batch b). (E and F) 2 weeks after growth factor withdrawal, normal VUB01 passage 18 NSCs (E) differentiated in vitro into neurons, whereas mutant VUB01 passage 71 NSCs (F) failed to give rise to neurons and died. (G and H) In vivo differentiation potential. 7 weeks after grafting, human nestin- and MAP2-positive cells were observed with rat brain transplants of normal VUB01 passage 21 NSCs (G), but not with mutant VUB01 passage 74 NSCs (H). Scale bars: 50 μm.