Search

Background & Aim Genetic integrity of human pluripotent stem cells (hPSCs) is essential for the accuracy of the disease models and for the security of cell therapies that are use those cells. Because genetic abnormalities can accumulate during the generation of hPSC, their long term maintenance or during genome editing, it is mandatory to regularly check the genome of hPSCs. However, the current methods to assess genomic integrity of hPSC are not fully suitable for such regular screening. These approaches can be limited in term of resolution, flexibility, cost and time to process. Our aim is improving genetic screening of hPSC. Methods, Results & Conclusion Based on a large meta-analysis of all hPSC genetic abnormalities reported in more than 100 publications, we were able to exhaustively identify recurrent genetic abnormalities accumulating in hPSCs. We then developed a test (iCS-digital test) based on the droplet digital PCR technology that can detect more than 90% of these hPSC recurrent genetic abnormalities in DNA extracted from culture supernatant samples. This test was successfully used to detect copy number variations that arose during cell culture or gene editing. Hence, the iCS-digital test can be used to routinely screen genomic integrity in hPSCs, contribute to secure genome editing workflow, and reduce time and financial losses induced by the use of an abnormal hPSC lines.

Objet La medecine regenerative a pour ambition de regenerer des tissus endommages par la maladie ou la vieillesse. L’utilisation de cellules souches pluripotentes humaines (CSP) manipulees ex-vivo est une solution technique prometteuse pour une medecine regenerative. Cependant, la culture des CSP peut generer des anomalies genetiques telles que des anomalies chromosomiques ou infra-caryotypiques [1] , [2] . Notre travail vise a comprendre la nature et la cause de ces anomalies genetiques de maniere a pouvoir les limiter et ouvrir la voie a de futures applications therapeutiques reposant sur l’utilisation des CSP. Methodes Pour repertorier les anomalies genetiques publiees dans les CSP et identifier en particulier les anomalies hyper-recurrentes, nous avons cree une base de donnees « Stem cell gEnetic Abnormalities database » (SEAdb). Resultats Nous avons associe la base SEAdb a un navigateur genomique permettant de visualiser directement les anomalies genetiques sur le genome humain. Nous avons repertorie plus de 404 000 anomalies et variants d’ADN dans les CSP, issus de pres de 100 publications internationales. En integrant des filtres robustes permettant d’exclure les polymorphismes, nous avons pu determiner les anomalies les plus recurrentes observees dans les CSP. Conclusion SEAdb presente un interet majeur dans la description des anomalies genetiques dans les CSP et la comprehension des causes de leur survenue. SEAdb est une base de donnee qui sera librement accessible la fin de l’annee 2016.