Integration-deficient lentivectors: an effective strategy to purify and differentiate human embryonic stem cell-derived hepatic progenitors.

Guanghua Yang 1 Karim Si-Tayeb 1 Sébastien Corbineau 1 Rémi Vernet 1 Régis Gayon 2 Noushin Dianat 1 Clémence Martinet 1 Denis Clay 1 Sylvie Goulinet-Mainot 1 Gérard Tachdjian 3 Deborah Burks 4 Ludovic Vallier 5 Pascale Bouillé 2 Anne Dubart-Kupperschmitt 1, * Anne Weber 1
* Auteur correspondant
1 Les cellules souches : de leurs niches à leurs applications thérapeutiques
2 Vectalys SAS
3 Service d'histologie, embryologie et cytogénétique [Béclère]
4 CIBER de Diabetes y Obesidad
5 Department of Surgery
Abstract : BACKGROUND: Human pluripotent stem cells (hPSCs) hold great promise for applications in regenerative medicine. However, the safety of cell therapy using differentiated hPSC derivatives must be improved through methods that will permit the transplantation of homogenous populations of a specific cell type. To date, purification of progenitors and mature cells generated from either embryonic or induced pluripotent stem cells remains challenging with use of conventional methods. RESULTS: We used lentivectors encoding green fluorescent protein (GFP) driven by the liver-specific apoliprotein A-II (APOA-II) promoter to purify human hepatic progenitors. We evaluated both integrating and integration-defective lentivectors in combination with an HIV integrase inhibitor. A human embryonic stem cell line was differentiated into hepatic progenitors using a chemically defined protocol. Subsequently, cells were transduced and sorted at day 16 of differentiation to obtain a cell population enriched in hepatic progenitor cells. After sorting, more than 99% of these APOA-II-GFP-positive cells expressed hepatoblast markers such as α-fetoprotein and cytokeratin 19. When further cultured for 16 days, these cells underwent differentiation into more mature cells and exhibited hepatocyte properties such as albumin secretion. Moreover, they were devoid of vector DNA integration. CONCLUSIONS: We have developed an effective strategy to purify human hepatic cells from cultures of differentiating hPSCs, producing a novel tool that could be used not only for cell therapy but also for in vitro applications such as drug screening. The present strategy should also be suitable for the purification of a broad range of cell types derived from either pluripotent or adult stem cells.
Keywords : Tissue-specific progenitor cells Hepatoblasts Purification Non-integrating lentiviral vectors
Type de document :
Article dans une revue
BMC Biology, BioMed Central, 2013, 11 (1), pp.86. 〈10.1186/1741-7007-11-86〉
Liste complète des métadonnées
http://www.hal.inserm.fr/inserm-00852086
Contributeur : Ed. Bmc <>
Soumis le : mardi 20 août 2013 - 06:04:29
Dernière modification le : mercredi 29 novembre 2017 - 10:29:52
Document(s) archivé(s) le : jeudi 21 novembre 2013 - 04:13:24

Fichiers

1741-7007-11-86.pdf
Fichiers éditeurs autorisés sur une archive ouverte

Identifiants

Collections

Citation

Guanghua Yang, Karim Si-Tayeb, Sébastien Corbineau, Rémi Vernet, Régis Gayon, et al.. Integration-deficient lentivectors: an effective strategy to purify and differentiate human embryonic stem cell-derived hepatic progenitors.. BMC Biology, BioMed Central, 2013, 11 (1), pp.86. 〈10.1186/1741-7007-11-86〉. 〈inserm-00852086〉

Exporter

BibTeX TEI DC DCterms EndNote

Partager

Métriques

Consultations de la notice

430

Téléchargements de fichiers

262

Contact

support.ccsd.cnrs.fr
hal.support@ccsd.cnrs.fr
Logo CCSD