Research Report
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World J Stem Cells. Apr 26, 2014; 6(2): 248-255
Published online Apr 26, 2014. doi: 10.4252/wjsc.v6.i2.248
Embryonic stem cell-derived neural progenitors as non-tumorigenic source for dopaminergic neurons
Mei-Chih Liao, Mihaela Diaconu, Sebastian Monecke, Patrick Collombat, Charles Timaeus, Tanja Kuhlmann, Walter Paulus, Claudia Trenkwalder, Ralf Dressel, Ahmed Mansouri
Mei-Chih Liao, Mihaela Diaconu, Ahmed Mansouri, Department of Molecular Cell Biology, Max-Planck-Institute for Biophysical Chemistry, 37077 Göttingen, Germany
Sebastian Monecke, Ralf Dressel, Department of Cellular and Molecular Immunology, University Medical Center Göttingen, 37073 Göttingen, Germany
Patrick Collombat, Univerité Nice Sophia Antipolis, UMR INSERM 1091/CNRS 7277/UNS, Diabetes Genetics Team, Institut de Biologie Valrose, 06100 Nice, France
Charles Timaeus, Walter Paulus, Ahmed Mansouri, Department of Clinical Neurophysiology, University Medical Center Göttingen, 37075 Göttingen, Germany
Tanja Kuhlmann, Institute of Neuropathology, University of Münster, 48149 Münster, Germany
Claudia Trenkwalder, Paracelsus-Elena-Klinik, 34128 Kassel, Germany
Patrick Collombat, Ahmed Mansouri, Genome and Stem Cell Center, GENKOK, Erciyes University, 38039 Kayseri, Turkey
Author contributions: Liao MC, Diaconu M, Monecke S, Collombat P, Timaeus C, Kuhlmann T, Dressel R and Mansouri A performed the experiments; Paulus W and Trenkwalder C contributed to the conception of the study; Dressel R and Mansouri A designed the study, wrote the manuscript and share senior authorship; all the authors approved the final version of the manuscript.
Supported by The German Federal Ministry for Education and Research (BMBF), No. 01GN0818 and No. 01GN0819; the Max-Planck Society, and initially by the Dr. Helmut Storz Stiftung
Correspondence to: Ahmed Mansouri, PhD, Professor, Department of Molecular Cell Biology, Max-Planck-Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany. amansou@gwdg.de
Telephone: +49-551-2011490 Fax: +49-551-2011504
Received: September 17, 2013
Revised: December 14, 2013
Accepted: January 13, 2014
Published online: April 26, 2014
Processing time: 222 Days and 18.8 Hours
Abstract

AIM: To find a safe source for dopaminergic neurons, we generated neural progenitor cell lines from human embryonic stem cells.

METHODS: The human embryonic stem (hES) cell line H9 was used to generate human neural progenitor (HNP) cell lines. The resulting HNP cell lines were differentiated into dopaminergic neurons and analyzed by quantitative real-time polymerase chain reaction and immunofluorescence for the expression of neuronal differentiation markers, including beta-III tubulin (TUJ1) and tyrosine hydroxylase (TH). To assess the risk of teratoma or other tumor formation, HNP cell lines and mouse neuronal progenitor (MNP) cell lines were injected subcutaneously into immunodeficient SCID/beige mice.

RESULTS: We developed a fairly simple and fast protocol to obtain HNP cell lines from hES cells. These cell lines, which can be stored in liquid nitrogen for several years, have the potential to differentiate in vitro into dopaminergic neurons. Following day 30 of differentiation culture, the majority of the cells analyzed expressed the neuronal marker TUJ1 and a high proportion of these cells were positive for TH, indicating differentiation into dopaminergic neurons. In contrast to H9 ES cells, the HNP cell lines did not form tumors in immunodeficient SCID/beige mice within 6 mo after subcutaneous injection. Similarly, no tumors developed after injection of MNP cells. Notably, mouse ES cells or neuronal cells directly differentiated from mouse ES cells formed teratomas in more than 90% of the recipients.

CONCLUSION: Our findings indicate that neural progenitor cell lines can differentiate into dopaminergic neurons and bear no risk of generating teratomas or other tumors in immunodeficient mice.

Keywords: Human embryonic stem cells; Neural progenitor cells; Teratoma; Pluripotency; Dopaminergic neurons

Core tip: The use of pluripotent cells as a source for the generation of neuronal tissue for transplantation suffers from the risk of teratoma formation. To circumvent this problem, we have developed a simple and fast protocol to obtain human neural progenitor (HNP) cell lines from embryonic stem cells. These HNP cell lines have the potential to differentiate in vitro into dopaminergic neurons. After injection into immunodeficient SCID/beige mice, they did not form tumors even after 6 mo. These findings indicate that HNP cell lines can differentiate into dopaminergic neurons and bear no risk of generating teratomas in immunodeficient mice.