HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints/Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Yue, F. Articles by Sasaki, K. Search for Related Content PubMed PubMed Citation Articles by Yue, F. Articles by Sasaki, K.

EMBRYONIC STEM CELLS

Induction of Midbrain Dopaminergic Neurons from Primate Embryonic Stem Cells by Coculture with Sertoli Cells

Department of Anatomy and Organ Technology, Institute of Organ Transplants, Reconstructive Medicine and Tissue Engineering, Shinshu University Graduate School of Medicine, Asahi, Matsumoto, Nagano, Japan

Key Words. Embryonic stem cells • Primate • Dopaminergic neuron • Differentiation • Sertoli cells • Coculture

Correspondence: Fengming Yue, Ph.D., Department of Anatomy and Organ Technology, Institute of Organ Transplants, Reconstructive Medicine and Tissue Engineering, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Nagano, Japan Telephone: 81-263-372590; Fax: 81-263-373093; e-mail: yuefm{at}sch.md.shinshu-u.ac.jp

Received August 24, 2005; accepted for publication April 5, 2006.

The aim of this study was to produce dopaminergic neurons from primate embryonic stem (ES) cells following coculture with mouse Sertoli cells. After 3 weeks of induction, immunostaining revealed that 90% ± 9% of the colonies contained tyrosine hydroxylase-positive (TH⁺) neurons, and 60% ± 7% of the tubulin ß III-positive (Tuj III⁺) neurons were TH⁺. Reverse transcription-polymerase chain reaction analyses showed that Sertoli-induced neurons expressed midbrain dopaminergic neuron markers, including TH, dopamine transporter, aromatic amino acid decarboxylase (AADC), receptors such as TrkB and TrkC, and transcription factors NurrI and Lmx1b. Neurons that had been differentiated on Sertoli cells were positive for Pax2, En1, and AADC, midbrain-related markers, and negative for dopamine-ß-hydroxylase, a marker of noradrenergic neurons. These Sertoli cell-induced dopaminergic cells can release dopamine when depolarized by high K⁺. Sertoli cell-conditioned medium contained glial cell line-derived neurotrophic factor (GDNF) and supported neuronal differentiation. After pretreatment with anti-GDNF antibody, the percentage of Tuj III⁺ colonies was reduced to 14%. Thus, GDNF contributed significantly to inducing primate ES cells into dopaminergic neurons. When transplanted into a 6-hydroxydopamine-treated Parkinson’s disease model, primate-derived dopaminergic neurons integrated into the mouse striatum. Two weeks after transplantation, surviving TH⁺ cells were present. These TH⁺ cells survived for 2 months. Therefore, the induction method of coculture ES cells with Sertoli cells provides an unlimited source of primate cells for the study of pathogenesis and transplantation in Parkinson’s disease.

This article has been cited by other articles:

				Z. He, J. Jiang, M.-C. Hofmann, and M. Dym Gfra1 Silencing in Mouse Spermatogonial Stem Cells Results in Their Differentiation Via the Inactivation of RET Tyrosine Kinase Biol Reprod, October 1, 2007; 77(4): 723 - 733. [Abstract] [Full Text] [PDF]