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Transplantable Neural Progenitor Populations Derived from Rhesus Monkey Embryonic Stem Cells

^a Department of Reproduction and Development, Kunming Institute of Zoology, The Chinese Academy of Sciences, Kunming, Yunnan, China;
^b Kunming Primate Research Center, The Chinese Academy of Sciences, Kunming, Yunnan, China;
^c Graduate School, The Chinese Academy of Sciences, Beijing, China;
^d Section of Cognitive Brain Research, Kunming Institute of Zoology, The Chinese Academy of Sciences, Kunming, Yunnan, China;
^e Oregon National Primate Research Center, Portland, Oregon, USA;
^f Institute of Zoology, The Chinese Academy of Sciences, Beijing, China

Key Words. Rhesus monkey embryonic stem cells • Neural progenitors • Hepatocyte growth factor • G5 supplement • Differentiation

Correspondence: Weizhi Ji, Ph.D., Kunming Primate Research Center and Kunming Institute of Zoology, The Chinese Academy of Sciences, 32 Jiaochang Donglu, Kunming, Yunnan, 650223, China. Telephone: 86-871-5139413; Fax: 86-871-5139413; e-mail: wji{at}mail.kiz.ac.cn; and Qi Zhou, Institute of Zoology, The Chinese Academy of Sciences, Beijing 100086, China. Telephone: 86-10-62650042; e-mail: qzhou{at}ioz.ac.cn

Cell-based therapies using embryonic stem cells (ESCs) in the treatment of neural disease will require the generation of homogenous donor neural progenitor (NP) populations. Here we describe an efficient culture system containing hepatocyte growth factor (HGF) and G5 supplement for the production of highly enriched (88.3% ± 8.1%) populations of NPs from rhesus monkey ESCs. Additional purification resulted in NP preparations that were 98% nestin positive. Moreover, NPs, as monolayers or neurospheres, could be maintained for prolonged periods of time in media containing HGF+G5 or G5 alone. In vitro differentiation and in vivo transplantation assays showed that NPs could differentiate into neurons, astrocytes, and oligodendrocytes. The kinds and quantities of differentiated cells derived from NPs were closely correlated with their niches in vivo. Glial differentiation was predominant in periventricular areas, whereas cells migrating into the cortex were mostly neurons. Cell counts showed that 2 months after transplantation, approximately 25% of transplanted NPs survived and 65%–80% of the surviving transplanted cells migrated along the ventricular wall or in a radial fashion. Subcloning demonstrated that several clonal lines derived from NPs expressed nestin and differentiated into three neural lineages in vitro and in rat brains in vivo. In contrast, some subcloned lines showed restricted differentiation both in vitro and in vivo in rat brains. These observations set the stage for obtaining highly enriched NPs and evaluating the efficacy of NP-based transplantation therapy in the nonhuman primate and will provide a platform for probing the molecular mechanisms that control neural induction.

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