Dopaminergic neurons derived from BG01V2, a variant of human embryonic stem cell line BG01

Article type: Research Article

Authors: Vazin, Tandis^; | Chen, Jia | Spivak, Charles E. | Amable, Rose | Gabitzsch, Emily | Lee, Chun-Ting | Lupica, Carl R. | Freed, William J.

Affiliations: Development and Plasticity Section, Cellular Neurobiology Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Department of Health and Human Services, Baltimore, MD, USA | Cellular Neurophysiology Section, Cellular Neurobiology Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Department of Health and Human Services, Baltimore, MD, USA | School of Biotechnology, Department of Gene Technology, KTH, Royal Institute of Technology, AlbaNova University Center, Stockholm, Sweden

Note: [] Corresponding author: Tandis Vazin, NIDA Intramural Research Program, Cellular Neurobiology Research Branch, 333 Cassell Drive, Triad Bldg., Room 3303, Baltimore, MD 21224, USA. Tel.: +1 443 740 6137; Fax: +1 443 740 2123; E-mail: vazint@mail.nih.gov

Abstract: Background and Purpose: Human embryonic stem cells (hESC) are considered a renewable source of dopamine producing neurons, and are of particular interest for their potential clinical use in Parkinson's disease. In this study, we characterized human dopaminergic neurons generated by stromal-derived inducing activity (SDIA) from BG01V2, a strain of human embryonic stem cell line, BG01, characterized by a chromosome 17 trisomy. Similar chromosomal changes have been repeatedly observed in hESC cultures in different laboratories, indicating the importance of chromosome 17 for growth and adaptation of hESC to culture. Methods: We investigated in vitro proliferation of differentiating cells using a BrDU incorporation assay, and monitored the cell population in long term cultures. Despite the cytogenetic abnormality, TH+ neurons were postmitotic at all stages of differentiation. After 30 days of differentiation, cell division ceased in 91% of the overall population of cells in the culture, indicating intact cell cycle regulation. Results: Expression of midbrain specific marker genes (Otx2, Pax5, Msx-1) showed differentiation of hESC-derived neural progenitor cells into midbrain specific dopamine neurons. These neurons expressed the dopamine transporter (DAT), and displayed functional DAT activity and electrical excitability. Conclusions: TH+ cells derived from the BG01V2 hESC line using SDIA are postmitotic and have functional characteristics of normal dopaminergic neurons.

Keywords: SDIA, embryonic stem cells, human, dopaminergic neuron, PA6 cells, BG01V2, trisomy 17

Journal: Restorative Neurology and Neuroscience, vol. 26, no. 6, pp. 447-458, 2008