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.
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.
Authors: Jonathan S Draper; Kath Smith; Paul Gokhale; Harry D Moore; Edna Maltby; Julie Johnson; Lorraine Meisner; Thomas P Zwaka; James A Thomson; Peter W Andrews Journal: Nat Biotechnol Date: 2003-12-07 Impact factor: 54.908
Authors: Chun-Ting Lee; Raphael M Bendriem; Abigail A Kindberg; Lila T Worden; Melanie P Williams; Tomas Drgon; Barbara S Mallon; Brandon K Harvey; Christopher T Richie; Rebecca S Hamilton; Jia Chen; Stacie L Errico; Shang-Yi A Tsai; George R Uhl; William J Freed Journal: Cell Rep Date: 2015-01-29 Impact factor: 9.423
Authors: Sara Lindström; Malin Eriksson; Tandis Vazin; Julia Sandberg; Joakim Lundeberg; Jonas Frisén; Helene Andersson-Svahn Journal: PLoS One Date: 2009-09-14 Impact factor: 3.240
Authors: Tandis Vazin; Kevin G Becker; Jia Chen; Charles E Spivak; Carl R Lupica; Yongqing Zhang; Lila Worden; William J Freed Journal: PLoS One Date: 2009-08-12 Impact factor: 3.240