Assessment of Stromal-Derived Inducing Activity in the Generation of Dopaminergic Neurons from Human Embryonic Stem Cells

@article{citeulike:2643013, abstract = {Producing dopaminergic (DA) neurons is a major goal of human embryonic stem cell (hESC) research. DA neurons can be differentiated from hESC by co-culture with the mouse PA6 stromal cell line; this differentiation-inducing effect is termed stromal-derived inducing activity (SDIA). The molecular and biochemical nature of SDIA is, however, unknown. Various studies have suggested that SDIA involves either a fixation-resistant component located on the PA6 cell surface, or factors secreted into the medium by PA6 cells. To address this question, hESC were co-cultured with PA6 cells for 12 days, then further differentiated with SHH, FGF-8 and glial cell line-derived neurotrophic factor. After 18 days, 34\% of cells were tyrosine hydroxylase (TH)+. When PA6 cells were fixed or irradiated, the number of TH+ cells was decreased by 3-fold, while mitomycin-c treatment of feeder cells decreased the number of TH+ cells by 32\%. The neural-inducing effect of PA6 cells, as monitored by {beta}-III-tubulin expression, was minimally affected by mitomycin-c treatment or fixation, but was decreased 50\% by irradiation. Medium conditioned by PA6 cells was ineffective in differentiating TH+ cells when used alone. Conditioned medium combined with heparin and/or fixed PA6 cells produced TH+ cell differentiation, although less effectively than PA6 cell co-culture. Thus, PA6 cell surface activity is required for neural differentiation of hESC, but secreted factors are required for the specific DA neuron-inducing effect. 10.1634/stemcells.2008-0039}, author = {Vazin, Tandis and Chen, Jia and Lee, Chun T. and Amable, Rose and Freed, William J. }, citeulike-article-id = {2643013}, doi = {http://dx.doi.org/10.1634/stemcells.2008-0039}, journal = {Stem Cells}, keywords = {cell, dopamine, neuron, pa6, stem}, month = {April}, pages = {2008-0039+}, posted-at = {2008-04-08 23:02:07}, priority = {2}, title = {Assessment of Stromal-Derived Inducing Activity in the Generation of Dopaminergic Neurons from Human Embryonic Stem Cells}, url = {http://dx.doi.org/10.1634/stemcells.2008-0039}, year = {2008} }

TY - JOUR ID - citeulike:2643013 L3 - citeulike-article-id:2643013 N2 - Producing dopaminergic (DA) neurons is a major goal of human embryonic stem cell (hESC) research. DA neurons can be differentiated from hESC by co-culture with the mouse PA6 stromal cell line; this differentiation-inducing effect is termed stromal-derived inducing activity (SDIA). The molecular and biochemical nature of SDIA is, however, unknown. Various studies have suggested that SDIA involves either a fixation-resistant component located on the PA6 cell surface, or factors secreted into the medium by PA6 cells. To address this question, hESC were co-cultured with PA6 cells for 12 days, then further differentiated with SHH, FGF-8 and glial cell line-derived neurotrophic factor. After 18 days, 34% of cells were tyrosine hydroxylase (TH)+. When PA6 cells were fixed or irradiated, the number of TH+ cells was decreased by 3-fold, while mitomycin-c treatment of feeder cells decreased the number of TH+ cells by 32%. The neural-inducing effect of PA6 cells, as monitored by {beta}-III-tubulin expression, was minimally affected by mitomycin-c treatment or fixation, but was decreased 50% by irradiation. Medium conditioned by PA6 cells was ineffective in differentiating TH+ cells when used alone. Conditioned medium combined with heparin and/or fixed PA6 cells produced TH+ cell differentiation, although less effectively than PA6 cell co-culture. Thus, PA6 cell surface activity is required for neural differentiation of hESC, but secreted factors are required for the specific DA neuron-inducing effect. 10.1634/stemcells.2008-0039 JF - Stem Cells TI - Assessment of Stromal-Derived Inducing Activity in the Generation of Dopaminergic Neurons from Human Embryonic Stem Cells SP - 2008-0039 KW - cell KW - dopamine KW - neuron KW - pa6 KW - stem AU - Vazin, Tandis AU - Chen, Jia AU - Lee, Chun AU - Amable, Rose AU - Freed, William PY - 2008/04/03/ UR - http://dx.doi.org/10.1634/stemcells.2008-0039 ER -