PMC:1802744 / 25941-27515 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/1802744","sourcedb":"PMC","sourceid":"1802744","source_url":"http://www.ncbi.nlm.nih.gov/pmc/1802744","text":"Commitment of hESCs towards neuronal/dopaminergic differentiation pathway\nCo-culturing with the PA6 stromal cell line (MC3T3-G2/Pa6, from RIKEN Cell Bank Japan (RCB 1127), derived from newborn mouse calvaria rapidly generates high numbers of DA neurons from mouse and monkey ESCs by an unknown mechanism named stromal-derived inducing activity (SDIA; [28,29]). For differentiation experiments, PA6 cells were plated on gelatine-coated T25 flasks at 16 × 103 cells/cm2 (400,000 cells/flask) density 2 days prior to introducing hESCs into the co-culture and cultured at PA6 culturing media (containing minimum essential medium alpha (α-MEM) supplemented with 10% FCS and 0.5% Penicillin/Streptomycin). Alternatively, PA6 cells were plated over Type I collagen-coated glass cover-slips placed in wells of 4-well-plates (50,000 cells/well, for immunocytochemical (ICC) analysis). Three hours prior to initiation of co-culture, PA6 cells were rinsed 3 times with PBS and media was replaced with co-culture media (Glasgow's modified Eagle's media (G-MEM) supplemented with 8% knock-out serum replacement (KSR), 2 mM glutamine, 0.1 mM non-essential amino-acids (NEAA), 1 mM pyruvate, 0.1 mM β-mercaptoethanol (βME) and 4 ng/μl bFGF). Fragments of hESC colonies (80–90 per flask; 4–5 per well of 4-well-plate) presenting undifferentiated morphology were manually passaged onto the confluent PA6 monolayer and cell co-cultures were maintained at 37°C, 5% CO2, 95% humidity settings. One half of the co-culture medium was replaced every other day for first 10 days, and daily onwards.","divisions":[{"label":"Title","span":{"begin":0,"end":73}}],"tracks":[{"project":"CellFinder","denotations":[{"id":"T342","span":{"begin":742,"end":757},"obj":"GeneProtein"},{"id":"T343","span":{"begin":268,"end":274},"obj":"Species"},{"id":"T344","span":{"begin":258,"end":263},"obj":"Species"},{"id":"T345","span":{"begin":1220,"end":1224},"obj":"GeneProtein"},{"id":"T346","span":{"begin":1383,"end":1386},"obj":"CellLine"},{"id":"T347","span":{"begin":394,"end":403},"obj":"CellType"},{"id":"T348","span":{"begin":96,"end":99},"obj":"CellLine"},{"id":"T349","span":{"begin":923,"end":926},"obj":"CellLine"},{"id":"T350","span":{"begin":28,"end":36},"obj":"Anatomy"},{"id":"T351","span":{"begin":242,"end":251},"obj":"CellType"},{"id":"T352","span":{"begin":568,"end":571},"obj":"CellLine"},{"id":"T353","span":{"begin":28,"end":36},"obj":"CellType"},{"id":"T354","span":{"begin":345,"end":349},"obj":"GeneProtein"},{"id":"T355","span":{"begin":193,"end":198},"obj":"Species"},{"id":"T356","span":{"begin":923,"end":932},"obj":"CellType"},{"id":"T357","span":{"begin":394,"end":397},"obj":"CellLine"},{"id":"T358","span":{"begin":199,"end":207},"obj":"Anatomy"},{"id":"T359","span":{"begin":245,"end":252},"obj":"CellType"},{"id":"T360","span":{"begin":310,"end":343},"obj":"GeneProtein"},{"id":"T361","span":{"begin":526,"end":531},"obj":"CellType"},{"id":"T362","span":{"begin":14,"end":19},"obj":"CellType"},{"id":"T363","span":{"begin":715,"end":718},"obj":"CellLine"},{"id":"T364","span":{"begin":715,"end":724},"obj":"CellType"},{"id":"T365","span":{"begin":275,"end":279},"obj":"CellType"}],"attributes":[{"subj":"T342","pred":"source","obj":"CellFinder"},{"subj":"T343","pred":"source","obj":"CellFinder"},{"subj":"T344","pred":"source","obj":"CellFinder"},{"subj":"T345","pred":"source","obj":"CellFinder"},{"subj":"T346","pred":"source","obj":"CellFinder"},{"subj":"T347","pred":"source","obj":"CellFinder"},{"subj":"T348","pred":"source","obj":"CellFinder"},{"subj":"T349","pred":"source","obj":"CellFinder"},{"subj":"T350","pred":"source","obj":"CellFinder"},{"subj":"T351","pred":"source","obj":"CellFinder"},{"subj":"T352","pred":"source","obj":"CellFinder"},{"subj":"T353","pred":"source","obj":"CellFinder"},{"subj":"T354","pred":"source","obj":"CellFinder"},{"subj":"T355","pred":"source","obj":"CellFinder"},{"subj":"T356","pred":"source","obj":"CellFinder"},{"subj":"T357","pred":"source","obj":"CellFinder"},{"subj":"T358","pred":"source","obj":"CellFinder"},{"subj":"T359","pred":"source","obj":"CellFinder"},{"subj":"T360","pred":"source","obj":"CellFinder"},{"subj":"T361","pred":"source","obj":"CellFinder"},{"subj":"T362","pred":"source","obj":"CellFinder"},{"subj":"T363","pred":"source","obj":"CellFinder"},{"subj":"T364","pred":"source","obj":"CellFinder"},{"subj":"T365","pred":"source","obj":"CellFinder"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"CellFinder","color":"#ec93e3","default":true}]}]}}