PMC:2063610 / 41324-42812 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/2063610","sourcedb":"PMC","sourceid":"2063610","source_url":"http://www.ncbi.nlm.nih.gov/pmc/2063610","text":"Human embryonic and mesenchymal stem cell culture\nThe federally approved hESC line, H7 (NIH no. WA07, obtained from WiCell Research Institue, Madison, WI, USA), was used in accordance with the UC Berkeley and UC San Francisco Committee on Human Research guidelines, and in accordance with NIH guidelines. To propagate hESCs, routine culturing and maintenance was performed using standard in vitro conditions for both feeder-dependent and feeder-free cultures (Geron Corporation, 2002). Briefly, hESCs grown on MEFs were cultured in standard hESC medium [Knockout™ DMEM, 20% KSR, 1% NEAA, 1 mm l-glutamine (Invitrogen), 0.1 mmβ-mercaptoethanol (Sigma)] and were supplemented with 4 ng mL−1 hbFGF (Invitrogen). Feeder-free hESC cultures were maintained in MEF-conditioned hESC medium (MCM), 4 ng mL−1 hbFGF. Differentiation medium for hESCs (DMEM/FBS) was made by replacing KSR with 20% FBS (Hyclone, Logan, UH, USA). hMSCs were maintained in mesenchymal stem cell GM, MSC-GM™ and were cultured according to supplier recommendations (Cambrex Walkersville, MD, USA). hESCs and hMSCs were typically seeded onto chambered slides coated with a 3% GFR Matrigel™ (BD Biosciences) substrate in PBS. Cells were typically incubated for 48 h at 37 °C, 5% CO2, under the various experimental conditions employed, then were fixed with 70% EtOH/PBS at 4 °C. hESCs and hMSCs were analyzed 24–48 h after experimental treatments, during which no apoptosis-related differences in cell numbers were observed.","divisions":[{"label":"Title","span":{"begin":0,"end":49}}],"tracks":[{"project":"CellFinder","denotations":[{"id":"T555","span":{"begin":833,"end":838},"obj":"CellType"},{"id":"T556","span":{"begin":1343,"end":1348},"obj":"CellType"},{"id":"T557","span":{"begin":96,"end":100},"obj":"CellLine"},{"id":"T558","span":{"begin":495,"end":500},"obj":"CellType"},{"id":"T559","span":{"begin":510,"end":514},"obj":"CellType"},{"id":"T560","span":{"begin":318,"end":323},"obj":"CellType"},{"id":"T561","span":{"begin":709,"end":715},"obj":"CellType"},{"id":"T562","span":{"begin":689,"end":694},"obj":"GeneProtein"},{"id":"T563","span":{"begin":438,"end":444},"obj":"CellType"},{"id":"T564","span":{"begin":1353,"end":1358},"obj":"CellType"},{"id":"T565","span":{"begin":20,"end":31},"obj":"Anatomy"},{"id":"T566","span":{"begin":6,"end":15},"obj":"Anatomy"},{"id":"T567","span":{"begin":1064,"end":1069},"obj":"CellType"},{"id":"T568","span":{"begin":941,"end":962},"obj":"CellType"},{"id":"T569","span":{"begin":916,"end":921},"obj":"CellType"},{"id":"T570","span":{"begin":1074,"end":1079},"obj":"CellType"},{"id":"T571","span":{"begin":417,"end":423},"obj":"CellType"},{"id":"T572","span":{"begin":799,"end":804},"obj":"GeneProtein"},{"id":"T573","span":{"begin":941,"end":952},"obj":"Anatomy"},{"id":"T574","span":{"begin":20,"end":41},"obj":"CellType"},{"id":"T575","span":{"begin":84,"end":86},"obj":"CellLine"}],"attributes":[{"subj":"T555","pred":"source","obj":"CellFinder"},{"subj":"T556","pred":"source","obj":"CellFinder"},{"subj":"T557","pred":"source","obj":"CellFinder"},{"subj":"T558","pred":"source","obj":"CellFinder"},{"subj":"T559","pred":"source","obj":"CellFinder"},{"subj":"T560","pred":"source","obj":"CellFinder"},{"subj":"T561","pred":"source","obj":"CellFinder"},{"subj":"T562","pred":"source","obj":"CellFinder"},{"subj":"T563","pred":"source","obj":"CellFinder"},{"subj":"T564","pred":"source","obj":"CellFinder"},{"subj":"T565","pred":"source","obj":"CellFinder"},{"subj":"T566","pred":"source","obj":"CellFinder"},{"subj":"T567","pred":"source","obj":"CellFinder"},{"subj":"T568","pred":"source","obj":"CellFinder"},{"subj":"T569","pred":"source","obj":"CellFinder"},{"subj":"T570","pred":"source","obj":"CellFinder"},{"subj":"T571","pred":"source","obj":"CellFinder"},{"subj":"T572","pred":"source","obj":"CellFinder"},{"subj":"T573","pred":"source","obj":"CellFinder"},{"subj":"T574","pred":"source","obj":"CellFinder"},{"subj":"T575","pred":"source","obj":"CellFinder"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"CellFinder","color":"#9395ec","default":true}]}]}}