Sample_geo_accession | GSM382206
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Sample_status | Public on Apr 11 2009
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Sample_submission_date | Mar 19 2009
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Sample_last_update_date | Apr 10 2009
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Sample_type | RNA
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Sample_channel_count | 1
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Sample_organism_ch1 | Homo sapiens
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Sample_taxid_ch1 | 9606
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Sample_biomaterial_provider_ch1 | HUES9 Kindly provided by Dr D. Melton
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Sample_treatment_protocol_ch1 | hESCs were treated with 10uM SB-431542 and described as SB-EB further at day 10 the EBs were plated and grown in 1uM SB-431542 and recalled as SB-OG.
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Sample_growth_protocol_ch1 | EB formation:
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Sample_growth_protocol_ch1 | To induce differentiation of hESCs into embryoid bodies (EB), hESC were disaggregated, using trypsin/EDTA (0.1%/ 1 mM, Gibco, 25300-0549), into small clumps containing 5-10 cells, and transferred to low-adhesion plastic Petri dishes (Costar Ultra Low Attachment; Corning Life Sciences, Acton, MA) in KO-Media without addition of bFGF and β-mercaptoethanol. However, the cells were induced with SB-431542 ([10 uM]). Embryoid bodies were formed after 3 days in culture and media change was performed at days 3, 6 and 8 by gravity sedimentation for 10 minutes at 37oC, 5% CO2.
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Sample_growth_protocol_ch1 | EB-explant outgrowth culture:
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Sample_growth_protocol_ch1 | At day 10, hEBs (SB) were transferred to fibronectin-precoated (10 µg/ml, Sigma) plates in chemically defined medium (CDM) that allowed cellular outgrowth and formation of a monolayer. CDM was composed of DMEM/F-12 (Invitrogen) supplemented with bovine serum albumin (0.5%) (Fraction V BSA, Sigma), penicillin/Streptomyocin (1%), lipids (1%) and ITS (10%) (All Invitrogen). The cultures were treated with SB-431542 ([1 uM]) at all time. The hEB-derived outgrowth cultures (OG) in the presence of SB-431542 (SB-OG) were allowed to grow to confluency; subsequent passaging was performed using trypsin/EDTA (0.05% trypsin + 0.53 mM EDTA).
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Sample_molecule_ch1 | total RNA
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Sample_extract_protocol_ch1 | Frozen samples using RNeasy Mini kit (Qiagen, Copenhagen, Denamrk) for gene expression analysis.
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Sample_label_ch1 | Biotin
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Sample_label_protocol_ch1 | GeneChip IVT Labeling Kit was used for labelling
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Sample_hyb_protocol | Eukaryotic Target Hybridization method was used according to Affymatrix protocol
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Sample_scan_protocol | GeneChip® Scanner 3000 was used according to Affymatrix protocol
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Sample_data_processing | All protocols were from Affymatrix Gene Expression Analysis Technical Manual
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Sample_platform_id | GPL570
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Sample_contact_name | Amer,,Mahmood
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Sample_contact_email | amahmood@health.sdu.dk
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Sample_contact_phone | +4529726156
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Sample_contact_laboratory | Molecular Endocrinology laboratory (KMEB)
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Sample_contact_department | Department of Endocrinology
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Sample_contact_institute | University Hospital of Odense & University of South Denmark
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Sample_contact_address | Fyrreparken 66, 2 tv
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Sample_contact_city | Odense
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Sample_contact_zip/postal_code | 5240
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Sample_contact_country | Denmark
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Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM382nnn/GSM382206/suppl/GSM382206.CEL.gz
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Sample_supplementary_file | ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM382nnn/GSM382206/suppl/GSM382206.CHP.gz
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Sample_series_id | GSE15553
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Sample_data_row_count | 54675
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