Result

Search results for the GEO ID: GSE14503

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Description

Characteristics

GSM239824

GSM239825

GSM239826

GSM239827

GSM239828

GPL570

hES-T3 derived embryoid bodies, biological rep2

The T3EB embryoid bodies were formed in 7 days from hES-T3 by mechanically dissection , and they resembled early embryo development to some extent

gender: female passage: 36 sample type: embryoid bodies derived from T3ES

The mRNA profiles from the undifferentiated human embryonic stem cell line hES-T3 (T3ES), hES-T3 derived embryoid bodies (T3EB) and hES-T3 differentiated fibroblast-like cells (T3DF) were quantitatively determined.

Sample_geo_accession	GSM239828
Sample_status	Public on Oct 25 2008
Sample_submission_date	Oct 26 2007
Sample_last_update_date	Aug 14 2011
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Homo sapiens
Sample_taxid_ch1	9606
Sample_treatment_protocol_ch1	Non
Sample_growth_protocol_ch1	The formation of embryoid bodies (EBs) was induced by mechanically dissecting undifferentiated hES-T3 colonies into pieces that were transferred and grown with very slow shaking (20 rpm) in the EB medium containing 80% knockout DMEM, 20% ES-qualified FBS (GIBCO), 1% non-essential amino acids, and 1 mM L-glutamine without MEF feeder layer in bacterial Petri dish plate(£\-plus, 10 cm) under 5% CO2 at 37oC for 7 days with change of medium every two days. These embryoid bodies are designated as T3EB.
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	Trizol extraction of total RNA was performed according to the manufacturer's instructions. Total RNAs extracted using TRIZOL reagent were quantified spectrophotometrically. The cDNA synthesis was carried out by using SUPERSCRIPTTM one-step RT-PCR kit (Invitrogen). For the reverse transcription step, the whole 5 ul of the resuspended RNAs were incubated for 60 min at 42oC, then 15 min at 72oC in 50 ul of reaction mixture containing 25 ul of 2x Reaction Mix (Invitrogen) and 1 ul of RT/PLANTINUM Taq Mix (Invitrogen). 28.5 ul of the cDNAs present in the 50 ul RT reaction mixture purified by Microarray Target Purification Kit (Roche Applied Science, Indianapolis, ID, USA, http://www.roche-applies-science.com) were used as templates to amplify cDNA by Microarray Target Amplification Kit. The amplified cDNAs were purified by Microarray Target Purification Kit, and complementary RNAs (cRNAs) were synthesized from 200 ng cDNA with Microarray RNA Target Synthesis Kit (Roche Applied Science).
Sample_label_ch1	biotin
Sample_label_protocol_ch1	Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5 micrograms purified cRNA (Expression Analysis Technical Manual, 2004, Affymetrix).
Sample_hyb_protocol	Following fragmentation, 10 micrograms of cRNA were hybridized for 16 hr at 45oC on Affymetrix Human Genome U133 plus 2.0 GeneChip. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
Sample_scan_protocol	GeneChips from the hybridization experiments were read by the Affymetrix GeneChip scanner 3000.
Sample_data_processing	The data were processed by GeneSpring software version 7.2 GC-RMA preprocessor to use on the CEL files
Sample_platform_id	GPL570
Sample_contact_name	Sung-Liang,,Yu
Sample_contact_email	slyu@ntu.edu.tw
Sample_contact_phone	886-2-23958341
Sample_contact_fax	886-2-23958341
Sample_contact_laboratory	Microarray Core Facility
Sample_contact_department	Clinical Laboratory Sciences and Medical Biotechnology
Sample_contact_institute	National Taiwan University
Sample_contact_address	Jen Ai Road Section1
Sample_contact_city	Taipei
Sample_contact_zip/postal_code	100
Sample_contact_country	Taiwan
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM239nnn/GSM239828/suppl/GSM239828.CEL.gz
Sample_series_id	GSE9440
Sample_series_id	GSE14503
Sample_data_row_count	54675

GSM362248

GPL570

Human pancreatic islet-like cell clusters, biological rep1

Human pancreatic islet-like cell clusters derived from human embryonic stem cell line hES-T3 cells

gender: female sample type: pancreatic islet-like cell clusters derived from T3ES

The mRNA profilings of T3ES, T3EB and T3pi cells were analyzed using Affymetrix Human Genome U133 plus 2.0 GeneChip according to the Manufacturer¡¦s protocols (Santa Clara, CA, USA, http://www.affymetrix.com).

Sample_geo_accession	GSM362248
Sample_status	Public on Sep 30 2010
Sample_submission_date	Jan 21 2009
Sample_last_update_date	Sep 30 2010
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Homo sapiens
Sample_taxid_ch1	9606
Sample_treatment_protocol_ch1	Non
Sample_growth_protocol_ch1	Human embryonic stem cell line hES-T3 has been continuously cultured on mitomycin C (10 ug/ml) mitotically inactivated MEF feeder in hES medium under 5% CO2 at 37oC. The hES culture medium consisted of DMEM/F12 (1:1, GIBCO) supplemented with 20% KSR (Invitrogen), 1% non-essential amino acids, 1 mM L-glutamine, 0.1 mM £]-mercaptoethanol, and 4 ng/ml human basic fibroblast growth factor (bFGF; Life Technologies). The five-stage procedure for in vitro differentiation of hES-T3 cells into pancreatic islet-like cell clusters was described as follows: Stage I: Expansion of undifferentiated hES cells. The hES-T3 cells (passage 36) were maintained on MEF feeder in hES medium containing 4 ng/ml bFGF as described above. The hES-T3 cells grown on MEF feeder were designated as T3ES. Stage II: Formation of embryoid bodies. The undifferentiated hES-T3 colonies were mechanically dissected into pieces that were transferred and grown with very slow shaking (20 rpm) in the EB medium containing 80% knockout DMEM, 20% ES-qualified FBS (GIBCO), 1% non-essential amino acids, and 1 mM L-glutamine without MEF feeder layer in bacterial Petri dish plate (£\-plus, 10 cm) under 5% CO2 at 37oC for 7 days with change of medium every two days. These embryoid bodies are designated as T3EB. Stage III: Plating EBs in ITSF medium. The EBs were transferred to 0.1% gelatin coated-plate in DMEM/F12 medium supplemented with 1% ITS (1 g/L insulin, 0.55 g/L transferring, 0.67 mg/L selenium), 5 ug/ml fibronectin and 1 mM L-glutamine under 5% CO2 at 37oC for 7 days with change of medium every two days. Stage IV: Expansion of pancreatic progenitor cells. The cells were dissociated with 0.05% trypsin at 37oC for 5 minutes, and plated on 0.1% gelatin-coated dish in DMEM/F12 medium supplemented with 1% N2 supplement (500 ug/ml insulin, 10 mg/ml transferrin, 0.63 ug/ml progesterone, 1.611 mg/ml putrascin, and 0.52 ug/ml selenite, GIBCO), 2% B27 supplement (GIBCO), 1 mM L-glutamine and 10 ng/ml bFGF under 5% CO2 at 37oC for 7 days with change of medium daily. Stage V: Formation of pancreatic islet-like cell clusters. The DMEM (containing no glucose)/F12 medium was supplemented 1% N2 supplement, 2% B27 supplement, 1mM L-glutamin, and 10 mM nicotinamide (instead of bFGF) under 5% CO2 at 37oC for 7 days with change of medium every two days. These hES-T3 derived pancreatic islet-like cell clusters were designated as T3pi.
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	Trizol extraction of total RNA was performed according to the manufacturer's instructions. Total RNAs extracted using TRIZOL reagent were quantified spectrophotometrically. The cDNA synthesis was carried out by using SUPERSCRIPTTM one-step RT-PCR kit (Invitrogen). For the reverse transcription step, the whole 5 ul of the resuspended RNAs were incubated for 60 min at 42oC, then 15 min at 72oC in 50 ul of reaction mixture containing 25 ul of 2x Reaction Mix (Invitrogen) and 1 ul of RT/PLANTINUM Taq Mix (Invitrogen). 28.5 ul of the cDNAs present in the 50 ul RT reaction mixture purified by Microarray Target Purification Kit (Roche Applied Science, Indianapolis, ID, USA, http://www.roche-applies-science.com) were used as templates to amplify cDNA by Microarray Target Amplification Kit. The amplified cDNAs were purified by Microarray Target Purification Kit, and complementary RNAs (cRNAs) were synthesized from 200 ng cDNA with Microarray RNA Target Synthesis Kit (Roche Applied Science).
Sample_label_ch1	biotin
Sample_label_protocol_ch1	Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5 micrograms purified cRNA (Expression Analysis Technical Manual, 2004, Affymetrix, http://www.affymetrix.com).
Sample_hyb_protocol	Following fragmentation, 10 micrograms of cRNA were hybridized for 16 hr at 45oC on Affymetrix Human Genome U133 plus 2.0 GeneChip. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
Sample_scan_protocol	GeneChips from the hybridization experiments were read by the Affymetrix GeneChip scanner 3000.
Sample_data_processing	The data were processed by GeneSpring GX software version 7.3.1 GC-RMA preprocessor to use on the CEL files
Sample_platform_id	GPL570
Sample_contact_name	Sung-Liang,,Yu
Sample_contact_email	slyu@ntu.edu.tw
Sample_contact_phone	886-2-23958341
Sample_contact_fax	886-2-23958341
Sample_contact_laboratory	Microarray Core Facility
Sample_contact_department	Clinical Laboratory Sciences and Medical Biotechnology
Sample_contact_institute	National Taiwan University
Sample_contact_address	Jen Ai Road Section1
Sample_contact_city	Taipei
Sample_contact_zip/postal_code	100
Sample_contact_country	Taiwan
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM362nnn/GSM362248/suppl/GSM362248.CEL.gz
Sample_series_id	GSE14503
Sample_data_row_count	54675

GSM362249

GPL570

Human pancreatic islet-like cell clusters, biological rep2

Human pancreatic islet-like cell clusters derived from human embryonic stem cell line hES-T3 cells

gender: female sample type: pancreatic islet-like cell clusters derived from T3ES

Sample_geo_accession	GSM362249
Sample_status	Public on Sep 30 2010
Sample_submission_date	Jan 21 2009
Sample_last_update_date	Sep 30 2010
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Homo sapiens
Sample_taxid_ch1	9606
Sample_treatment_protocol_ch1	Non
Sample_growth_protocol_ch1	Human embryonic stem cell line hES-T3 has been continuously cultured on mitomycin C (10 ug/ml) mitotically inactivated MEF feeder in hES medium under 5% CO2 at 37oC. The hES culture medium consisted of DMEM/F12 (1:1, GIBCO) supplemented with 20% KSR (Invitrogen), 1% non-essential amino acids, 1 mM L-glutamine, 0.1 mM £]-mercaptoethanol, and 4 ng/ml human basic fibroblast growth factor (bFGF; Life Technologies). The five-stage procedure for in vitro differentiation of hES-T3 cells into pancreatic islet-like cell clusters was described as follows: Stage I: Expansion of undifferentiated hES cells. The hES-T3 cells (passage 36) were maintained on MEF feeder in hES medium containing 4 ng/ml bFGF as described above. The hES-T3 cells grown on MEF feeder were designated as T3ES. Stage II: Formation of embryoid bodies. The undifferentiated hES-T3 colonies were mechanically dissected into pieces that were transferred and grown with very slow shaking (20 rpm) in the EB medium containing 80% knockout DMEM, 20% ES-qualified FBS (GIBCO), 1% non-essential amino acids, and 1 mM L-glutamine without MEF feeder layer in bacterial Petri dish plate (£\-plus, 10 cm) under 5% CO2 at 37oC for 7 days with change of medium every two days. These embryoid bodies are designated as T3EB. Stage III: Plating EBs in ITSF medium. The EBs were transferred to 0.1% gelatin coated-plate in DMEM/F12 medium supplemented with 1% ITS (1 g/L insulin, 0.55 g/L transferring, 0.67 mg/L selenium), 5 ug/ml fibronectin and 1 mM L-glutamine under 5% CO2 at 37oC for 7 days with change of medium every two days. Stage IV: Expansion of pancreatic progenitor cells. The cells were dissociated with 0.05% trypsin at 37oC for 5 minutes, and plated on 0.1% gelatin-coated dish in DMEM/F12 medium supplemented with 1% N2 supplement (500 ug/ml insulin, 10 mg/ml transferrin, 0.63 ug/ml progesterone, 1.611 mg/ml putrascin, and 0.52 ug/ml selenite, GIBCO), 2% B27 supplement (GIBCO), 1 mM L-glutamine and 10 ng/ml bFGF under 5% CO2 at 37oC for 7 days with change of medium daily. Stage V: Formation of pancreatic islet-like cell clusters. The DMEM (containing no glucose)/F12 medium was supplemented 1% N2 supplement, 2% B27 supplement, 1mM L-glutamin, and 10 mM nicotinamide (instead of bFGF) under 5% CO2 at 37oC for 7 days with change of medium every two days. These hES-T3 derived pancreatic islet-like cell clusters were designated as T3pi.
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	Trizol extraction of total RNA was performed according to the manufacturer's instructions. Total RNAs extracted using TRIZOL reagent were quantified spectrophotometrically. The cDNA synthesis was carried out by using SUPERSCRIPTTM one-step RT-PCR kit (Invitrogen). For the reverse transcription step, the whole 5 ul of the resuspended RNAs were incubated for 60 min at 42oC, then 15 min at 72oC in 50 ul of reaction mixture containing 25 ul of 2x Reaction Mix (Invitrogen) and 1 ul of RT/PLANTINUM Taq Mix (Invitrogen). 28.5 ul of the cDNAs present in the 50 ul RT reaction mixture purified by Microarray Target Purification Kit (Roche Applied Science, Indianapolis, ID, USA, http://www.roche-applies-science.com) were used as templates to amplify cDNA by Microarray Target Amplification Kit. The amplified cDNAs were purified by Microarray Target Purification Kit, and complementary RNAs (cRNAs) were synthesized from 200 ng cDNA with Microarray RNA Target Synthesis Kit (Roche Applied Science).
Sample_label_ch1	biotin
Sample_label_protocol_ch1	Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5 micrograms purified cRNA (Expression Analysis Technical Manual, 2004, Affymetrix, http://www.affymetrix.com).
Sample_hyb_protocol	Following fragmentation, 10 micrograms of cRNA were hybridized for 16 hr at 45oC on Affymetrix Human Genome U133 plus 2.0 GeneChip. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
Sample_scan_protocol	GeneChips from the hybridization experiments were read by the Affymetrix GeneChip scanner 3000.
Sample_data_processing	The data were processed by GeneSpring GX software version 7.3.1 GC-RMA preprocessor to use on the CEL files
Sample_platform_id	GPL570
Sample_contact_name	Sung-Liang,,Yu
Sample_contact_email	slyu@ntu.edu.tw
Sample_contact_phone	886-2-23958341
Sample_contact_fax	886-2-23958341
Sample_contact_laboratory	Microarray Core Facility
Sample_contact_department	Clinical Laboratory Sciences and Medical Biotechnology
Sample_contact_institute	National Taiwan University
Sample_contact_address	Jen Ai Road Section1
Sample_contact_city	Taipei
Sample_contact_zip/postal_code	100
Sample_contact_country	Taiwan
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM362nnn/GSM362249/suppl/GSM362249.CEL.gz
Sample_series_id	GSE14503
Sample_data_row_count	54675

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