TOP TEN perturbations for NM_000029 (Homo sapiens)

Organism: Homo sapiens
Gene: NM_000029
Selected probe(set): 202834_at
Platform: Affymetrix Human Genome U133 Plus 2.0 Array

Expression of NM_000029 (202834_at) across 5392 perturbations tested by GENEVESTIGATOR:

HCC study 20 (CDX; Hep-G2; orthotopic) / HCC study 20 (PDX; orthotopic)

Relative Expression (log2-ratio):-8.437394
Number of Samples:5 / 11
Experimental HCC study 20 (CDX; Hep-G2; orthotopic)
Tumor tissue biopsy samples from Hep-G2 cell line-derived xenograft (CDX) orthotopically generated in SCID mice by injecting hepatocellular carcinoma cells directly into liver parenchyma. In order to establish orthotopic model, the left lobe of liver was exposed through midline abdominal incision and cells were injected directly into liver parenchyma of mice. The mice were euthanized after two weeks.
Control HCC study 20 (PDX; orthotopic)
Tumor tissue biopsy samples from patient-derived xenograft (PDX) sample orthotopically generated in SCID mice by injecting hepatocellular carcinoma cells directly into liver parenchyma. Donor tumor tissue was obtained intraoperatively during liver resection from three patients. All three patients had hepatocellular carcinoma confirmed by histology. In order to establish orthotopic model, the left lobe of liver was exposed through midline abdominal incision and cells were injected directly into liver parenchyma of mice. The mice were euthanized after two weeks.

HCC study 20 (CDX; Hep-G2; ectopic) / HCC study 20 (PDX; ectopic)

Relative Expression (log2-ratio):-8.138589
Number of Samples:3 / 11
Experimental HCC study 20 (CDX; Hep-G2; ectopic)
Tumor tissue biopsy samples from Hep-G2 cell line-derived xenograft (CDX) ectopically generated in SCID mice by injecting hepatocellular carcinoma cells subcutaneously. In order to establish ectopic models, cells were injected subcutaneously into the flank regions. The mice were euthanized after two weeks.
Control HCC study 20 (PDX; ectopic)
Tumor tissue biopsy samples from patient-derived xenograft (PDX) sample ectopically generated in SCID mice by injecting hepatocellular carcinoma cells subcutaneously. Donor tumor tissue was obtained intraoperatively during liver resection from three patients. All three patients had hepatocellular carcinoma confirmed by histology. In order to establish ectopic models, cells were injected subcutaneously into the flank regions. The mice were euthanized after two weeks.

hepatocyte-like cell differentiation study 1 (15d) / hepatocyte-like cell differentiation study 1 (5d)

Relative Expression (log2-ratio):5.3259287
Number of Samples:3 / 3
Experimental hepatocyte-like cell differentiation study 1 (15d)
Hepatocyte-like cells differentiated from human embryonic stem cells (ES, WA09), 15 days after the onset of differentiation. This stage is approximately corresponding to immature hepatocytes. Human H9 (WA09) ES cells were routinely cultured under low oxygen conditions (4% O2/5% CO2) in human ES cell media (DMEM/F12 medium supplemented with 20% knockout serum replacement, non essential amino acids, glutamine, penicillin/streptomycin and bFGF (4ng/ml) on Matrigel coated plates using MEF-conditioned human ES cell media. Differentiation was initiated by culture for 5 days with 100ng/ml Activin A in RPMI/B27 medium under ambient oxygen/5%CO2 followed by 5 days with 20ng/ml BMP4 /10ng/ml FGF-2 in RPMI/B27 under 4%O2/5%CO2, then 5 days with 20ng/ml HGF in RPMI/B27 supplement under 4%O2/5%CO2.
Control hepatocyte-like cell differentiation study 1 (5d)
Hepatocyte-like cell differentiated from human pluripotent, embryonic stem cell line (ES, WA09). ES cells were grown in human ES cell media DMEM/F12 and differentiated to definitive endoderm for 5 days.

hepatocyte-like cell differentiation study 1 (15d) / hepatocyte-like cell differentiation study 1 (10d)

Relative Expression (log2-ratio):5.210246
Number of Samples:3 / 3
Experimental hepatocyte-like cell differentiation study 1 (15d)
Hepatocyte-like cells differentiated from human embryonic stem cells (ES, WA09), 15 days after the onset of differentiation. This stage is approximately corresponding to immature hepatocytes. Human H9 (WA09) ES cells were routinely cultured under low oxygen conditions (4% O2/5% CO2) in human ES cell media (DMEM/F12 medium supplemented with 20% knockout serum replacement, non essential amino acids, glutamine, penicillin/streptomycin and bFGF (4ng/ml) on Matrigel coated plates using MEF-conditioned human ES cell media. Differentiation was initiated by culture for 5 days with 100ng/ml Activin A in RPMI/B27 medium under ambient oxygen/5%CO2 followed by 5 days with 20ng/ml BMP4 /10ng/ml FGF-2 in RPMI/B27 under 4%O2/5%CO2, then 5 days with 20ng/ml HGF in RPMI/B27 supplement under 4%O2/5%CO2.
Control hepatocyte-like cell differentiation study 1 (10d)
Hepatocyte-like cell differentiated from human pluripotent, embryonic stem cell line (ES, WA09). ES cells were grown in human ES cell media DMEM/F12 and differentiated to hepatic specification stage for 10 days.

hepatocyte-like cell differentiation study 1 (20d; HNF4 depl) / hepatocyte-like cell differentiation study 1 (15d)

Relative Expression (log2-ratio):-5.161315
Number of Samples:3 / 3
Experimental hepatocyte-like cell differentiation study 1 (20d; HNF4 depl)
Hepatocyte-like cell differentiated from human pluripotent, embryonic stem cell line (ES, WA09) and transfected with shRNA targeting HNF4A. ES cells were grown in human ES cell media DMEM/F12 and differentiated to mature hepatocytes for 20 days. Moreover hESc was stable transfected with lentivirus expressing an shRNA that efficiently depletes HNF4A.
Control hepatocyte-like cell differentiation study 1 (15d)
Hepatocyte-like cells differentiated from human embryonic stem cells (ES, WA09), 15 days after the onset of differentiation. This stage is approximately corresponding to immature hepatocytes. Human H9 (WA09) ES cells were routinely cultured under low oxygen conditions (4% O2/5% CO2) in human ES cell media (DMEM/F12 medium supplemented with 20% knockout serum replacement, non essential amino acids, glutamine, penicillin/streptomycin and bFGF (4ng/ml) on Matrigel coated plates using MEF-conditioned human ES cell media. Differentiation was initiated by culture for 5 days with 100ng/ml Activin A in RPMI/B27 medium under ambient oxygen/5%CO2 followed by 5 days with 20ng/ml BMP4 /10ng/ml FGF-2 in RPMI/B27 under 4%O2/5%CO2, then 5 days with 20ng/ml HGF in RPMI/B27 supplement under 4%O2/5%CO2.

pancreatic islet study 3 (re-differentiated; PPRF; 2d) / pancreatic islet study 3 (expanded; PPRF)

Relative Expression (log2-ratio):5.0837383
Number of Samples:2 / 2
Experimental pancreatic islet study 3 (re-differentiated; PPRF; 2d)
Human pancreatic islets were isolated from a cadaveric donor. The population was 70% pure in mature islets, which was determined by dithizone staining. Islets cells were expanded for 6 weeks and re-differentiated for 2 days according to Pharmaceutical Production Research Facility (PPRF) Protocol. Re-differentiation phase: For induction of expanded cells to differentiate into islet-like cell clusters, the cells were seeded into 60 mm ultra-low attachment dishes in serum-free CMRL-1066 supplemented with 4 mM L-glutamine, 1% BSA, insulin (10 g/ml), transferrin (5.5 g/ml), sodium selenite (6.7ng/ml), and 1% antibiotic antimycotic solution at a density of 6.0 x 10EXP6 cells per dish. The induction medium was further added with islet neogenesis-associated protein (INGAP) peptide at a concentration of 1.0ug/ml.
Control pancreatic islet study 3 (expanded; PPRF)
Human pancreatic islets were isolated from a cadaveric donor. The population was 70% pure in mature islets, which was determined by dithizone staining. Islets cells were expanded according to Pharmaceutical Production Research Facility Protocol (PPRF) for 6 weeks. Expansion phase: Approximately 2,500 islet equivalents were seeded onto 75-cm2 fibronectin-coated tissue culture-treated flasks in a proprietary serum-free medium (PPRF medium). The serum-free PPRF medium was supplemented with 30% serum-free mesenchymal-conditioned medium. When the cell migration resulted in the formation of large-size colonies, the cells were harvested by trypsinization and replated into new tissue culture flasks. Once the cells reached near confluence (80?90%) in monolayer, they were trypsinized, counted, and subcultured at a density of 5,000 cells per cm2. Thereafter, cells were serially passaged using the same protocol.

glioma study 16 (BS-149) / normal astrocyte sample

Relative Expression (log2-ratio):4.718837
Number of Samples:2 / 3
Experimental glioma study 16 (BS-149)
Human glioma cell line BS149 was cultured in DMEM supplemented with 10% FCS (fetal calf serum) and antibiotics at 37?C and 5% CO2.
Control normal astrocyte sample
Normal brain astrocytes. Clonetics normal human astrocytes (NHA) from Cambrex (primary-derived cultures) and cultured according to the manufacturer's recommendations.

pancreatic islet study 3 (re-differentiated; PPRF; 4d) / pancreatic islet study 3 (expanded; PPRF)

Relative Expression (log2-ratio):4.4252605
Number of Samples:2 / 2
Experimental pancreatic islet study 3 (re-differentiated; PPRF; 4d)
Human pancreatic islets were isolated from a cadaveric donor. The population was 70% pure in mature islets, which was determined by dithizone staining. Islets cells were expanded for 6 weeks and re-differentiated for 4 days according to Pharmaceutical Production Research Facility (PPRF) Protocol. Re-differentiation phase: For induction of expanded cells to differentiate into islet-like cell clusters, the cells were seeded into 60 mm ultra-low attachment dishes in serum-free CMRL-1066 supplemented with 4 mM L-glutamine, 1% BSA, insulin (10 g/ml), transferrin (5.5 g/ml), sodium selenite (6.7ng/ml), and 1% antibiotic antimycotic solution at a density of 6.0 x 10EXP6 cells per dish. The induction medium was further added with islet neogenesis-associated protein (INGAP) peptide at a concentration of 1.0ug/ml.
Control pancreatic islet study 3 (expanded; PPRF)
Human pancreatic islets were isolated from a cadaveric donor. The population was 70% pure in mature islets, which was determined by dithizone staining. Islets cells were expanded according to Pharmaceutical Production Research Facility Protocol (PPRF) for 6 weeks. Expansion phase: Approximately 2,500 islet equivalents were seeded onto 75-cm2 fibronectin-coated tissue culture-treated flasks in a proprietary serum-free medium (PPRF medium). The serum-free PPRF medium was supplemented with 30% serum-free mesenchymal-conditioned medium. When the cell migration resulted in the formation of large-size colonies, the cells were harvested by trypsinization and replated into new tissue culture flasks. Once the cells reached near confluence (80?90%) in monolayer, they were trypsinized, counted, and subcultured at a density of 5,000 cells per cm2. Thereafter, cells were serially passaged using the same protocol.

pancreatic islet study 3 (re-differentiated; PPRF; 2d) / pancreatic islet study 3 (re-differentiated; NIH)

Relative Expression (log2-ratio):4.3613796
Number of Samples:2 / 4
Experimental pancreatic islet study 3 (re-differentiated; PPRF; 2d)
Human pancreatic islets were isolated from a cadaveric donor. The population was 70% pure in mature islets, which was determined by dithizone staining. Islets cells were expanded for 6 weeks and re-differentiated for 2 days according to Pharmaceutical Production Research Facility (PPRF) Protocol. Re-differentiation phase: For induction of expanded cells to differentiate into islet-like cell clusters, the cells were seeded into 60 mm ultra-low attachment dishes in serum-free CMRL-1066 supplemented with 4 mM L-glutamine, 1% BSA, insulin (10 g/ml), transferrin (5.5 g/ml), sodium selenite (6.7ng/ml), and 1% antibiotic antimycotic solution at a density of 6.0 x 10EXP6 cells per dish. The induction medium was further added with islet neogenesis-associated protein (INGAP) peptide at a concentration of 1.0ug/ml.
Control pancreatic islet study 3 (re-differentiated; NIH)
Pancreatic islet cells were expanded for 10 weeks and re-differentiated for 1 week according to National Institutes of Health (NIH) protocol. Re-differentiation phase: Expanded cells were cultured for 1 week in serum-free CMRL-1066 medium supplemented with insulin (10 g/ml), transferrin (5.5 g/ml), sodium selenite (6.7ng/ml).

pancreatic islet study 3 (expanded; NIH) / normal pancreatic islet sample

Relative Expression (log2-ratio):-4.310749
Number of Samples:3 / 7
Experimental pancreatic islet study 3 (expanded; NIH)
Pancreatic islet cells were expanded according to National Institutes of Health (NIH) protocol for 10 weeks. Expansion phase: 2,000 islet equivalents enriched by retention on a 40-?m filter were seeded onto tissue culture?treated dishes in CMRL-1066 medium containing 2 mmol/l l-glutamine and 10% fetal bovine serum.
Control normal pancreatic islet sample
Pancreatic islets were obtained from seven donors aged between 37 and 70 years and body mass index between 22 and 27. Functional islets were cultured in CMRL 1066 supplemented with 10% FCS, 1% glutamine, 5.6 mM glucose, 1 mM HEPES, 110 U/ml penicillin, and 110 g/ml streptomycin for 7 days or immediately processed after isolation.