Differential genes in adipocytes induced from polycystic and non-polycystic ovary syndrome-derived human embryonic stem cells.

Differential genes in adipocytes induced from polycystic and non-polycystic ovary syndrome-derived human embryonic stem cells.

Systems biology in reproductive medicine (2014-02-13)

Fang Wang, Wei-Wei Liu, Xue-Mei Chen, Hui-Juan Kong, Jing Li, Ying-Pu Sun

ABSTRACT

We explored the molecular mechanisms of obesity and insulin resistance in patients with polycystic ovary syndrome (PCOS) using a human embryonic stem cell model (hESCs). Three PCOS-derived and one non-PCOS-derived hESC lines were induced into adipocytes, and then total RNA was extracted. The differentially expressed PCOS-derived and non-PCOS-derived adipocytes genes were identified using the Boao Biological human V 2.0 whole genome oligonucleotide microarray. Signals of interest were then validated by real-time PCR. A total of 153 differential genes were expressed of which 91 genes were up-regulated and 62 down-regulated. Nuclear receptor subfamily 0, group B, member 2 (NR0B2) was an up-regulated gene, and the GeneChip CapitalBio® Molecule Annotation System V4.0 indicated that it was associated with obesity and diabetes (Ratio ≥ 2.0X). Multiple genes are involved in PCOS. Nuclear receptor subfamily 0, group B, member 2 may play a role in obesity and insulin resistance in patients with PCOS.

MATERIALS

Product Number

Brand

Product Description

209651

Sigma-Aldrich

Selenium, powder, −100 mesh, ≥99.5% trace metals basis

229865

Sigma-Aldrich

Selenium, powder, −100 mesh, 99.99% trace metals basis

209643

Sigma-Aldrich

Selenium, pellets, <5 mm, ≥99.99% trace metals basis

204307

Sigma-Aldrich

Selenium, pellets, <5 mm particle size, ≥99.999% trace metals basis

I7018

Sigma-Aldrich

3-Isobutyl-1-methylxanthine, BioUltra, ≥99%

I5879

Sigma-Aldrich

3-Isobutyl-1-methylxanthine, ≥99% (HPLC), powder

R2408

Sigma-Aldrich

Rosiglitazone

H7523

Sigma-Aldrich

HEPES, BioXtra, pH 5.0-6.5 (1 M in H₂O), ≥99.5% (titration)

GF66395212

Selenium, foil, 25x25mm, thickness 3mm, 99.95%

H3375

Sigma-Aldrich

HEPES, ≥99.5% (titration)

54457

Sigma-Aldrich

HEPES, BioUltra, Molecular Biology, ≥99.5% (T)

H4034

Sigma-Aldrich

HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture

H6147

Sigma-Aldrich

HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)

PHR1428

Supelco

HEPES, Pharmaceutical Secondary Standard; Certified Reference Material

RDD002

Sigma-Aldrich

HEPES, anhydrous, free-flowing, Redi-Dri^™, ≥99.5%

V900477

Sigma-Aldrich

HEPES, Vetec^™, reagent grade, 99.5%

D4902

Sigma-Aldrich

Dexamethasone, powder, BioReagent, suitable for cell culture, ≥97%

E8008

Sigma-Aldrich

Ethylenediaminetetraacetic acid solution, 0.02% in DPBS (0.5 mM), sterile-filtered, BioReagent, suitable for cell culture

74875

Sigma-Aldrich

8-Octanoyloxypyrene-1,3,6-trisulfonic acid trisodium salt, suitable for fluorescence, ≥90% (HPCE)

276855

Sigma-Aldrich

Dimethyl sulfoxide, anhydrous, ≥99.9%

83264

Sigma-Aldrich

HEPES buffer solution, 1 M in H₂O

431788

Sigma-Aldrich

Ethylenediaminetetraacetic acid, 99.995% trace metals basis

94563

Supelco

Dimethyl sulfoxide, analytical standard

41640

Sigma-Aldrich

Dimethyl sulfoxide, puriss. p.a., ACS reagent, ≥99.9% (GC)

472301

Sigma-Aldrich

Dimethyl sulfoxide, ACS reagent, ≥99.9%

01934

Supelco

Dimethyl sulfoxide, for inorganic trace analysis, ≥99.99995% (metals basis)

1211006

USP

Dimethyl sulfoxide, United States Pharmacopeia (USP) Reference Standard

41639

Sigma-Aldrich

Dimethyl sulfoxide, BioUltra, Molecular Biology, ≥99.5% (GC)

D4540

Sigma-Aldrich

Dimethyl sulfoxide, ≥99.5% (GC), suitable for plant cell culture

D2438

Sigma-Aldrich

Dimethyl sulfoxide, sterile-filtered, BioPerformance Certified, meets EP, USP testing specifications, suitable for hybridoma