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Lipid rafts play an important role for maintenance of embryonic stem cell self-renewal.

ABSTRACT: Lipid rafts are cholesterol-rich microdomains of cell membranes that have a variety of roles in cellular processes including receptor-mediated signal transduction. Lipid rafts also occur in embryonic stem (ES) cells, but their role in ES cells is largely unknown. Therefore, we investigated the role of lipid rafts in the maintenance of ES cell self-renewal. In the present study, we observed that the presence of lipid rafts/caveolae. The results from sucrose gradient fractionation showed that the expression of glycoprotein 130 (gp130) and leukemia inhibitory factor receptor beta (LIFRbeta) was decreased by treatment with methyl-beta-cyclodextrin (Mbeta-CD) but, interestingly, was not affected by caveolin-1 small interfering RNA (siRNA). In addition, LIF increased phosphorylation of signal transducer and activator of transcription 3 (STAT3) and Akt, and the expression level of c-Myc, which were attenuated by the pretreatment with Mbeta-CD. However, caveolin-1 siRNA did not influence LIF-induced phosphorylation of STAT3 and Akt, and expression of c-Myc. Treatment with Mbeta-CD and caveolin-1 siRNA decreased expression levels of Oct4 protein and Oct4, Sox2, FoxD3, and Rex1 mRNAs in normal culture conditions. Additionally, Mbeta-CD and caveolin-1 siRNA decreased the expression levels of cyclin D1 and cyclin E, and the proliferation index [(S + G2/M)/(G0/G1 + S + G2/M)] of ES cells. We conclude that lipid raft/caveolae structures play important roles in the self-renewal of ES cells.

SUBMITTER: Lee MY

PROVIDER: S-EPMC2903794 | biostudies-literature | 2010 Aug

REPOSITORIES: biostudies-literature

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Lipid rafts play an important role for maintenance of embryonic stem cell self-renewal.

Lee Min Young MY Ryu Jung Min JM Lee Sang Hun SH Park Jae Hong JH Han Ho Jae HJ

Journal of lipid research 20100128 8

Lipid rafts are cholesterol-rich microdomains of cell membranes that have a variety of roles in cellular processes including receptor-mediated signal transduction. Lipid rafts also occur in embryonic stem (ES) cells, but their role in ES cells is largely unknown. Therefore, we investigated the role of lipid rafts in the maintenance of ES cell self-renewal. In the present study, we observed that the presence of lipid rafts/caveolae. The results from sucrose gradient fractionation showed that the ...[more]

PMID: 20110442

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Project description:Mouse embryonic stem cells (mESCs) are derived from the inner cell mass of a developing blastocyst and can be cultured indefinitely in-vitro. Their distinct features are their ability to self-renew and to differentiate to all adult cell types. Genes that maintain mESCs self-renewal and pluripotency identity are of interest to stem cell biologists. Although significant steps have been made toward the identification and characterization of such genes, the list is still incomplete and controversial. For example, the overlap among candidate self-renewal and pluripotency genes across different RNAi screens is surprisingly small. Meanwhile, machine learning approaches have been used to analyze multi-dimensional experimental data and integrate results from many studies, yet they have not been applied to specifically tackle the task of predicting and classifying self-renewal and pluripotency gene membership.For this study we developed a classifier, a supervised machine learning framework for predicting self-renewal and pluripotency mESCs stemness membership genes (MSMG) using support vector machines (SVM). The data used to train the classifier was derived from mESCs-related studies using mRNA microarrays, measuring gene expression in various stages of early differentiation, as well as ChIP-seq studies applied to mESCs profiling genome-wide binding of key transcription factors, such as Nanog, Oct4, and Sox2, to the regulatory regions of other genes. Comparison to other classification methods using the leave-one-out cross-validation method was employed to evaluate the accuracy and generality of the classification. Finally, two sets of candidate genes from genome-wide RNA interference screens are used to test the generality and potential application of the classifier.Our results reveal that an SVM approach can be useful for prioritizing genes for functional validation experiments and complement the analyses of high-throughput profiling experimental data in stem cell research.

Dataset Information

Lipid rafts play an important role for maintenance of embryonic stem cell self-renewal.

Publications

Lipid rafts play an important role for maintenance of embryonic stem cell self-renewal.

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