Screening for self-renewal factors by a combination of mRNA and CGH microarray in human embryonic stem cells

Abstract

Human embryonic stem cells (hESCs) undergo self-renewal while maintaining pluripotency. However, the molecular mechanism that demonstrates how these cells maintain their undifferentiated state and how they self-renew is poorly understood. Here, we characterized an aneuploidy H1 hESC subline (named H1T) using karyotyping and comparative genomic hybridization (CGH) microarray. Because the H1T hESC line displays a self-renewal advantage while maintaining an undifferentiated state, we speculated that the expression patterns of specific genes which are related to pluripotency or differentiation were altered; therefore, we attempted to screen for molecules that are propitious for maintenance of stemness by performing a combination of mRNA and CGH microarray analysis which compared the aneuploidy H1T hESC subline versus the euploid H1 hESC line. It is discovered that some genes are up-regulated in H1T hESC subline such as TBX2 and Wnt3, while some are downregulated, for example, Fbxo7 and HMG2L1. Our findings should fascilitate the study of the complex signaling network which maintains hESC pluripotency and function.

Keywords

self-renewal / mRNA microarray / comparative genomic hybridization (CGH) microarray

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Acknowledgements

We thank A. HAWKINS and L. MORSBERGER from Cytogenetics Core Facility for karyotyping; Dr. W. YU, H. XU, and L. SUO from the SKCCC Microarray Core Facility for microarray analysis. This work was supported by a grant from the NIH PO1 Ca70970. This research was partially supported by grants from the National Natural Science Foundation of China (Grant No. 31025016). The authors declare that they have no competing financial interests.

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2014 Higher Education Press and Springer-Verlag Berlin Heidelberg