RESEARCH ARTICLE

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

  • Lei XIAO , 1,2 ,
  • Lixiazi HE 2 ,
  • Saul J. SHARKIS , 1
Expand
  • 1. Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA
  • 2. Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China

Received date: 10 Sep 2010

Accepted date: 08 Oct 2010

Published date: 01 Oct 2010

Copyright

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg

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.

Cite this article

Lei XIAO , Lixiazi HE , Saul J. SHARKIS . Screening for self-renewal factors by a combination of mRNA and CGH microarray in human embryonic stem cells[J]. Frontiers in Biology, 2010 , 5(5) : 431 -438 . DOI: 10.1007/s11515-010-0900-1

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.
1
Ambrosini G, Adida C, Altieri D C (1997). A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma. Nat Med, 3(8): 917–921

DOI

2
Amit M, Shariki, C. Margulets V, Itskovitz-Eldor J (2004). Feeder layer- and serum-free culture of human embryonic stem cells. Biol Reprod, 70(3): 837–845

DOI

3
Buehr M, Meek S, Blair K, Yang J, Ure J, Silva J, McLay R, Hall J, Ying Q L, Smith A (2008). Capture of authentic embryonic stem cells from rat blastocysts. Cell, 135(7): 1287–1298

4
Chang Y F, Cheng C M, Chang L K, Jong Y J, Yuo C Y (2006). The F-box protein Fbxo7 interacts with human inhibitor of apoptosis protein cIAP1 and promotes cIAP1 ubiquitination. Biochem Biophys Res Commun, 342(4): 1022–1026

DOI

5
Chen G, Liu D, Tadokoro M, Hirochika R, Ohgushi H, Tanaka J, Tateishi T (2004). Chondrogenic differentiation of human mesenchymal stem cells cultured in a cobweb-like biodegradable scaffold. Biochem Biophys Res Commun, 322(1): 50–55

DOI

6
Coumoul X, Li W, Wang R H, Deng C (2004). Inducible suppression of Fgfr2 and Survivin in ES cells using a combination of the RNA interference (RNAi) and the Cre-LoxP system. Nucleic Acids Res, 32(10): e85

DOI

7
Draper J S, Smith K, Gokhale P, Moore H D, Maltby E, Johnson J, Meisner L, Zwaka T P, Thomson J A, Andrews P W (2004). Recurrent gain of chromosomes 17q and 12 in cultured human embryonic stem cells. Nat Biotechnol, 22(1): 53–54

DOI

8
Dravid G, Ye Z, Hammond H, Chen G, Pyle A, Donovan P, Yu X, Cheng L (2005). Defining the role of Wnt/beta-catenin signaling in the survival, proliferation, and self-renewal of human embryonic stem cells. Stem Cells, 23(10): 1489–1501

DOI

9
Guo Y, Mantel C, Hromas R A, Broxmeyer H E (2008). Oct-4 is critical for survival/antiapoptosis of murine embryonic stem cells subjected to stress: effects associated with Stat3/survivin. Stem Cells, 26(1): 30–34

DOI

10
He Z, Feng L, Zhang X, Geng Y, Parodi D A, Suarez-Quian C, Dym M (2005). Expression of Col1a1, Col1a2 and procollagen I in germ cells of immature and adult mouse testis. Reproduction, 130(3): 333–341

DOI

11
Ikeda M, Hikita T, Taya S, Uraguchi-Asaki J, Toyo-oka K, Wynshaw-Boris A, Ujike H, Inada T, Takao K, Miyakawa T, Ozaki N, Kaibuchi K, Iwata N (2008). Identification of YWHAE, a gene encoding 14-3-3epsilon, as a possible susceptibility gene for schizophrenia. Hum Mol Genet, 17(20): 3212–3222

DOI

12
Jacobs J J, Keblusek P, Robanus-Maandag E, Kristel P, Lingbeek M, Nederlof P M, van Welsem T, van de Vijver M J, Koh E Y, Daley G Q, van Lohuizen M (2000). Senescence bypass screen identifies TBX2, which represses Cdkn2a (p19(ARF)) and is amplified in a subset of human breast cancers. Nat Genet, 26(3): 291–299

DOI

13
Lee S K, Park S O, Joe C O, Kim Y S (2007). Interaction of HCV core protein with 14-3-3epsilon protein releases Bax to activate apoptosis. Biochem Biophys Res Commun, 352(3): 756–762

DOI

14
Li P, Tong C, Mehrian-Shai R, Jia L, Wu N, Yan Y, Maxson R E, Schulze E N, Song H, Hsieh C L, Pera M F, Ying Q L (2008). Germline competent embryonic stem cells derived from rat blastocysts. Cell, 135(7): 1299–1310

15
Maitra A, Arking D E, Shivapurkar N, Ikeda M, Stastny V, Kassauei K, Sui G, Cutler D J, Liu Y, Brimble S N, Noaksson K, Hyllner J, Schulz T C, Zeng X, Freed W J, Crook J, Abraham S, Colman A, Sartipy P, Matsui S, Carpenter M, Gazdar A F, Rao M, Chakravarti A (2005). Genomic alterations in cultured human embryonic stem cells. Nat Genet, 37(10): 1099–1103

DOI

16
Mitalipova M M, Rao R R, Hoyer D M, Johnson J A, Meisner L F, Jones K L, Dalton S, Stice S L (2005). Preserving the genetic integrity of human embryonic stem cells. Nat Biotechnol, 23(1): 19–20

DOI

17
Mitelman F (1995). An International System for Human Cytogenetic Nomenclature in ISCN. Basel, Karger AG

18
O’Connor D S, Grossman D, Plescia J, Li F, Zhang H, Villa A, Tognin S, Marchisio P C, Altieri D C (2000). Regulation of apoptosis at cell division by p34cdc2 phosphorylation of survivin. Proc Natl Acad Sci U S A, 97(24): 13103–13107

DOI

19
Pease S, Braghetta P, Gearing D., Grail D, Williams R L (1990). Isolation of embryonic stem (ES) cells in media supplemented with recombinant leukemia inhibitory factor (LIF). Dev Biol, 141(2): 344–352

DOI

20
Prakash O, Yunis J J (1984). High resolution chromosomes of the t(9;22) positive leukemias. Cancer Genet Cytogenet, 11(4): 361–367

DOI

21
Sato N, Meijer L, Skaltsounis L, Greengard P, Brivanlou A H (2004). Maintenance of pluripotency in human and mouse embryonic stem cells through activation of Wnt signaling by a pharmacological GSK-3-specific inhibitor. Nat Med, 10(1): 55–63

DOI

22
Schröck E, du Manoir S, Veldman T, Schoell B, Wienberg J, Ferguson-Smith M A, Ning Y, Ledbetter D H, Bar-Am I, Soenksen D, Garini Y, Ried T (1996). Multicolor spectral karyotyping of human chromosomes. Science, 273(5274): 494–497

DOI

23
Shtivelman E, Lifshitz B, Gale R P, Canaani E (1985). Fused transcript of abl and bcr genes in chronic myelogenous leukaemia. Nature, 315(6020): 550–554

DOI

24
Silva C, Wood J R, Salvador L, Zhang Z, Kostetskii I, Williams C J, Strauss J F 3rd (2009). Expression profile of male germ cell-associated genes in mouse embryonic stem cell cultures treated with all-trans retinoic acid and testosterone. Mol Reprod Dev, 76(1): 11–21

DOI

25
Thomson J A, Itskovitz-Eldor J, Shapiro S S, Waknitz M A, Swiergiel J J, Marshall V S, Jones J M (1998). Embryonic stem cell lines derived from human blastocysts. Science, 282(5391): 1145–1147

DOI

26
Vance K W, Carreira S, Brosch G, Goding C R (2005). Tbx2 is overexpressed and plays an important role in maintaining proliferation and suppression of senescence in melanomas. Cancer Res, 65(6): 2260–2268

DOI

27
Wang Y, Hai T, Liu Z, Zhou S, Lv Z, Ding C, Liu L, Niu Y, Zhao X, Tong M, Wang L, Jouneau A, Zhang X, Ji W, Zhou Q (2010). HSPC117 deficiency in cloned embryos causes placental abnormality and fetal death. Biochem Biophys Res Commun, 397(3): 407–412

DOI

28
Xiao L, Yuan X, Sharkis S J (2006). Activin A maintains self-renewal and regulates fibroblast growth factor, Wnt, and bone morphogenic protein pathways in human embryonic stem cells. Stem Cells, 24(6): 1476–1486

DOI

29
Xu C, Inokuma M S, Denham J, Golds K, Kundu P, Gold J D, Carpenter M K (2001). Feeder-free growth of undifferentiated human embryonic stem cells. Nat Biotechnol, 19(10): 971–974

DOI

30
Yamada M, Ohkawara B, Ichimura N, Hyodo-Miura J, Urushiyama S, Shirakabe K., Shibuya H (2003). Negative regulation of Wnt signalling by HMG2L1, a novel NLK-binding protein. Genes Cells, 8(8): 677–684

DOI

31
Ying Q L, Nichols J, Chambers I, Smith A (2003). BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell, 115(3): 281–292

32
Zeng X, Chen J, Liu Y, Luo Y, Schulz T C, Robins A J, Rao M S, Freed W J (2004). BG01V: a variant human embryonic stem cell line which exhibits rapid growth after passaging and reliable dopaminergic differentiation. Restor Neurol Neurosci, 22(6): 421–428

33
Zhou J, Hu G, Wang X (2010). Repression of smooth muscle differentiation by a novel high mobility group box-containing protein, HMG2L1. J Biol Chem, 285(30): 23177–23185

DOI

Outlines

/