Overexpression of bone morphogenetic protein 4 in STO fibroblast feeder cells represses the proliferation of mouse embryonic stem cells in vitro

다국어 초록 (Multilingual Abstract)

Embryonic stem cells (ESCs) can be propagated in vitro on feeder layers of mouse STO fibroblast cells. The STO cells secrete several cytokines that are essential for ESCs to maintain their undifferentiated state. In this study, we found significant growth inhibition of mouse ESCs (mESCs) cultured on STO cells infected with adenovirus containing a dominant-negative mutant form of IκB (rAd-dnIκB). This blockage of the NF-κB signal pathway in STO cells led to a significant decrease in [3H]thymidine incorporation and colony formation of mESCs. Expression profile of cytokines secreted from the STO cells revealed an increase in the bone morphogenetic protein4 (BMP4) transcript level in the STO cells infected with adenoviral vector encoding dominant negative IκB (rAd-dnIκB). These results suggested that the NF-κB signaling pathway represses expression of BMP4 in STO feeder cells. Conditioned medium from the rAd-dnIκB-infected STO cells also significantly reduced the colony size of mESCs. Addition of BMP4 prevented colony formation of mESCs cultured in the conditioned medium. Our finding suggested that an excess of BMP4 in the conditioned medium also inhibits proliferation of mESCs.

참고문헌 (Reference)

1 Hao J, "WNT/beta-catenin pathway up-regulates Stat3 and converges on LIF to prevent differentiation of mouse embryonic stem cells" 290 : 81-91, 2006

2 Pearson S, "The stepwise specification of embryonic stem cells to hematopoietic fate is driven by sequential exposure to Bmp4, activin A, bFGF and VEGF" 135 : 1525-1535, 2008

3 Zhu NL, "TNF-alpha represses transcription of human Bone Morphogenetic Protein-4 in lung epithelial cells" 393 : 70-80, 2007

4 Bhardwaj G, "Sonic hedgehog induces the proliferation of primitive human hematopoietic cells via BMP regulation" 2 : 172-178, 2001

5 Niwa H, "Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or selfrenewal of ES cells" 24 : 372-376, 2000

6 Bhatia M, "Quantitative analysis reveals expansion of human hematopoietic repopulating cells after short-term ex vivo culture" 186 : 619-624, 1997

7 Fainsod A, "On the function of BMP-4 in patterning the marginal zone of the Xenopus embryo" 13 : 5015-5025, 1994

8 Reese DE, "Negative regulation of midline vascular development by the notochord" 6 : 699-708, 2004

9 Muraoka RS, "Mesenchymal expression of nuclear factor-kappaB inhibits epithelial growth and branching in the embryonic chick lung" 225 : 322-338, 2000

10 Vicente Lopez MA, "Low doses of bone morphogenetic protein 4 increase the survival of human adipose-derived stem cells maintaining their stemness and multipotency" 20 : 1011-1019, 2011

11 Jones CM, "Involvement of Bone Morphogenetic Protein-4 (BMP-4) and Vgr-1 in morphogenesis and neurogenesis in the mouse" 111 : 531-542, 1991

12 Ji-Yeon Ahn, "Intramarrow injection of β-catenin-activated, but not naïve mesenchymal stromal cells stimulates self-renewal of hematopoietic stem cells in bone marrow" 생화학분자생물학회 42 (42): 122-131, 2010

13 Smith AG, "Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides" 336 : 688-690, 1988

14 Vainio S, "Identification of BMP-4 as a signal mediating secondary induction between epithelial and mesenchymal tissues during early tooth development" 75 : 45-58, 1993

15 Hollnagel A, "Id genes are direct targets of bone morphogenetic protein induction in embryonic stem cells" 274 : 19838-19845, 1999

16 Murdoch B, "Human embryonic-derived hematopoietic repopulating cells require distinct factors to sustain in vivo repopulating function" 30 : 598-605, 2002

17 Smith AG, "Embryo-derived stem cells: of mice and men" 17 : 435-462, 2001

18 Schulz TC, "Differentiation of human embryonic stem cells to dopaminergic neurons in serum-free suspension culture" 22 : 1218-1238, 2004

19 Mazaheri Z, "Different doses of bone morphogenetic protein 4 promote the expression of early germ cell-specific gene in bone marrow mesenchymal stem cells. in vitro" 47 : 521-525, 2011

20 Brons IG, "Derivation of pluripotent epiblast stem cells from mammalian embryos" 448 : 191-195, 2007

21 Schenke-Layland K, "Collagen IV induces trophoectoderm differentiation of mouse embryonic stem cells" 25 : 1529-1538, 2007

22 Bhatia M, "Bone morphogenetic proteins regulate the developmental program of human hematopoietic stem cells" 189 : 1139-1148, 1999

23 Xu RH, "Basic FGF and suppression of BMP signaling sustain undifferentiated proliferation of human ES cells" 2 : 185-190, 2005

24 Qi X, "BMP4 supports self-renewal of embryonic stem cells by inhibiting mitogen-activated protein kinase pathways" 101 : 6027-6032, 2004

25 Xu RH, "BMP4 initiates human embryonic stem cell differentiation to trophoblast" 20 : 1261-1264, 2002

26 Ying QL, "BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3" 115 : 281-292, 2003

27 Livak KJ, "Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method" 25 : 402-408, 2001

28 Yong Bin Park, "Alterations of proliferative and differentiation potentials ofhuman embryonic stem cells during long-term culture" 생화학분자생물학회 40 (40): 98-108, 2008

29 De Robertis EM, "A common plan for dorsoventral patterning in Bilateria" 380 : 37-40, 1996