Team:MIT mammalian Bone

From 2010.igem.org

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<b> Background </b>
<b> Background </b>
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The goal of this portion of the project was to synthetically differentiate stem cells into bone; this would act as the 'output' for our final system. Since stem cells can by tricky to genetically engineer, we chose to build our circuit in human endothelial kidney (HEK) cells. In the final system, HEK cells will secrete a diffusible morphogen to differentiate co-cultured stem cells. We used Bone Morphogenetic Protein 2 (BMP2) as our osteogenic signaling molecule; it is one of the central regulators of osteoblast differentiation in mammalian cells (1) and has been shown to induce transdifferentiation in multiple stem cell types (2) (3). Here, we work with myoblastic progenitor (C2C12) and mesenchymal (C3HT101/2) stem cells, two cell lines which have been shown capable of osteoblast differentiation.   
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The goal of this portion of the project was to synthetically differentiate stem cells into bone; this would act as the 'output' for our final system. Since stem cells can by tricky to genetically engineer, we chose to build our circuit in human endothelial kidney (HEK) cells. In the final system, HEK cells will secrete a diffusible morphogen to differentiate co-cultured stem cells. We used Bone Morphogenetic Protein 2 (BMP2) as our osteogenic signaling molecule; it is one of the central regulators of osteoblast differentiation in mammalian cells (1) and has been shown to induce transdifferentiation in multiple stem cell types (2, 3, 4). Here, we work with myoblastic progenitor (C2C12) and mesenchymal (C3HT101/2) stem cells, two cell lines which have been shown capable of osteoblast differentiation (5, 6).   
<a href="http://2010.igem.org/wiki/images/e/ed/Bone_ALP_Assay_7_6_2010-2.jpg" target="_blank"> <img width=600px src="http://2010.igem.org/wiki/images/e/ed/Bone_ALP_Assay_7_6_2010-2.jpg"> </a>
<a href="http://2010.igem.org/wiki/images/e/ed/Bone_ALP_Assay_7_6_2010-2.jpg" target="_blank"> <img width=600px src="http://2010.igem.org/wiki/images/e/ed/Bone_ALP_Assay_7_6_2010-2.jpg"> </a>
<b> References </b>
<b> References </b>
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4. Yamaguchi A, Katagiri T, Ikeda T, Wozney JM, Rosen V, Wang EA, Kahn AJ, Suda T, Yoshiki S (1991) . J Cell Biol 113:681–
4. Yamaguchi A, Katagiri T, Ikeda T, Wozney JM, Rosen V, Wang EA, Kahn AJ, Suda T, Yoshiki S (1991) . J Cell Biol 113:681–
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5. Nishimura, R, Kato Y, Chen D, Harris SE, Mundy GR, and Yoneda T (1998) J Biol Chem 273: 1872-1879
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6. Richard S, Torabi N, Franco GV, Tremblay GA, Chen T, et al. (2005). PLoS Genet 1(6): e74. doi:10.1371/journal.pgen.0010074
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Revision as of 13:09, 26 October 2010

Bone Formation
Background

The goal of this portion of the project was to synthetically differentiate stem cells into bone; this would act as the 'output' for our final system. Since stem cells can by tricky to genetically engineer, we chose to build our circuit in human endothelial kidney (HEK) cells. In the final system, HEK cells will secrete a diffusible morphogen to differentiate co-cultured stem cells. We used Bone Morphogenetic Protein 2 (BMP2) as our osteogenic signaling molecule; it is one of the central regulators of osteoblast differentiation in mammalian cells (1) and has been shown to induce transdifferentiation in multiple stem cell types (2, 3, 4). Here, we work with myoblastic progenitor (C2C12) and mesenchymal (C3HT101/2) stem cells, two cell lines which have been shown capable of osteoblast differentiation (5, 6). References

1. Hogan, B. L. (1996) Harvey Lect. 92, 83-98

2. Katagiri, T., Yamaguchi, A., Komaki, M., Ab, E., Takahashi, N., Ikeda, T., Rosen, V., Wozney, J. M., Fujisawa-Sehara, A., Suda, T. (1994) J. Cell Biol. 127, 1755-1766

3. Katagiri T, Yamaguchi A, Ikeda T, Yoshiki S, Wozney JM, Rosen V, Wang EA, Tanaka H, Omura S, Suda T (1990) . Biochem Biophy Res Commun 172:295–299

4. Yamaguchi A, Katagiri T, Ikeda T, Wozney JM, Rosen V, Wang EA, Kahn AJ, Suda T, Yoshiki S (1991) . J Cell Biol 113:681–

5. Nishimura, R, Kato Y, Chen D, Harris SE, Mundy GR, and Yoneda T (1998) J Biol Chem 273: 1872-1879

6. Richard S, Torabi N, Franco GV, Tremblay GA, Chen T, et al. (2005). PLoS Genet 1(6): e74. doi:10.1371/journal.pgen.0010074