Enhancement of organogenesis with small molecule drugs: Difference between revisions

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== Background ==
== Background ==
Vasculogenesis in vivo
'''Vasculogenesis in vivo'''
*formation of new blood vessels through the movement and differentiation of angioblasts (endothelial precursor cells)[insert picture from book]
*formation of new blood vessels through the movement and differentiation of angioblasts (endothelial precursor cells)[insert picture from book]
*induced by growth factors and extracellular matrix
*induced by growth factors and extracellular matrix
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*followed by angiogenesis (sprouting of new blood vessels from old '''existing''' ones) to form complete vascular tissue
*followed by angiogenesis (sprouting of new blood vessels from old '''existing''' ones) to form complete vascular tissue


 
'''
Necessity of vasculogenesis in vitro
Necessity of vasculogenesis in vitro'''
*current state of organogenesis
*current state of organogenesis
**possible to make thin patches of tissue
**possible to make thin patches of tissue

Revision as of 17:52, 13 May 2008

βββ -- using a bio-degradable scaffold. seed myocytes onto it. before scaffold breaks down, seed with angioblasts. use statins instead of VEGF to stimulate endothelial production. evaluate with fluoro microscopy.


Background

Vasculogenesis in vivo

  • formation of new blood vessels through the movement and differentiation of angioblasts (endothelial precursor cells)[insert picture from book]
  • induced by growth factors and extracellular matrix
  • during embryonic development
  • followed by angiogenesis (sprouting of new blood vessels from old existing ones) to form complete vascular tissue

Necessity of vasculogenesis in vitro

  • current state of organogenesis
    • possible to make thin patches of tissue
      • e.g. NASA researchers able to make small patch of cardiac tissue ~0.1 mm thick (but need 5 mm or so at least)
    • interior of patches have trouble obtaining enough oxygen.
  • solution: induce blood vessel formation via vasculogenesis during organogenesis

Research Goals

Project Details and Methods

1. Grow mesenchyme cells in a Wharton's Jelly-like solution (significantly containing hyaluronan). Hyaluronan is a significant component of the extracellular matrix and has been shown to contribute to tumor growth. In this step, we need to grow a layer of mesenchyme cells roughly five cells-thick. Hopefully the Jelly will promote this growth pattern.

(Several day's growth...)

2. Transfect the cells with Invitrogen™ Qtracker® 505 dye.

3. Apply statins to layers of cells to induce differentiation.

(Several day's growth and differentiation...)

4. Transfect the cells with Invitrogen™ Qtracker® 655 dye.

5. Quickly freeze sections of the cell layers to enable preparation of thin cross-sectional slices of tissue for observation. Then view under fluorescent microscope.

Predicted Outcomes

Resources

  • Researchers were able to grow rat ventricular cells in rotating bioreactors that quickly formed aggregates that rhythmically contracted in unison.
    • R.E. Akins, R.A. Boyce, M.L. Madonna, N.A. Schroedl, S.R. Gonda, T.A. McLaughlin, C.R. Hartzell. Tissue Engineering. April 1, 1999, 5(2): 103-118. doi:10.1089/ten.1999.5.103. [Article]
  • Asahara T, et al (1997). "Isolation of putative progenitor endothelial cells for angiogenesis." Science 275: 964-7. PMID 9020076. [Article]
  • Asahara T, et al (1999). "Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization." Circulation Research 85 (6): 221-8. PMID 10436164. [Article]
  • Hristov, et al (2003). "Endothelial progenitor cells: isolation and characterization." Trends in Cardiovascular Medicine 13 (5): 201-6. PMID 12837583. [Article]
  • Shaw J, et al (2004). "Hematopoietic stem cells and endothelial cell precursors express Tie-2, CD31 and CD45.". Blood Cells, Molecules, and Diseases 32 (1): 168-75. PMID 14757432. [Article]
  • Werner N, et al (2005). "Circulating Endothelial Progenitor Cells and Cardiovascular Outcomes.". New England Journal of Medicine 353: 999-1007. PMID 16148285. [Article]
  • A specific study of HMG-CoA reductase inhibitors (statins). The researchers found that the statins controlled hematopoietic progenitor cell differentiation.
    • Stefanie Dimmeler, Alexandra Aicher, Mariuca Vasa, Christiane Mildner-Rihm, Klaudia Adler, Michaela Tiemann, Hartmut Rütten, Stephan Fichtlscherer, Hans Martin and Andreas M. Zeiher. HMG-CoA reductase inhibitors (statins) increase endothelial progenitor cells via the PI 3-kinase/Akt pathway. J. Clin. Invest., 2001; 108(3): 391 - 397. [Article]
  • The Pax3 protein induces dense multi-layer aggregation of mammalian mesenchymal cells and facilitates the mesenchymal to endothelial transition.
    • Wiggan O'Neil, Mark Fadel, and Paul Hamel. Pax3 induces cell aggregation and regulates phenotypic mesenchymal-epithelial interconversion. Journal of Cell Science 115, 517-529 (2002) [Article]
  • Pax3 protein production in E. coli.
    • Stéphane C. Boutet, Marie-Hélène Disatnik, Lauren S. Chan, Kevin Iori and Thomas A. Rando. Regulation of Pax3 by Proteasomal Degradation of Monoubiquitinated Protein in Skeletal Muscle Progenitors. Cell 130, 342-362 (2007) [Article]

People

Steps (not for final page)

  • looking at cells in layers
  • scaffolds
  • statins and other signals for vasculogensis
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