Suggs:Meeting Notes 10/28/05

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Suggs Lab

Basement


Meta

  • TGF-B (sticky) -> SMCs differentiate
  • PDGF-BB (not sticky) -> recruit SMC progenitors associate with endothelial cells -> cleaves TGF-B off site -> SMCs.
    • In vivo, primary function seems to be contained in a matrix and then liberated when needed.
  • Can this recruit another population of cells to form SMCs?
  • Alt-Ester-Amide links. Walk a line of protein and synthetic polymer, can form secondary structure (i.e. a helix). Need doublet/triplet specificity -> polyionic gels, enzyme susceptability, GF binding, integrin binding. Commercially viable/producable pov.
  • Side chains dictate everything (degrading).
    • Which ones to pick?
      • Evaluate structure of polymer, and behavior with other proteins, cells, etc.


Mena

  • Linear monomer (100 mg) -> cyclic monomer
  • Fabricate linear monomer prior to cyclization
    • Triethyl Amine in DMF. Heat linear monomer in anhydrous environment to make cyclic.
  • Cyclic monomer
    • Purification protocol from patent (technical difficulties)
      • Water contamination
      • N2 run-out
    • Continue with 2nd batch.
    • Crumbly vs. Hairy? Physical precipitate, no worries.


Scott

  • Fibrin gels ~ 2 wks.
    • LD gel, add inhibitors to prohibit degradation = toxic to cells.
    • Bulk degradation
      • Mechanical loss (gels become softer)
      • Loss of mass = cells liberated.
    • Weight loss first. Rheometers.
  • Species of thrombin
    • Porcine fibrin + bovine thrombine [100u 2 day decay, 50u 3 day decay]
      • 8 days, no 100u, some 50u
    • Porcine fibrin + human thrombin
    • Fibrin -> mechanical cross-link
    • Thrombin -> coagulation protein - fibrinogen -> fibrin
  • Tabulate ranges of fibrinogen and thrombin.
  • Back off on thrombin!!!


Charlie

  • TGF-B from fibrin gels, (new literature) stimulate actin/myosin for smooth muscle cell differentiation.
  • Release kinetics.
  • Differentiation protocols = 1-2 weeks.
  • During mass loss, TGF-B dump.
  • Goal: Constant release over full time, then go away.
  • Proteonomics. Protein expression in gels.


Office Space

  • Charlie, Jessica upstairs.
  • Scott stays in Basement.
  • Computers move to old-Charlie space.
    • Free space around Mena area.
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