Master Assignments

Affecting cellular responses to stimuli using growth factor sinks

Affecting cellular responses to stimuli using growth factor sinks

Osteoarthritis is associated with synovitis. Several pro-inflammatory cytokines are important mediators of inflammation and articular cartilage destruction, supporting a potential possibility of anticytokine therapy in these diseases. Several clinical trials have already demonstrated positive outcomes over disease progression. Although these treatments are very attractive, they are associated with limited efficacy because of the rapid clearance of antibodies by the synovium. A solution to overcome this problem is using microgels as a substrate to protect and extend the action of the antibodies. Dextran-tyramine (dexTA) and hyaluronic acid-tyramine (HA), are biocompatible polysaccharides, and HA is a natural component of the extracellular matrix of articular cartilage. Therefore, polymeric microgels composed of dexTA/dexHATA represent a good candidate for IA cytokine capturing.

In this assignment you are going to stimulate cells with varying growth factors, and measure their response. Subsequently you are going to capture these growth factors with microgels and see if the cellular response changes.

Main question
Is it possible to capture cytokines using microgels functionalized with VHH antibodies from a cell culture, and does this result in a biological response?

Sub questions

  1. Is it possible to capture BMP-7 and measure the change in ALP production/luciferase expression in C2C12 cells?
  2. Do the recombinant proteins of FRZB and DKK stimulate C2C12 cells in their osteogenic differentiation?
  3. Do the VHH cross-react with rat and mouse FRZ and DKK?
  4. Is it possible to capture FRZB and DKK-1 from a stimulated cell culture?
  5. Is it possible to co-capture FRZB and DKK-1 from a stimulated cell culture?

Techniques (in between brackets is for master students only)

  • (Production of microfluidic chips)
  • (Production of biotinylated gels)
  • (Functionalization of biotinylated gels)
  • C2C12 cell culture
  • ALP-assay
  • ELISA for cross-reactivity

Daily supervisor: Ir. Lisanne Karbaat

If you are interested in this assignment, please contact: Dr. Janneke Alers, contact person for all DBE assignments