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. Pro-inflammatory cytokines are important mediators of inflammation and articular cartilage destruction. Indeed, anti-cytokine therapy has great potential to treat this disease. Several clinical trials have already demonstrated positive outcomes over disease progression. Although these treatments are very attractive, they are associated with limited efficacy due to their rapid clearance of antibodies by the synovium. We propose to overcome this challenge by endowing micrometer sized hydrogel (microgels) with cytokine capturing antibodies. Importantly, the microgels cannot be rapidly cleared from the joint and thus allow the antibodies to perform their anti-cytokine function for a substantially prolonged period of time. To create the microgels, we will use advanced microfluidic droplet generators and state-of-the-art enzymatically crosslinking biomaterials.

In this assignment you are going to fabricate antibody-modified microgels using microfluidic droplet generation. Subsequently you are going to determine the efficacy of the microgels to capture these cytokines.

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

Sub questions

  1. What is the response on RNA-level of MSCs to IL-1β, TNF-α and IFN-γ?
  2. Is it possible to capture TNF-α from a medium using clinically used antibodies?
  3. Is it possible to influence the cellular response to TNF-α on a RNA level by using TNF-α neutralizing antibodies?
    For master students only
  4. Is it possible to produce microgels functionalized with antibodies?
  5. Do  coupled antibodies have same effect as non-coupled antibodies?
  6. Once all in vitro work is done, develop a ‘’werkprotocol’’ for the animal experiments which are to be conducted with these functionalized microgels.

Techniques

  • (Production of microfluidic chips)
  • (Production of biotinylated gels)
  • Functionalization of biotinylated gels
  • Cell culture
  • Biochemical assays such as ALP-assay
  • Viability and metabolic assays

Gene expression analysis using RT-qPCR

Supervision
L.P. Karbaat (Lisanne)
PhD Candidate
Supervision
dr. J.C.H. Leijten (Jeroen)
Assistant Professor
Contact person
dr. J.C. Alers (Janneke)
Lecturer