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Characterization of the response of encapsulated MSCs to inflammatory stimuli

Characterization of the response of encapsulated MSCs to inflammatory stimuli

Osteoarthritis is associated with - and exacerbated by - inflammation. Although clinically anti-inflammatory drugs are used, they are largely ineffective due to their short residence time. We are exploring an exciting novel way to decrease the inflammation in the knee joint: intra-injection of MSCs that secrete anti-inflammatory factors. Like the drugs, MSC treatment efficacy is limited by its rapid clearance from our joints. We pioneer a novel method to prevent this rapid clearance: Microfluidic encapsulation of MSCs in micrometer sized hydrogel matrices. However, the effect of encapsulation on MSCs anti-inflammatory properties has remained unknown. Therefore this study aims to determine the effects of biomaterial characteristics and composition on anti-inflammatory performance.

In this assignment you will encapsulate MSCs using a chip-based microfluidic droplet generator. Encapsulated MSCs will be stimulated with varying cytokines. Using this approach, you will determine if encapsulation and biomaterial composition has an effect on the MSCs therapeutic performance.

 





Main question

Do encapsulated MSCs respond differently to inflammatory stimuli change depending on their surrounding biomaterial?

Sub questions (in between brackets is for master students only)

  1. How do we get the highest RNA quality from encapsulated MSCs
  2. What is the response on RNA-level of non-encapsulated MSCs to inflammatory cytokines such as IL-1β, TNF-α and IFN-γ?
  3. What is the response on RNA-level of encapsulated MSCs to inflammatory cytokines such as IL-1β, TNF-α and IFN-γ?
  4. Does biomaterial composition and stiffness influence the therapeutic response of MSCs?
  5. (What is the response on protein level?)

Techniques (in between brackets is for master students only)

  • (Production of microfluidic chips)
  • Encapsulation of MSCs in dexTA and dexHATA microgels
  • MSC cell culture
  • MSC isolation from rat femurs
  • RNA isolation from encapsulated cells
  • RT-qPCR
  • (protein dot blot)


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