International Journal of Stem Cells

Korean Society for Stem Cell Research (한국줄기세포학회)
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Mimicking the Human Articular Joint with In Vitro Model of Neurons-Synoviocytes Co-Culture

  • Jakub Chwastek (Department of Neurochemistry, Maj Institute of Pharmacology, Polish Academy of Sciences) ;
  • Marta Kedziora (Department of Neurochemistry, Maj Institute of Pharmacology, Polish Academy of Sciences) ;
  • Malgorzata Borczyk (Laboratory of Pharmacogenomics, Department of Molecular Pharmacology, Maj Institute of Pharmacology, Polish Academy of Sciences) ;
  • Michal Korostynski (Laboratory of Pharmacogenomics, Department of Molecular Pharmacology, Maj Institute of Pharmacology, Polish Academy of Sciences) ;
  • Katarzyna Starowicz (Department of Neurochemistry, Maj Institute of Pharmacology, Polish Academy of Sciences)
  • Received : 2023.04.12
  • Accepted : 2023.08.29
  • Published : 2024.02.28
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Abstract

The development of in vitro models is essential in modern science due to the need for experiments using human material and the reduction in the number of laboratory animals. The complexity of the interactions that occur in living organisms requires improvements in the monolayer cultures. In the work presented here, neuroepithelial stem (NES) cells were differentiated into peripheral-like neurons (PLN) and the phenotype of the cells was confirmed at the genetic and protein levels. Then RNA-seq method was used to investigate how stimulation with pro-inflammatory factors such as LPS and IFN𝛾 affects the expression of genes involved in the immune response in human fibroblast-like synoviocytes (HFLS). HFLS were then cultured on semi-permeable membrane inserts, and after 24 hours of pro-inflammatory stimulation, the levels of cytokines secretion into the medium were checked. Inserts with stimulated HFLS were introduced into the PLN culture, and by measuring secreted ATP, an increase in cell activity was found in the system. The method used mimics the condition that occurs in the joint during inflammation, as observed in the development of diseases such as rheumatoid arthritis (RA) or osteoarthritis (OA). In addition, the system used can be easily modified to simulate the interaction of peripheral neurons with other cell types.

Keywords

Acknowledgement

This research was funded by National Science Centre, Poland by grants OPUS (no. 2016/23/B/NZ7/01143), MINIATURA (no. 2019/03/X/NZ7/00644) and by Maj Institute of Pharmacology, Polish Academy of Sciences with Departmental Statutory funds. We would like to thank Galen Ortopedia Sp. z o.o.'s medical doctors and Department of Science, Innovation and Development for providing synovial membranes. Neuroepithelial stem cells were a kind gift of Anna Falk (Karolinska Institutet, Solna, Sweden).

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