Earticle

초록

영어

Current studies have revealed the capacity of mesenchymal stem cells (MSCs) in term of immunomodulatory properties, and this distinct potential is downgraded according to the disease duration of patients-derived MSCs. In order to enhance the immunomodulatory and anti-tumorigenic properties of the rheumatoid arthritis (RA) joints-derived MSCs, we aggregate synovial fluid-derived MSCs from RA joints (RA-hMSCs) into 3D-spheroids by the use of hanging drop culture method. Cells were isolated from synovial fluids of RA joints with longstanding active status over 13 years. For aggregation of RA-hMSCs into 3D-spheroids, cells were plated in hanging drops in 30 μL of advanced DMEM (ADMEM) containing 25,000-30,000 cells/ drop and cultured for 48 h. To analyze the comparative immunomodulatory effects of 3D-spheroid and 2D monolayer cultured RA-hMSCs and then cells were cultured in ADMEM supplemented with 20% of synovial fluids of RA patients for 48 h and were evaluated by qRT-PCR for their expression of mRNA levels of inflammatory and antiinflammatory markers. Cellular aggregation of RA-hMSCs was observed and cells were aggregate into a single sphere. Following treatment of RA patient’s synovial fluids into the RA-hMSCs, spheroids formed RA-hMSCs showed significantly (p < 0.05) higher expression of TNFα stimulated gene/protein 6 (TSG-6) than the monolayer cultured RAhMSCs. Therefore, the 3D-spheroid culture methods of RA-hMSCs were more effective than 2D monolayer cultures in suppressing inflammatory response treated with 20% of RA-synovial fluids by expression of TNFα (TSG-6) according to the immune response and enhanced secretion of inflammatory factors.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Characterization of RA-hMSCs prior to 2D- or 3D-culture method
Cell proliferation assay
Spheroid generation
Synovial fluid treatment
Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis
Statistical analysis
RESULTS
Cell surface markers for MSCs
Multilineage differentiation capacity of MSCs
Morphology change during RA-SF treatment culture of RA-hMSCs
Expression mRNA levels of anti-inflammatory and antitumorigenic molecules
DISCUSSION
REFERENCES

키워드

저자

• Seung-Chan Lee [ Apures, Inc., Central Research Center, Pyeongtaek 17792, Korea ]
• Chae-Yeon Hong [ College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea ]
• Yong-Ho Choe [ College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea ]
• Tae-Seok Kim [ College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea ]
• Won-Jae Lee [ College of Veterinary Medicine, Kyungpook National University, Daegu 41566, Korea ]
• Gyu-Jin Rho [ College of Veterinary Medicine, Gyeongsang National University, Research Institute of Life Sciences, Gyeongsang National University, Jinju 52828, Korea ]
• Sung-Lim Lee [ College of Veterinary Medicine, Gyeongsang National University, Research Institute of Life Sciences, Gyeongsang National University, Jinju 52828, Korea ] Corresponding Author

참고문헌

발행기관

간행물

관심저널 등록

• 간행물명

  Journal of Animal Reproduction and Biotechnology

• 간기

  계간

• pISSN

  2671-4639

• eISSN

  2671-4663

• 수록기간

  2019~2026

• 십진분류

  KDC 527 DDC 636