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Author Spotlight: Isolation and Culture of Primary Synovial Macrophages and Fibroblasts from Murine Arthritis Tissue
Published on: February 24, 2023
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Multilineage-differentiating stress-enduring (Muse)-like cells exist in synovial tissue
Eriko Toyoda1, Masato Sato1, Takumi Takahashi1
1Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan.
Regenerative Therapy
|December 8, 2018
Summary
Multipotent adult stem cells, known as Muse-like cells, can be isolated from human synovial membranes, offering a promising alternative for cartilage regeneration. These cells, even from older patients, demonstrate significant chondrogenic potential, potentially overcoming limitations of current regenerative medicine approaches.
Area of Science:
- Regenerative Medicine
- Stem Cell Biology
- Orthopedic Research
Background:
- Cartilage regeneration is crucial for treating cartilage defects and restoring joint function.
- Limited availability of autologous chondrocytes hinders clinical application.
- Investigating alternative cell sources like synovial membrane-derived cells is essential.
Purpose of the Study:
- To determine the existence and chondrogenic potential of synovial membrane-derived multilineage-differentiating stress-enduring (Muse)-like cells.
- To evaluate Muse-like cells as a viable alternative cell source for cartilage regeneration.
- To compare the chondrogenic potential of Muse-like cells with unsorted synovial cells.
Main Methods:
- Isolation of stage-specific embryonic antigen-3 (SSEA-3)-positive cells from human synovial membranes via fluorescence-activated cell sorting.
- Culture of SSEA-3-positive cells in methylcellulose to form Muse clusters and subsequent expansion.
- Assessment of chondrogenic potential using pellet culture systems, evaluating extracellular matrix production and specific staining (proteoglycan, safranin O, toluidine blue, type II collagen).
Main Results:
- Synovial membranes yielded SSEA-3-positive cells exhibiting Muse-like characteristics, including cluster formation and expression of key stem cell markers (NANOG, OCT3/4, SOX2).
- M-cluster-derived mesenchymal stem cells (MSCs) demonstrated increased extracellular matrix production, positive staining for proteoglycans and type II collagen, indicating successful chondrogenesis.
- Unsorted synovial cells also formed clusters with comparable chondrogenic potential to Muse-like cells, suggesting a practical alternative.
Conclusions:
- Muse-like cells are isolatable from human synovial membranes, even in elderly patients, providing a valuable source for regenerative medicine.
- The cluster-forming cell population within synovial cells possesses significant chondrogenic potential.
- These findings suggest a more practical and accessible cell source for cartilage regeneration therapies.
Keywords:
APC, allophycocyaninBSA, bovine serum albuminCartilageChondrogenic potentialDAB, 3,3′-diaminobenzidineDAPI, 4′,6-diamidino-2-phenylindoleFBS, fetal bovine serumFITC, fluorescein isothiocyanateHRP, horseradish peroxidaseIg, immunoglobulinM-cluster, cluster cultured from synovial–Muse cellsMC, methylcelluloseMSC, mesenchymal stem cellMultilineage-differentiating stress-enduring cellsMuse, multilineage-differentiating stress-enduring cellsPBS, phosphate-buffered salinePE, phycoerythrinRegenerative medicineSSEA-3, stage-specific embryonic antigen-3SY-cluster, cluster cultured from unsorted synovial cellsStage-specific embryonic antigens-3αMEM, alpha-minimum essential medium
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