Isolation and Characterization of Multipotential Mesenchymal Cells from the Mouse Synovium

Abstract

The human synovium contains mesenchymal stem cells (MSCs), which are multipotential non-hematopoietic progenitor cells that can differentiate into a variety of mesenchymal lineages and they may therefore be a candidate cell source for tissue repair. However, the molecular mechanisms by which this can occur are still largely unknown. Mouse primary cell culture enables us to investigate the molecular mechanisms underlying various phenomena because it allows for relatively easy gene manipulation, which is indispensable for the molecular analysis. However, mouse synovial mesenchymal cells (SMCs) have not been established, although rabbit, cow, and rat SMCs are available, in addition to human MSCs. The aim of this study was to establish methods to harvest the synovium and to isolate and culture primary SMCs from mice. As the mouse SMCs were not able to be harvested and isolated using the same protocol for human, rat and rabbit SMCs, the protocol for humans was modified for SMCs from the Balb/c mouse knee joint. The mouse SMCs obtained showed superior proliferative potential, growth kinetics and colony formation compared to cells derived from muscle and bone marrow. They expressed PDGFRá and Sca-1 detected by flow cytometry, and showed an osteogenic, adipogenic and chondrogenic potential similar or superior to the cells derived from muscle and bone marrow by demonstrating in vitro osteogenesis, adipogenesis and chondrogenesis. In conclusion, we established a primary mouse synovial cell culture method. The cells derived from the mouse synovium demonstrated both the ability to proliferate and multipotentiality similar or superior to the cells derived from muscle and bone marrow.

WNT-Inflammasome Signaling Mediates NOD2-Induced Development of Acute Arthritis in Mice

“…Synovial cells from mice were isolated, as previously described, with certain modifications (26). Briefly, mice were positioned such that the knee joints were exposed by a midline skin incision.…”

Cell-based articular cartilage repair: the link between development and regeneration

“…Subsequent studies revealed that human synovial MSCs have greater expansion and chondrogenic ability in vitro than MSCs from bone marrow, periosteum, muscle, and adipose tissue 77 . The weight of cartilaginous pellets from cultured mouse synovial MSCs is significantly greater than that from cultured bone marrow MSCs 68 . Extracellular matrix deposited by synovial MSCs delays replicative senescent chondrocyte dedifferentiation and enhances redifferentiation 73 .…”

“…The chondrogenic potential of synovium-derived MSCs and their application in AC repair have been studied in vitro and in vivo 68–75 . An early in vitro study demonstrated that human multipotent MSCs can be isolated from the synovial membrane of knee joints.…”

Mouse synovial mesenchymal stem cells increase in yield with knee inflammation

“…Ten IFPs were also harvested from untreated knees as a control group. Ten IFPs were minced and digested in a 1 mg/ml collagenase (Wako, Osaka, Japan) for 15 min and passed through a 40-μm filter (Becton Dickinson, Franklin Lakes, NJ) 15. Dissociated cells and undigested IFPs were cultured together in a culture dish of 10-cm 2 (Nalge Nunc International, Rochester, NY) in Dulbecco's modified Eagle's medium (DMEM: Wako) containing 10% fetal bovine serum (FBS: Invitrogen, Carlsbad, CA), 100 U/ml penicillin and 100 mg/ml streptomycin (Invitrogen) and 5 ng/ml recombinant human FGF-2 (BioVision, Milpitas, CA) at 37°C with 5% humidified CO 2 .…”