title: Isolation and Cellular Phenotyping of Mesenchymal Stem Cells Derived from Synovial Fluid and Bone Marrow of Minipigs.
abstract: Mesenchymal stem cells (MSCs) have been established after isolation from various tissue sources, including bone marrow and synovial fluid. Recently, synovial-fluid-derived MSCs were reported to have multi-lineage differentiation potential and immunomodulatory features, which indicates that these cells can be used for tissue engineering and systemic treatments. This study presents a protocol for simple and non-invasive isolation of MSCs derived from the bone marrow and synovial fluid of minipigs to analyze cell surface markers for cell phenotyping and in vitro culturing. Using sexually mature six-month-old minipigs, bone marrow was extracted from the iliac crest bone using a bone marrow extractor, and the synovial fluid was aspirated from the femorotibial joint. Procedures for the collection of samples from both sources were non-invasive. The protocols for effective isolation of MSCs from harvested cell sources and for creating in vitro culture conditions to expand stable MSCs from minipigs and the application of systemic autologous treatments are provided. For cell phenotyping, the cell surface markers of both cells were analyzed using flow cytometry. In the results, the MSCs were isolated from the synovial fluid of the minipigs and showed that synovial-fluid-derived MSCs have a similar morphology and cell phenotype to bone-marrow-derived MSCs. Therefore, non-invasively obtained synovial fluid is a valuable source of MSCs.
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University; PWG Genetics Pte Ltd.
authors.forename: Won-Jae
authors.initials: WJ
authors.lastname: Lee
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University.
authors.forename: Ji-Sung
authors.initials: JS
authors.lastname: Park
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University.
authors.forename: Si-Jung
authors.initials: SJ
authors.lastname: Jang
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University.
authors.forename: Seung-Chan
authors.initials: SC
authors.lastname: Lee
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University.
authors.forename: HyeonJeong
authors.initials: H
authors.lastname: Lee
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University.
authors.forename: Jae-Hoon
authors.initials: JH
authors.lastname: Lee
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University.
authors.forename: Gyu-Jin
authors.initials: GJ
authors.lastname: Rho
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University; sllee@gnu.ac.kr.
authors.forename: Sung-Lim
authors.initials: SL
authors.lastname: Lee
mesh_terms: D000818:Animals; D001853:Bone Marrow; D001854:Bone Marrow Cells; D002454:Cell Differentiation; D059630:Mesenchymal Stem Cells; D013552:Swine; D013556:Swine, Miniature; D013582:Synovial Fluid