In vitro culture of autologous mesenchymal stem cells from the joint drainage fluid after knee arthroscopy: a feasibility study / 中国组织工程研究

ABSTRACT

BACKGROUND:Mesenchymal stem cells have a extreme prospect in orthopedics, which show great potential especially in the treatment of articular cartilage defect disease. Bone marrow is the main source of mesenchymal stem cells, and the iliac puncture is a conventional way to obtain bone marrow, but is restricted by the limited resources and strict technical requirements. Therefore, it is of great significance to explore new effective and convenient sources of mesenchymal stem cells. OBJECTIVE:To explore the feasibility of autologous mesenchymal stem cells derived from the joint drainage fluid after knee arthroscopy.METHODS: We selected eight patients who underwent arthroscopic surgery to collect joint drainage fluid by pre-made sterile blood bag before the wound closure. Precipitation with hydroxyethyl starch and density gradient centrifugation method were performed to isolate and culture mesenchymal stem cells from the joint drainage fluid. Cell morphology, growth curve, surface marker identification were observed and detected using flow cytometry. Then, adipogenic, chondrogenic and osteogenic differentiation of cells were induced and identified by oil red O, toluidine blue staining, and alizarin red staining, respectively. RESULTS AND CONCLUSION:The cultured cells were spindle-shaped, adherently grew and had good proliferation ability, which were positive for CD44, CD90, CD105 and CD73, but not for CD45. Under standard inductions, the cultured cells were induced to differentiate into osteoblasts, adipocytes and chondrocytes. Therefore, these cells were confirmed as mesenchymal stem cells. Mesenchymal stem cells were successfully isolated from the joint drainage fluid of eight patients and had no difference in cell morphology, proliferation and phenotypes. To conclude, the joint drainage fluid is an ideal source of mesenchymal stem cells with the guaranteed quality and quantity.