Biological characteristics of synovial-derived mesenchymal stem cells in serum-free culture media and in vitro construction of three-dimensional cartilage in vitro
2017
Objective
To explore the biological characteristics of synovial fluid-derived mesenchymal stem cells (SF-MSCs) cultured in serum-free medium and the ability of in vitro reconstruction of three-dimensional cartilage combined with scaffold material.
Methods
Human SF-MSCs were cultured in serum medium and mesenchymal stem cells medium-serum free (MSCM-sf) respectively, then the proliferative ability and morphology of SF-MSCs were compared; The third passage SF-MSCs cultured in MSCM-sf were identified by flow cytometry, three-way(chondrogenic, osteogenic, adipogenic)differentiation assay and induced for chondrogenic differentiation when combined with polyglycolic acid/polylactic acid (PGA/PLA).
Results
SF-MSCs cultured in MSCM-sf had better morphology and proliferative ability than that cultured in serum medium. The expression levels of positive markers of the third passage SF-MSCs cultured in MSCM-sf, such as CD73 (99.5%), CD90 (98.9%) and CD105 (96.5%), were more than 95%. However, the overall negative markers (CD34, HLA-DR and CD11b) expressed less than 2%. Three-way differentiation staining was positive. The combination of SF-MSCs and PGA / PLA can be induced into cartilage in vitro.
Conclusions
SF-MSCs cultured in MSCM-sf can be amplified under the condition of maintaining the stem cell characteristics, and can be combined with PGA/PLA scaffold to construct three-dimensional cartilage in vitro.
Key words:
Mesenchymal stem cells; Synovial fluid; Culture media, serum-free
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