Studies On Isolation And Identification Of Human Cartilage-derived Pluripotent Stem Cells

We have isolated and identified a novel stem cell population from articular cartilage of naturally aborted human fetus of 8-12 weeks,and named human cartilage-derived pluripotent stem cells(hCPSCs).After a two-step digestion with trypsin and type ?collagenase,primary and passage cells were cultured in a H-DMEM followed by knock-out DMEM.Cell cycle analyses of passage 3 and passage 10 cells indicated that these cells were relatively primary,undifferentiated,and mostly in G0/G1 phase.hCPSCs demonstrated an expression pattern of biomarkers of human embryonic stem cells(OCT4,NANOG,Sox2,Ssea-3 and Ssea-4),as well as human adult stem cells(CD29,CD44,CD90,CD73 and CD 10)analyzed by flow cytometry.These cells expressed hESC-related genes(OCT4,NANOG,TDGF-1)and mesoderm-related gene(TBX5)but not hTERT or genes associated with other germ layers such as MEF2C,SOX17,NEUROD1,or PAX6 examined by reverse transcriptase-polymerase chain reaction analyses.These results suggested that they are a group of stem cell population with stemness in-between human embryonic stem cells and human adult stem cells.After adding inducing factors,hCPSCs were able to differentiate into adipocyte-like cells,osteoblast-like cells,chondrocyte-like cells,neuron-like cells,and islet-like cells,suggesting they were able to differentiate toward cells types of all three primitive germ layers similar to that of human embryonic stem cells.Our findings may provide research bases and a potential stem cell population for cell therapy in the treatment of T1D.