Abstract: Objective To compare the characteristics and chondrogenesis between CD204(-) ADSCs and CD204(+) ADSCs in vitro.Methods ADSCs were isolated and cultured,then the CD204(+) ADSCs and CD204(-) ADSCs were sorted by flow cell sorting.After expansion to passage 2,the morphology of CD204(+) and CD204(-) cells were observed in vitro.Adipogenic and osteogenic lineage differentiation in two cell groups were assessed by Oil Red O and alizarin red staining.Meanwhile,the two cell groups were cultured in chondrogenic media forming cell pellet and assessed by HE,Safranin O and collagen II staining.Results The expression of CD204 in adherent cells reached about 10% by FACS.Both of them all demonstrated fibroblast-like morphology and CD204(-) cells showed better proliferation ability.After adipogenic induction for 10 days,large amount of lipid-filled droplets were observed in two cell groups by Oil Red O staining,but CD204(+) cells demonstrated a better adipogenic ability.After osteogenic induction for 2 weeks,stronger positive staining was observed in CD204(-) cell group by alizarin red staining.After chondrogenic induction,the pellets formed by CD204(-) cells had a transparent white cartilage-like morphology while the CD204(+) cell pellets had a yellowish appreance.Histologically,the CD204(-) group showed a homogeneous cartilage-like structure with typical lacuna,strong safranin-O staining and strong collagen II staining,but pellets of CD204(+) group were almost fibrosis.Conclusion CD204(-) cells have the same characteristics with stem cells and high chondrogenic potential.CD204(-) cells could be an ideal seed cells in cartilage tissue engineering.

Key words: CD204, Adipose-derived stem cells, Chondrogenic potential, Cell sorting