Tc14012

Primary bone marrow-derived human MSCs were purchased from Sigma-Aldrich. MSCs were cultured in basic medium (α-MEM; Gibco) supplemented with 10% fetal bovine serum (FBS; Gibco), 100 U/ml penicillin (Gibco), 10 μg/ml streptomycin (Gibco), 2 mM l-glutamine (Gibco), and 0.2 mM l-ascorbic acid 2-phosphate magnesium salt (ASAp, Sigma-Aldrich) at 37°C in a humid atmosphere with 5% CO₂. Human monocytic THP-1 cells were maintained in culture in Roswell Park Memorial Institute medium (RPMI 1640; Invitrogen) culture medium containing 10% heat-inactivated FBS (Invitrogen) and supplemented with 10 mM HEPES (Gibco), 1 mM pyruvate (Gibco), 2.5 g/l d-glucose (Merck), and 50 pM ß-mercaptoethanol (Gibco). THP-1 monocytes are differentiated into M1 macrophages by 24 h incubation with 150 nM phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich) followed by 24 h incubation in RPMI medium containing 20 ng/ml of IFN-γ (R&D Systems) and 10 pg/ml of lipopolysaccharides (LPS; Sigma-Aldrich). Jurkat-T cells were maintained in RPMI 1640 (Invitrogen) supplemented with 10% heat-inactivated FBS, 100 U/ml penicillin, and 100 mg/ml streptomycin (Gibco) at 37°C and 5% CO₂. For MSC priming, MSCs were incubated with the media containing CCX771 (100 nM; ChemoCentryx) or TC14012 (30 μM; Sigma-Aldrich) for 48 h and collected for cell transplantation.

Recombinant human CXCL12 was obtained from R&D Systems (Wiesbaden, Germany). Calcein-AM was purchased from Dojindo (Dojindo Laboratories, China). Polyclonal rabbit anti-CXCR4 and polyclonal rabbit anti-CXCR7 were obtained from Abcam (Cambridge, MA, USA). Polyclonal rabbit anti-β-Actin antibody, TC14012 and AMD3100, were obtained from Sigma (Deisenhofen, Germany). CCX771 was obtained from ChemoCentryx (Mountain View, CA, USA).

Osteogenic and chondrogenic differentiation media were purchased from Lonza. TC14012 was obtained from Sigma-Aldrich. CCX771 was a kind gift from Dr. Mark Penfold (ChemoCentryx).