Stem flow hmsc kit

ADMSCs were subjected to flow cytometry analyses for confirmation that ADMSCs maintain their phenotypic characteristics in vitro. After the third passage, cells were harvested, centrifuged, and resuspended in PBS at a minimum concentration of 1 × 10⁶ cells/ml. Immunophenotyping characterization of ADMSCs was performed with antibodies against the following combination of human antigens: CD11b, CD19, CD34, CD44, CD45, CD73, CD90, and CD105 (BD Stem Flow hMSC kit, BD Cat. No: 562245). Flow cytometry analyses were performed using Navios (Beckman Coulter, USA) before application to all patients in this study. The data were analyzed with KALUZA software (Beckman Coulter).

Sufficient cryopreserved ADMSC vials were thawed to provide the required dose for administration. The frozen ADMSCs were thawed and cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% human serum, 1% penicillin-streptomycin solution, and 1% stable glutamine (Biological Industries, Kibbutz Beit Haemek, Israel) at 37°C and 5% CO₂ incubator as described before [16 ]. Briefly, ADMSCs were recovered and resuspended in physiological saline solution and transferred to the patient with the temperature-controlled bag. The morphology of cell appearance and viability were assessed besides the identification of phenotypic characteristics of the cells and microbiological tests. Immunophenotyping characterization of ADMSC was performed by using flow cytometry analyses (Beckman Coulter, USA) with antibodies against CD45, CD73, CD90, CD105, CD11b, CD19, CD34, and CD44 (BD Stem Flow hMSC kit).