PHYSIOLOGICAL ROLE OF BONE MARROW ADULT STEM CELL CD133+

Abstract

Objective: Тo analyze the in vitro comportment of the CD133+ cells and their profile for cytokines secretion. Methods: Bone marrow samples were obtained from 5 healthy individuals. Mesenchymal stem cells CD133+ extracted by magnetic bead from human bone marrow mononuclear cells (BMMNCs). BMMNCs were isolated by density-gradient centrifugation over Ficoll-400. Isolated CD133+ cells were plated on 0.2% gelatin-coated wells in the presence of free culture medium for cytokines analysis and with rich culture medium for spherical stem cell cluster generation. The proteins detected by protein array from the three independent cell preparations were considered as bioactive proteins. Results: Isolated CD133+ cells using magnetic bead present more than 87±6% and 8±5% CD34+ cells as assessed by flow cytometry and differentiate into adherent cells. CD133+ cells generate spheroid cell clusters in rich culture medium. Biological classification of the bioactive proteins secreted by the primo culture of CD133+/ CD34+ BMMNCs after 36h in conditioned culture medium showed the presence of several categories of cytokines. Among these, the cardiac hypertrophic factor, the pro-angiogenic factors, the pro-inflammatory factors, wound healing factors such as MMPs-TIMPs, the neurophilic factors, the morphogenetic proteins and hematopoietic growth factors can be mentioned. Conclusion: Our results indicate that CD133+ extracted from BMMNCs secretes important bioactive proteins. The multiple properties of these cytokines undoubtedly offer many therapeutic advantages.

Keywords

• cd133+ stem cells
• spheroids formation
• bone marrow
• cytokines
• growth factors.