PYRUVATE SUPPLEMENTATION ENHANCES VASCULAR ENDOTHELIAL GROWTH FACTOR PRODUCTION BY BONE MARROW-DERIVED MONONUCLEAR CELLS

• Published in: Japanese Journal of Transfusion and Cell Therapy Vol. 58; no. 1; pp. 26 - 32
• Main Authors: Kanno, Hitoshi, Iribe, Yuji, Aoki, Takako, Ogura, Hiromi, Fujii, Hisaichi
• Format: Journal Article
• Language: Japanese
• Published: The Japan Society of Transfusion Medicine and Cell Therapy 2012
• Subjects: critical limb ischemia; Endothelial progenitor cells; regenerative medicine; vascular endothelial growth factor
• ISSN: 1881-3011; 1883-0625
• DOI: 10.3925/jjtc.58.26

Bone marrow-derived mononuclear cells (BMMNC) include endothelial progenitor cells (EPC), which are characterized by their secretion of angiogenic factors such as vascular endothelial growth factor (VEGF) to recruit local endothelial cells, thereby enabling the establishment of new blood vessels. Implantation of BMMNC has been clinically used for therapeutic purposes in the treatment of critical limb ischemia (CLI); results showed that it was ineffective in a substantial number of cases. To evaluate the appropriate concentration of pyruvate to achieve the highest VEGF gene expression, cells were cultured with pyruvate at final concentrations up to 20mM in 5% CO2 for 2-4 days. The intracellular concentration of pyruvate was measured enzymatically and cell number and viability were determined. Expression levels of VEGF genes and numbers of CD31+/CD34+ cells were evaluated. Finally, VEGF levels in the conditioned medium were examined in each condition. Pyruvate supplementation in murine BMMNC cultures successfully increased intracellular pyruvate levels in a concentration-dependent manner, and 5mM pyruvate was found to be the most appropriate to maintain viable cell number and up-regulate VEGF gene after 2-day culture. In addition, VEGF in the conditioned medium was significantly elevated by the use of 5mM pyruvate after 4-day culture. From these results, we suggest that preconditioning of BMMNC with 5mM pyruvate for 2 days may be a useful way to safely and inexpensively enhance the angiogenic properties of BMMNC and the therapeutic effectiveness of cellular therapy for CLI.