Differentiation of C57/BL6 mice bone marrow mononuclear cells into early endothelial progenitors cells in different culture conditions

Author(s)

Carneiro, Giane D. ^[1] ; Godoy, Juliana A. P. ^[1] ; Werneck, Claudio C. ^[2] ; Vicente, Cristina P. ^[1]

Affiliation

^[1] Univ Estadual Campinas, Dept Struct & Funct Biol, UNICAMP, Sao Paulo - Brazil

^[2] Univ Estadual Campinas, Inst Biol, UNICAMP, Dept Biochem & Tissue Biol, Sao Paulo - Brazil

• Document type: Journal article
• Source: Cell Biology International; v. 39, n. 10, p. 1138-1150, OCT 2015.
• Web of Science Citations: 5

Abstract

Endothelial progenitor cells (EPCs) can be isolated from bone marrow and characterized by the expression of cellular markers such as CD34, CD133, VEGFR2, CD31, and VE-Cadherin, by the uptake of acetylated low-density lipoprotein and by in vitro tube formation in tridimensional matrices. These cells are able to differentiate into mature endothelial cells and participate in the re-endothelization of damaged vessels. In this work, we tested different cultured media that can promote the proliferation and differentiation of mononuclear cells (MNCs) into early EPCs, with defined concentrations of growth factors and serum in order to establish a composition that may ensure us the reproducibility of our cultures. MNCs from mice bone marrow were cultivated using selective culture media containing DMEM or M199 supplemented with 10% FBS, VEGF, bFGF, and IGF, for 3, 7, and 14 days. Differentiation into early EPCs was analyzed using immunohistochemistry, FACS and western blotting and by functional parameters as uptake of ac-LDL, and formation of vessel-like structures. The cells cultivated with medium DMEM-M1 (DMEM plus VEGF, bFGF and IGF) expressed CD34, CD133, CD31, VEGFR2, and VE-Cadherin at all culture time-points with increased expression of these markers after 7 days. Only EPCs cultured for 30 days were able to form vessel-like structure. The uptake of ac-LDL was observed after 3, 7, 14, and 30 days, confirming the differentiation of mononuclear cells into early EPCs. DMEM-M1 was able to sustain MNCs proliferation and differentiation, increasing the expression of the characteristic EPC markers, allowing the expansion of early EPCs in culture in a similar way to that observed in commercial available media. (AU)

• FAPESP's process: 10/19916-7 - Evaluation on elastic fiber synthesis in cell culture obtained from fibrillin-1 defficient mice: Losartan's effect studies
• Grantee: Claudio Chrysostomo Werneck
• Support Opportunities: Regular Research Grants
• FAPESP's process: 12/23640-2 - The role of endothelial progenitor cells in arterial thrombosis and vascular remodeling observing the interference of dermatan sulfate, an antithrombotic glycosaminoglycan in this process
• Grantee: Cristina Pontes Vicente
• Support Opportunities: Regular Research Grants