Impact of vascular endothelial growth factor and basic fibroblast growth factor on culture of rat peripheral blood progenitor cells
10.3760/cma.j.issn.1001-7097.2010.12.009
- VernacularTitle:血管内皮生长因子和碱性成纤维细胞生长因子对大鼠外周血内皮祖细胞培养的影响
- Author:
Jianjiang ZHANG
;
Liping ZHENG
;
Hua WANG
- Publication Type:Journal Article
- Keywords:
Vascular endothelial growth factor A;
Fibroblast growth factor 2;
Stem cells;
Fibronectins
- From:
Chinese Journal of Nephrology
2010;26(12):915-919
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the impact of different culture conditions on the growth of the endothelial progenitor cells(EPCs) from rat peripheral blood. Methods Mononuclear cells obtained from rat peripheral blood were isolated by using density gradient centrifugation. According to the culture medium added with vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and the dishes were precoated with fibronectin or not, the mononuclear cells were divided into different groups and cultured under different conditions in vitro to compare the differences of the growth conditions. The different growth conditions were recorded and the result was calculated by statistics software. At last, the cells were identified by immunohistochemistry and immunofluorescence. Results The mononuclear cells from rat peripheral blood grew in the manner of keeping close to the wall in vitro. The cells and cell colonies cultured for 7 days hinted:under the same culture conditions, precoating FN was beneficial to the adherent proliferation of EPCs (t=4.43, P＜0.05; t=3.70, P＜0.05). Excluding the impact of fibronectin, growth factors could promote mononuclear cells differentiation to EPCs, indicating that growth factors enhanced proliferation of EPCs (t =-13.22, P＜0.01; t =-10.96, P＜0.01). Immunohistochemistry and immunofluorescence showed that, at day 4, 7, 10 of cells cultured, CD34 and CD133 expressions were increased gradually [(35.7±4.2)%, (60.1±3.8)%, (81.8±6.4)%; (3.2±0.9)%, (18.4±7.3)%,(32±3.8)%, respectively]; at day 14, both were decreased [(32.1±5.4)%, (1.9±2.7)%]; but Flk-1was increased at day 4, 7,10, 14 [(31.2±3.5)%, (40.6±5.3)%, (71.2±8.4)%, (81.5±4.1)%].Conclusions Fibronectin is conducive to adhesion and proliferation of EPCs. VEGF and bFGF play an important role in the differentiation of EPCs. The success culture of EPCs in vitro will provide a sufficient number of seed cells for its application in vascular tissue engineering, and offer new ideas for peripheral blood stem cell transplantation for the treatment of various diseases.
