Establishment and Analysis of Three-Dimensional Angiogenesis Model of Endothelial Progenitor Cell from Peripheral Blood_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

SHI Sen ¹ , HE Yanzheng ¹ , LIU Yong ¹ , GE Hongwei ² , HU Bo ¹ , WEN Xuegang ³ , ZENG Hong ¹ , ZHONG Wu ¹ , YANG Hui ¹

1.Department of Vascular Surgery, Affiliated Hospital of Luzhou Medical College,Luzhou 646000, Sichuan Province, China;;
2.Department of Vascular Surgery, The First People’s Hospital of Changzhou City of Jiangsu Province, Changzhou213017, China;;
3.Department of Hepatobiliary Surgery, People’s Hospital of Zhongxian, Zhongxian 404300, Chongqing City, China Corresponding Author： HE Yan-zheng， E-mail： shisen80@163.com;

Corresponding author：

Keywords：

Endothelial progenitor cell; Three-dimensional angiogenesis model; Vascular endothelial growth factor; Rat tail collagen gel

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Objective To establish the three diamension-model and to observe the contribution of endothelial progenitor cell (EPC) in the angiogenesis and its biological features. MethodsEPC was obtained from the rats’ peripheral blood. Its cultivation and amplification in vitro were observed, and the function of the cultural EPC in vitro was detected. The three diamension-model was established and analyzed. ResultsEPC was obtained from the peripheral blood successfully. The proliferation of the EPC which induced with VEGF(experimental group) was better than that without VEGF （control group） at every different phase (P＜0.01). It was found that EPC grew into collagen-material from up and down in the three diamension-model, and its pullulation and infiltration into the collagen were seen on day 1 after cultivation. With the time flying, there were branch-like constructions which were vertical to the undersurface of collagen and interlaced to net each other. It showed that in experimental group the EPC grew fast, its infiltration and pullulation also were fast, the branch-like construction was thick. But in control group, the EPC grew slowly, infiltration and pullulation were slow, the branch-like construction was tiny and the depth of infiltration into collagen was superficial. The number of new vessels in experimental group was larger than that in the control group at every different phase (P＜0.01). ConclusionRat tail collagen can induce EPC involved in immigration, proliferation and pullulation in angiogenesis. The three-diamension model of EPC can be used to angiogenesis research. VEGF can mobilize and induce EPC to promote the angiogenesis.

Citation： SHI Sen,HE Yanzheng,LIU Yong,GE Hongwei,HU Bo,WEN Xuegang,ZENG Hong,ZHONG Wu,YANG Hui. Establishment and Analysis of Three-Dimensional Angiogenesis Model of Endothelial Progenitor Cell from Peripheral Blood. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2009, 16(2): 119-123. doi: Copy

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1. 1Friedrich EB, Walenta K, Scharlau J, et al. CD34-/CD133+/VEGFR-2+ endothelial progenitor cell subpopulation with potent vasoregenerative capacities[J]. Circ Res, 2006; 98(3)： e20 2Kupatt C, Horstkotte J, Vlastos GA, et al. Embryonic endothelial progenitor cells expressing a broad range of proangiogenic and remodeling factors enhance vascularization and tissue recovery in acute and chronic ischemia[J]. FASEB J, 2005; 19(11)： 1576.
2. 8Montaez E, Casaroli-Marano RP, Vilaró S, et al. Comparative study of tube assembly in three-dimensional collagen matrix and on Matrigel coats [J]. Angiogenesis, 2002; 5(3)： 167 9Moiseeva EP, Fox LH, Howells LM, et al. Indole-3- carbinol-induced death in Cancer cells involves EGFR downregnlationand is exacerbated in a 3D environment [J]. Apoptosis, 2006; 11(5)： 799.
3. Kong D, Melo LG, Gnecchi M, et al. Cytokine-induced mobilization of circulating endothelial progenitor cells enhances repair of injured arteries [J].Circulation,2004; 110(14)： 2039.
4. Enomoto S, Yoshiyama M, Omura T, et al. Microbubble destruction with ultrasound augments neovascularisation by bone marrow cell transplantation in rat hind limb ischaemia[J]. Heart, 2006; 92(4)： 515.
5. Peichev M, Naiyer AJ, Pereira D, et al． Expression of VEGFR-2 and AC133 by circulating human CD34(+) cells identifies a population of functional endothelial precursors [J]. Blood, 2000; 95(3)： 952.
6. Shmelkov SV, Jun L, St Clair R, et al.Alternative promoters regulate transcription of the gene that encodes stem cell surface protein AC133 [J]. Blood, 2004; 103(6)： 2055.
7. 潘玉先,郑振华. 人大隐静脉内皮细胞种植人工血管的实验研究 [J]. 中国普外基础与临床杂志， 2000； 7（1）： 13.
8. Wang R, Xu J, Juliette L, et al. Three-dimensional co-culture models to study prostate cancer growth， progression， and metastasis to bone [J]. Semin Cancer Biol, 2005; 15(5)： 353.
9. Desoize B, Jardillier J. Multicellular resistance: a paradigm for clinical resistance[J]. Crit Rev Oneol Hematol, 2000; 36(2-3)： 193.
10. Ferrara N， Gerber HP, LeCouter J． The biology of VEGF and its receptors [J]. Nat Med, 2003; 9(6)： 669.
11. Blacher S, Devy L, Burbridge MF, et al. Improved quantification of angiogenesis in the rat aortic ring assay [J]. Angiogenesis, 2001; 4(2)： 133.
12. Chalupowicz DG, Chowdhury ZA, Bach TL, et al. FibrinⅡ induces endothelial cell capillaary tube formation [J]. J Cell Biol, 1995; 130(1)： 207.
13. Wang HS, Hwang LL, Sue HF, et al. A simple quantitative method for evaluation of angiogenesis activity [J]. Assay Drug Dev Technol, 2004; 2(1)： 31.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Establishment and Analysis of Three-Dimensional Angiogenesis Model of Endothelial Progenitor Cell from Peripheral Blood

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