《About the role of platelet-derived growth factor on human retinal pigment epithelial cells（血小板源生长因子的作用对人类视网膜色素上皮细胞）》由会员分享，可在线阅读，更多相关《About the role of platelet-derived growth factor on human retinal pigment epithelial cells（血小板源生长因子的作用对人类视网膜色素上皮细胞）（14页珍藏版）》请在金锄头文库上搜索。
1、1About the role of platelet-derived growth factor on human retinal pigment epithelial cellsAuthor: Secretary the Walkers benefits sickness Hanquan Hong Du Hongjun Keywords growth factor Keywords: growth factor, platelet-derived; human retinal pigment epithelial cells; proliferation; transitional Abs
2、tract: Objective To observe the platelet-derived growth factor (PDGF on human retinal pigment epithelial cells (hRPE proliferation and migration ability. Vitro 4 to 6 human retinal pigment epithelial cells into DMEM group (modified Eagle medium liquid containing 20mL L-1 calf serum DMEM group (20g L
3、-1 DMEM, each group with different concentrations (0.1,1,5,10,50 and 100g L-1 of PDGF, and then by MTT observed colorimetric analysis of its role in the proliferation of hRPE; application of the the hRPE injury model counting cells into the defect area, quantitative observation of PDGF on the migrat
4、ion of hRPE Results proliferative effect: of 0.1 100g L-1 of PDGF can promote promoting proliferation proliferation of hRPE of cells, DMEM group 10g L-1, 2the most obvious, the proliferation rate of 153%; 20g L-1 the DMEM group of 50 100g L-1 proliferation was 174%, the strongest effect. transitiona
5、l role: PDGF significantly promote hRPE transitional, and a dose-dependent manner, 50 100g L-1 to promote transitional strongest DMEM group migration rate was 510%, 20 mL L-1 FBS + DMEM group of the 640 % Conclusion PDGF can promote the proliferation and migration of hRPE have serum can strengthen t
6、hat role, suggesting that it may play an important role in the pathogenesis of PVR. Keywords: platelet-derived growth factor; human retinal pig-ment epithelium cells; proliferation; migration Abstract: AIM To investigate the effects of platelet-derived growth factor on the proliferation and migratio
7、n of cultured human retinal pigment epithelium cells (hRPE.METHODS Cultured hRPE of the4 6th passage were devided into two groups DMEM (Delbeccos modified Eagles mediumand20mL L- 1 FCS + DMEM (20g L-1 DMEM). PDGF (0.1 100g L-1 was added to medium.RPE prolifera-tion was mesured by colorimetric 3assay
8、 for cellular growth and survival (MTT assayand Migration was assessed by us- ing an in vitro wound healing model in which a small area of a confluent monolayer of hRPE was denuded with a razor blade, the cultures were subsequently incubated with medi-um, migration was measured after24hours as the n
9、umber of cells that had entered the denuded area . RESULTS Pro-liferation: PDGF stimulated the proliferation of hRPE.The maxium proliferation rate of RPE was153% at10g L-1 in DMEM and174% at50 100g L-1 in20g L-1 DMEM. Migration: PDGF also stimulated hRPE migration in a dose-dependent manner.At50 100
10、g L-1, the maxium migration rate were510% and640% in DMEM and20g L-1 DMEM.CONCLUSION PDGF stimulates hRPE prolif-eration and migration, the presence of serum can increase this effects, which imply that PDGF may play an important role in the development of proliferative vitreoretinopathy. 0 Introduct
11、ion Vitreous and subretinal fluid in proliferative vitreoretinopathy patients detected significantly higher 4platelet-derived growth factor (PDGF 1, a number of in vitro experiments showed that, in cultured retinal pigment epithelial cells (hRPE autocrine or paracrine PDGF and expression of its rece
12、ptor 2 due to the factor of a variety of cells and promote mitotic activity and chemotaxis, and is involved in the whole process of wound healing 3, we use in vitro method to observe PDGF on human retinal pigment epithelial cells (hRPE proliferation and migration ability. 1 Materials and methods 1.1
13、 Materials Delbeccos modified Eagles medium (DMEM, Gibco, USA, fetal calf serum (Hangzhou Evergreen, trypsin (Gibco, USA ,3-2, 5 - diphenyltetrazolium triazole bromine salt (MTT, Sigma, USA, people Recombinant PDGF-BB (Sigma, USA, 12 hole, 96-well culture plate (NUNC, Denmark, mitomycin C (Sigma, US
14、A, dimethyl sulfoxide (DMSO, Sigma, USA), Bio-Red550-type enzyme-linked immunosorbent detector (Japan. 1.2 Methods of human RPE cells donated by the Xijing Hospital Associate Professor Hui Wang Yusheng 4 four different sources of human retinal pigment epithelial 5cells, 4 to 6 used in the experiment
15、. Recovery and culture methods with Wang Lin et al 5 report. Proliferation assay 100mL L-1 calf serum-containing DMEM and the cell concentration adjusted to 2 107 L-1, seeded in 96-well plates (200 L / well, 37 C overnight in 50mL L-1 CO2 the hatched box, suck abandoned old medium, divided into two
16、groups: DMEM and DMEM (20 g L-1 DMEM containing 20mL L-1 calf serum in each group, respectively, with different concentrations of PDGF (0,0.1,1,10,50 and 100g L-1, each concentration of six wells, and cultured for 72h, MTT (20L / hole 5g L-1, cultured for 4h, the culture medium was aspirated off and washed twice with PBS. added DMSO (150L / well, and shaken for 10min, 4
