胚胎与成人视网膜神经细胞培养

目的 建立胚胎及成人视网膜神经细胞体外培养系统，为视网膜神经细胞的基础研究与药物开发提供实验模型。 方法 10～13周龄胎儿及20～40岁成人视网膜神经层用不同的酶进行消化，分散的细胞接种于预先经过包被的细胞培养盘内，加入或不加入表皮生长因子(epidermal growth factor, EGF)、成纤维细胞生长因子(fibroblast growth factor, FGF)、脑源性神经营养因子(brainderived neurotrophic factor, BDNF)、神经营养素 4(neurotrophic-4，NT-4)等培养。通过BrdU掺入、免疫组织化学及免疫荧光等方法检测培养细胞增殖并辨别细胞成分。 结果 胚胎及成人视网膜细胞可在体外连续培养100及180 d以上。加入EGF、FGF、BDNF或NT-4可明显促进胚胎与成人视网膜神经元存活，胎儿视网膜细胞增殖。与对照组相比，处理组神经元或神经节细胞的百分率较高。 结论 胚胎及成人视网膜神经细胞培养技术为视网膜神经细胞基础研究及药物开发提供了一种十分有价值的手段，加入外源性的生长因子可促进培养的视网膜神经细胞存活、增殖与分化。 (中华眼底病杂志, 2002, 18: 279-282)     

人雪旺氏细胞分泌神经营养因子浓度的测定

目的测定体外培养的人雪旺氏细胞分泌神经营养因子在各时间点的浓度并观察其时间-浓度关系。方法取人雪旺氏细胞进行体外培养,在培养后的第1天、第3天、第7天、第14天、第21天及第28天六个时间点用间接酶联免疫吸咐法测定培养液中碱性成纤维细胞生长因子(bFGF)、睫状神经营养因子(CNTF)和脑源性神经营养因子(BDNF)的浓度。结果在人雪旺氏细胞培养后的第1天、第3天、第7天、第14天、第21天及第28天时,在培养液中均可检测到bFGF、CNTF和BDNF的存在。在培养后第21天及第28天时,培养液中三种神经营养因子的浓度都显著增加。bFGF的浓度显示出高于CNTF和BDNF浓度的趋势(P<0.01)。结论培养的雪旺氏细胞能分泌神经营养因子,其浓度随培养时间的推移而升高。     

生长因子对人视网膜神经胶质细胞增殖的作用

目的：研究生长因子对培养的人视网膜神经胶质细胞增殖的作用。方法：在培养的人视网膜神经胶质细胞中分别加入表皮生长因子(epidermal growth factor, EGF,0.5～100.0ng/ml)和神经生长因子(nerve growth factor,NGF,0.05～10.0ng/ml),培养3天后用MTT法测量细胞增殖情况。结果：EGF(0.5～100.0ng/ml)和NGF(0.05～10.0ng/ml)均能刺激人视网膜神经胶质细胞的增殖,其EC₅₀(half effective concentration,半数有效浓度)分别为17.0ng/ml和 0.7ng/ml。结论：EGF和NGF均能刺激人视网膜神经胶质细胞的增殖,它们可能在增殖性玻璃体视网膜病变(proliferative vitreoretinopathy,PVR）等增殖性病变中起一定作用。 (中华眼底病杂志,1998,14:33-34) (中华眼底病杂志,1998,14:33-34)     

视神经半损伤后复合神经营养因子辅助再生的动态生物学研究

目的动态观察和探讨视神经(optic nerve,ON)1/2损伤后视网膜神经节细胞(retinea gangling cells,RGCs)的轴树突及神经元自然再生和复合神经营养因子辅助ON再生的细胞生物学变化特征,以寻找ON不全损伤后修复再生的新途径。方法①实验采用成年猫共计40只,随机分4组:正常对照组、眼球后ON1/2切断组、ON1/2切断并定时眼玻璃体内注射生理盐水对照组和ON1/2切断并定时眼玻璃体内注射脑衍化神经因子(brain-derived neurotrophic factor,BD-NF)和睫状神经营养因子(ciliary neurotrophic factor,CNTF)复合神经营养因子辅助再生组。②动物模型制作以眶外侧入路手术开眶,暴露眼球后ON,在球后5mm处准确行ON1/2切断。术后4组动物在相同视环境中饲养1、4和8个月,ON1/2切断并于玻璃体内注射BDNF和CNTF辅助再生组在术后即日注射BDNF和CNTF复合液(10ng),并在以后每周注射等量BDNF和CNTF复合液2次。在ON损伤后1、4个月和8～12个月,3次行图形视觉诱发电位(pattern reversed vi...     

脑源性神经营养因子对大鼠视网膜节细胞损伤的保护作用

目的：观察脑源性神经营养因子（brain-derived neurotrophic factor,BDNF)对Sprague-Dawly(SD)大鼠视神经损伤后视网膜节细胞（retinal ganglin cells,RGC)存活的作用。方法：将SD大鼠分为正常对照组,夹伤对照组,溶剂对照组,BDNF治疗组4组,每组20只眼。荧光金逆行标记RGC后7天,除正常组外行球后视神经夹伤,将100ng BDNF注入BDNF治疗组大鼠玻璃体腔内,分别于5,7,14,21,28天行RGC计数。结果：视神经夹伤后第7天RGC开始减少,14天时降至正常对照的70．2％,28天时降至40．5％。与正常组相比BDNF治疗组14天时RGC数开始减少,但7、14、21、28天RGC数均明显多于夹伤组（P＜0．01）。结论：在视神经夹伤后眼内注射BDNF能减少RGC的死亡,对RGC损伤有保护作用。     

荧光金逆行标记检测CNTF基因眼内转移对视神经损伤大鼠的影响

目的通过荧光金逆行标记的方法,检测腺病毒(adenovirus,Ad)介导的睫状神经营养因子(ciliary neurotrophic factor,CNTF)对视神经钳夹伤大鼠视网膜神经节细胞(retinal ganglion cell,RGC)数目的影响。方法采用钳夹视神经法制作大鼠视神经损伤模型,夹伤后向大鼠伤眼内注射Ad-CNTF,在取材前7d进行荧光金逆行标记RGC,于伤后28d,对大鼠的视网膜铺片进行荧光金标记的RGC记数。结果伤后28d,单纯损伤组、PBS组、Ad-LacZ组视网膜标记RGC数均较低;CNTF处理组视网膜标记RGC数为每视野(12.11±2.29)个(n=9),高于前3组,且差异非常显著(P<0.01)。Ad-CNTF组视网膜标记RGC数为每视野(20.93±2.67)个(n=8),在各组标记RGC数中最多,也好于CNTF组,2组间差异非常显著(P<0.01)。结论视神经损伤后眼内单次注射Ad-CNTF,可明显增加钳夹伤后28d大鼠视网膜的标记RGC数。与单次注射CNTF相比,对损伤的RGC具有更加显著的神经保护作用。     

表皮生长因子、碱性成纤维细胞生长因子对人视网膜色素上皮细胞DNA合成的影响

目的探索表皮生长因子(epidermal growth factor,EGF)、碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)对培养的人视网膜色素上皮(retinal pigment epithelium,RPE)细胞促进DNA合成的最佳刺激浓度，并对两种因子的协同作用进行探讨。方法培养的人RPE细胞第6代用于本实验。应用氚标胸腺嘧啶核苷(³H-thymidine, ³H-TdR)掺入试验及放射自显影检测EGF、bFGF对RPE细胞的促DNA合成作用。结果EGF、bFGF均可引起剂量依赖的促有丝分裂作用。在含2%血清的培养液中,EGE、bFGF作用最佳浓度为1ng/ml,明显低于无血清培养液中EGF、bFGF作用的最佳浓度(10ng/ml)。联合应用10ng/mlEGF、10ng/mlbFGF约提高RPE细胞合成DNA能力2.96倍。结论EGF、bFGF对培养的人RPE细胞具有促进DNA合成作用，且两者可产生协同效应。(中华眼底病杂志,1998,14:98-100)     

睫状神经生长因子与眼病的相关研究

睫状神经营养因子(ciliary neurotrophic factor,CNTF)是神经保护因子中被研究的最多的因子之一,细胞包囊技术(encapsulated cell technology,ECT)为CNTF的临床应用提供了安全有效的给药途径.大量的研究证据表明,CNTF对视网膜色素变性、青光眼、年龄相关性黄斑变性、视神经损伤等眼部疾病具有神经保护作用.本文就CNTF与眼病的相关研究进展进行综述.     

大鼠视网膜前体细胞的传代培养及体外分化诱导

目的了解表皮生长因子（EGF）、碱性成纤维细胞生长因子（bFGF）及睫状神经营养因子（CNTF）对其分化的影响。方法取孕18d的SD大鼠胚胎眼的视网膜，分别用悬浮法及贴壁法培养。传至第一代后转入分别含10ng／mlCNTF、20ng／mlEGF、10ng／mlbFGF、10％胎牛血清的DMEM：F12中进行诱导分化，贴壁后2、3、4、6d分别行nestin、Vimentin、Opsin及GFAP的二步法免疫组织化学染色。结果大鼠的RPCs在悬浮时呈球状生长。nestin及Vimentin染色阳性，悬浮培养传至一代，贴壁培养传至五代，传代后细胞数量不断减少并可在体外分化为含视网膜光感受器细胞及胶质细胞的混合神经元。CNTF、bFGF及EGF不同程度地增加了光感受器细胞的比例。结论RPCs可在体外培养并传代。CNTF、bFGF及EGF参与视网膜发育的调节。     

CNTF和Ad-BDNF对视神经夹伤后视网膜神经节细胞存活的影响

目的:观察大鼠视神经夹伤后玻璃体腔内注射睫状神经营养因子(CNTF)和腺病毒介导脑源性神经营养因子(Ad-BDNF)对视神经损伤后视网膜神经节细胞(RGC)存活的影响。方法:制作大鼠视神经定量夹伤模型,玻璃体腔内注射CNTF和Ad-BDNF,经上丘荧光金(FG)逆行标记RGC,计数视网膜铺片上的RGC并行统计学分析。结果:正常SD大鼠视网膜上RGC密度为2155±265个/mm2(n=12),视神经夹伤后RGC在1~2wk内下降速率最快,到3,4wk时RGC细胞数量虽仍有减少但下降速度已经明显减慢。CNTF组在视神经夹伤后1wk时视网膜RGC数显著高于对照组,但2~4wk的结果和对照组比较差异不明显。Ad-BDNF组视神经夹伤后1~4wk视网膜RGC数均显著高于对照组。结论:CNTF治疗组玻璃体腔内一次性注射CNTF可以在损伤早期2wk内为损伤的RGC提供神经营养因子,减少RGC的早期死亡。Ad-BDNF治疗组的这种保护作用可以持续到损伤后4wk,能够为RGC提供长时间地营养支持,但这种作用比较局限,可能与单一营养因子作用有关。     

Serum growth factor analysis in dry eye syndrome

Background: To perform a comprehensive serum growth factor analysis in dry eye syndrome patients and to compare this with matched controls. Methods: Six female dry eye syndrome patients and six age‐ and gender‐matched controls were recruited. Whole blood was collected, allowed to clot and then centrifuged. Serum was extracted by using sterile technique. Enzyme‐linked immunosorbent assays were performed to quantify serum growth factor levels. Results: Levels of transforming growth factor‐beta 1 and 2 (TGF‐β1 and β2), nerve growth factor (NGF), insulin‐like growth factor‐1 (IGF‐1), epidermal growth factor (EGF), acidic and basic fibroblast growth factor (FGF), keratinocyte growth factor (KGF), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), platelet‐derived growth factor‐AA, AB and BB (PDGF‐AA, AB and BB), brain‐derived neurotrophic factor (BDNF), neurotrophin‐3 (NT‐3) and glial cell line‐derived neurotrophic factor (GDNF) were quantified, and statistical analysis was performed by using the Mann–Whitney U‐test with the Bonferroni correction. Conclusions: No significant difference was found between serum growth factor levels in dry eye syndrome patients versus controls. Our study provides comprehensive analysis of serum growth factor levels in autologous serum eye drops produced from ocular surface disease patients. A knowledge of growth factor levels in serum may be important because of the increasing use of autologous serum eye drops in refractory ocular surface diseases and for an understanding of how topical serum may provide benefit.     

Presence and localization of neurotrophins and neurotrophin receptors in rat lacrimal gland

PURPOSE: To determine which of the neurotrophins (NTs)-nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT3), and neurotrophin-4/5 (NT4)-and their receptors (NTrs), TrkA, TrkB, TrkC, and p75, are present in the adult rat lacrimal gland. METHODS: RT-PCR was performed on RNA isolated from male rat lacrimal gland, using oligonucleotides specific to each NT and NTr. The presence of NT and NTr protein, was determined by Western blot analysis of lacrimal gland homogenate or membranes. The location of NTs and NTrs was determined by immunofluorescence histochemistry. Western blot analyses and immunofluorescence microscopy were performed using primary rabbit polyclonal antibodies raised against NTs and NTrs. RESULTS: RT-PCR showed positive bands at the appropriate sizes for NGF, BDNF, NT3, and NT4, and for the receptors TrkA, TrkB, TrkC, and p75. Western blot analysis confirmed these results, showing that the lacrimal gland expresses NGF, BDNF, NT3, and NT4 as well as the NTrs TrkA, TrkB, and TrkC and the p75 protein. NGF, BDNF, NT3, and NT4 were localized in the lacrimal gland acini with differing cellular distributions, whereas TrkA, TrkB, and TrkC, were localized in myoepithelial cell and ductal cell membranes. The protein p75 was expressed only on myoepithelial cell membranes. CONCLUSIONS: Members of the neurotrophin family of growth factors and their receptors are present in rat lacrimal gland, which suggests a role for NTs and their receptors in the lacrimal gland.     

大鼠视网膜神经网膜上神经营养因子的表达

目的　探讨神经生长因子 (N GF)、神经营养因子 3 (N T- 3 )及脑源性神经营养因子 (BDN F)在大鼠神经视网膜上的表达与分布。方法　 4 0 g· L- 1多聚甲醛灌注成年 Wistar大鼠 ,断头取眼球 ,放入 4 0 g· L- 1多聚甲醛 -蔗糖 (3 0 0 g· L- 1)固定液中过夜。运用免疫组化方法定性测定视网膜神经网膜上 N GF、BDN F、N T- 3的表达与分布。结果　 NG F、BDN F、NT - 3在 W istar大鼠视网膜神经网膜内均有表达 ,而且 3种神经营养因子主要分布于神经节细胞层和内核层 ,NT- 3主要分布于细胞核内 ,而N GF和 BDN F主要分布于细胞浆内。结论　 W istar大鼠视网膜神经网膜内含有丰富的神经营养因子 N GF、BDN F、N T- 3。     

Paradoxical role of BDNF: BDNF+/- retinas are protected against light damage-mediated stress

PURPOSE: Photoreceptors can be prevented from undergoing apoptosis in response to constant light by the application of exogenous neuroprotective agents, including brain-derived neurotrophic factor (BDNF). BDNF, however, cannot exert its effect directly on photoreceptors because they do not express receptors for BDNF. It has been proposed that BDNF released from Müller cells provides a feed-forward loop, increasing ciliary neurotrophic factor (CNTF) and basic fibroblast growth factor (bFGF) production in Müller cells, which may enhance photoreceptor survival. The authors hypothesized that retinas with reduced BDNF levels in which the BDNF-mediated release of neuroprotective signals is dampened are more susceptible to light-induced photoreceptor degeneration. METHODS: Young adult BDNF+/+ and BDNF+/- littermates (B6.129-BDNF(tm1-LT)) were analyzed. Retinal neurotrophin and growth factor mRNA levels were determined by quantitative RT-PCR, photoreceptor function was assessed through electroretinography, and survival was documented in morphologic sections and in TUNEL assays. Oxidative stress was assayed by measuring glutathione peroxidase activity. RESULTS: At baseline, BDNF+/- animals had significantly increased levels of glial-derived neurotrophic factor (GDNF) mRNA compared with their wild-type littermates. After light damage GDNF, CNTF, and BDNF mRNA levels dropped 14- to 16-fold in the BDNF+/+ mice but remained almost unchanged compared with baseline levels in the BDNF+/- mice. Preservation of neurotrophin levels in BDNF+/- mice correlated with photoreceptor cell survival, preservation of function, and reduced oxidative stress. CONCLUSIONS: Contrary to the hypothesis, reducing BDNF levels resulted in photoreceptor protection against light damage. Survival was paralleled by a reduction in oxidative stress and the preservation of neurotrophin levels, suggesting that chronic reduction of BDNF in the retina provides a level of preconditioning against stress.     

CNTF and BDNF have similar effects on retinal ganglion cell survival but differential effects on nitric oxide synthase expression soon after optic nerve injury

PURPOSE: To investigate the effect of ciliary neurotrophic factor (CNTF) and brain-derived neurotrophic factor (BDNF) on retinal ganglion cell (RGC) survival and nitric oxide synthase (NOS) expression in the retina during the early phase of optic nerve (ON) injury, and to examine whether intraperitoneal application of the NOS scavenger nitro-l-arginine (l-NA) could protect the injured RGCs. METHODS: RGCs were retrogradely labeled with granular blue 3 days before the ON was intraorbitally transected. RGC survival was examined 1 week after ON transection and intraocular injection of CNTF and/or BDNF, or 1 to 2 weeks after daily intraperitoneal injection of the NOS inhibitor l-NA. NOS expression was examined by NADPH-diaphorase histochemistry and neuronal NOS (nNOS) immunohistochemistry, and nNOS-positive cells were identified by various staining approaches. RESULTS: Both CNTF and BDNF significantly increased RGC survival 1 week after ON injury. In the ganglion cell layer (GCL), CNTF did not increase the number of NADPH-diaphorase positive ((+)) cells but appeared to reduce the intensity of NADPH-diaphorase staining, whereas BDNF increased the number of NADPH-diaphorase(+) cells and also appeared to enhance the intensity of NADPH-diaphorase staining. In the GCL, amacrine cells but not RGCs were nNOS(+). Some macrophages were also nNOS(+). In contrast, no amacrine cells were nNOS(+) in the inner nuclear layer. Daily intraperitoneal injection of l-NA at appropriate concentrations promoted RGC survival for 1 or 2 weeks after ON injury. CONCLUSIONS: Both CNTF and BDNF protected RGCs after ON injury. CNTF and BDNF acted differently on NOS expression in the GCL. Intraperitoneal injections of l-NA at appropriate dosages enhance RGC survival.     

Rod Outer Segment Maintenance Is Enhanced in the Presence of bFGF, CNTF and GDNF

We employed a morphological assay of outer segment collapse to determine if growth factors or other supplements directly affect dissociated rod photoreceptors in vitro. The morphological changes in outer segments were correlated with the light responsiveness of rods. Time-lapse video microscopy was used to observe the collapse of rod outer segments from isolated single cells and small clumps of cells. A consistent pattern of outer segment collapse into the inner segment was observed, yielding a convenient assay of the effects of neurotrophic factors on photoreceptor functional maintenance. The functional state of rods, defined as light-responsiveness, was measured with suction electrode recordings and matched with the various stages of outer segment collapse. Ciliary neurotrophic factor (CNTF) and glial cell-line-derived neurotrophic factor (GDNF) at a high concentration, yielded statistically significant improvements in rat outer segment survival times. Basic fibroblast growth factor (bFGF), which rescues photoreceptors in several rodent models of retinal degeneration, produced a significant increase in survival time in the presence of the cofactor heparin. In 4 out of 10 cases using human tisue, bFGF also yielded a significant increase in survival times. When brain-derived neurotrophic factor (BDNF) was applied to rat rods, outer segment survival times did not change. Outer segments collapsed more quickly when either pigment epithelial cell derived factor (PEDF) or sugarN-acetyld-galactosamine (NAD-gal) were present. Our results show that rod photoreceptors can respond to bFGF, GDNF and CNTF in vitro and provide evidence for a direct effect of these neurotrophic factors on rods. The rapid collapse of isolated photoreceptors in this model provides a convenient means for testing various neurotrophic agents and the induced cellular responses.     

睫状神经营养因子对培养大鼠视网膜神经节细胞的影响

目的 观察不同浓度睫状神经营养因子(ciliary neurotrophic factor，CNTF)对培养大鼠视网膜神经节细胞（retinal ganglion cell，RGC）生长、存活的影响。 方法 取15只生后2～3d Wistar大鼠视网膜组织进行细胞培养，通过Thy-1单克隆抗体免疫细胞化学对培养的RGC进行鉴定。实验分对照组和10、20、40 ng/mlCNTF组（Ⅰ、Ⅱ、Ⅲ组），记录RGC存活时间，将培养第3、5、7天的RGC行四甲基偶氮唑盐（methylthio tetrazole,MTT）法测量吸光度(A)值［旧称光密度(OD)］。 结果 Thy-1单克隆抗体免疫组织化学检查显示培养3d的存活细胞90%以上为RGC。细胞存活期间实验组与对照组细胞均无明显突起,细胞体积无明显增大，实验组细胞存活时间比对照组长3~4d。培养第5、7d，Ⅰ组A值分别为0.075 8±0.0139、0.0693±0.0113，Ⅱ组A值分别为0.0902±0.0114、0.0825±0.0125，Ⅲ组A值分别为0.0792±0.0133、0.0653±0.0086，对照组A值分别为0.0620±0.0071、0.0513±0.0068。实验组与同时间对照组A值相比差异有显著性的意义（Ⅱ组与对照组相比P＜0.01，Ⅰ、Ⅲ组与对照组相比P＜0.05）。 结论 一定浓度的CNTF能促进培养大鼠RGC的存活，对RGC形态无影响。 (中华眼底病杂志, 2002, 18: 283-285)     

Characterization of genetically modified human retinal pigment epithelial cells developed for in vitro and transplantation studies

PURPOSE: To develop, by specific genetic modification, a differentiated human retinal pigment epithelial (RPE) cell line with an extended life span that can be used for investigating their function in vitro and for in vivo transplantation studies. METHODS: Primary human RPE cells were genetically modified by transfecting with a plasmid encoding the simian virus (SV)40 large T antigen. After characterization, two cell lines, designated h1RPE-7 and h1RPE-116, were chosen for further investigation, along with the spontaneously derived RPE cell line ARPE-19. Factors reported to be important in RPE and photoreceptor cell function and survival in vivo were examined. RESULTS: Both h1RPE-7 and h1RPE-116 cells exhibited epithelial morphology, expressed cytokeratins, and displayed junctional distribution of ZO-1, p100-p120 and beta-catenin. The cells expressed mRNA for RPE65 and cellular retinaldehyde-binding protein (CRALBP) and the trophic and growth factors brain-derived neurotropic factor (BDNF), ciliary neurotrophic factor (CNTF), basic fibroblast growth factor (bFGF), pigment epithelium-derived factor (PEDF), nerve growth factor (NGF), platelet-derived growth factor (PDGF)-alpha, insulin-like growth factor (IGF)-1, and vascular endothelial growth factor (VEGF). Secreted BDNF, bFGF, and VEGF, but not CNTF, were identified in cell supernatants. The cell lines constitutively expressed HLA-ABC, CD54, CD58, and CD59. After activation with IFN-gamma both HLA-ABC and CD54 were upregulated, and the expression of HLA-DR was induced. Both cell lines failed to express CD80, CD86, CD40, or CD48 in vitro and in a mixed lymphocyte reaction were unable to induce T-cell proliferation. Fas ligand (CD95L) was not detected in vitro by RT-PCR. Similar results were obtained with the ARPE-19 cell line. CONCLUSIONS: RPE lines h1RPE-7 and h1RPE-116 retain many of the morphologic and biochemical characteristics of RPE cells in vivo and may serve as a source of cells for in vitro analysis of RPE cell function, as well as for orthotopic transplantation studies.     

Effect of growth factors on bovine retinal pigment epithelial cell migration and proliferation

BACKGROUND: To examine the effects of platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), acidic fibroblast growth factor (aFGF), insulin-like growth factor-1 (IGF-1), epidermal growth factor (EGF), and transforming growth factor beta 2 (TGF(beta2)) on bovine retinal pigment epithelial (RPE) cell migration and proliferation. MATERIALS AND METHODS: Cultured bovine RPE cells were treated with 10 ng/ml PDGF, bFGF, aFGF, IGF-1, EGF, or TGF(beta2). RPE cell migration studies were performed in multiwell plates confluently covered with RPE cells. After inhibition of proliferation and denudation of half of each well, cells were incubated with various growth factors. Migration was measured as the number of cells that had entered the denuded area after 20 h. RPE cell proliferation was determined by [(3)H]thymidine incorporation after growth factor stimulation for 24 h. RESULTS: RPE cell migration was significantly enhanced after incubation with PDGF (stimulation of 213% compared to the negative control, p = 0.002), bFGF (206%, p = 0.003), aFGF (175%, p = 0.003), IGF-1 (150%, p = 0.003), and EGF (144%, p = 0.003). RPE cell proliferation was stimulated by bFGF (322% compared to the negative control, p < 0.005), PDGF (119%, p < 0.005), aFGF (121%, p < 0.005), and EGF (94%, p < 0.005). IGF-1 showed no significant effect on RPE cell proliferation; TGF(beta2) displayed no effect on RPE cell migration nor on proliferation. CONCLUSIONS: The peptide growth factors PDGF, bFGF, aFGF, IGF-1, and EGF play an important role in initiating RPE cell migration. Basic FGF, PDGF, aFGF, and EGF stimulate RPE cell DNA synthesis.