首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到18条相似文献,搜索用时 109 毫秒
1.
目的:探索青光眼病人小梁细胞体外培养和保存方法。方法:从小梁切除术取得的巩膜内板层,应用组织培养方法进行病人小梁细胞体外培养,完成鉴定工作。并将第四代的青光眼病人小梁细胞冻存,冻存2周,1个月,2个月,半年后,将细胞复苏,观察复苏后小梁细胞的生长情况。结果:青光眼病人小梁组织培养出的细胞经鉴定为小梁细胞,培养的小梁细胞经冷冻保存,复苏率超过80%。结论:青光眼病人小梁细胞体外培养及冻存复苏成功,使  相似文献   

2.
背景原发性开角型青光眼(POAG)是一种常见致盲性眼病,其特点是房水外流阻力增加导致眼压增高。位于房水外流通道的小梁网调节房水的外流,因此研究小梁网细胞的生物学特性有着重要的意义。目的探讨POAG小梁细胞体外培养的方法及其生物学特性。方法经小梁切除术收集8例开角型青光眼患者患眼的带小梁网的深层巩膜组织块进行体外原代和传代培养,用鼠抗人层黏连蛋白(LM)单克隆抗体、兔抗人纤维连结蛋白(FN)单克隆抗体、鼠抗人神经元特异性烯醇化酶(NSE)单克隆抗体进行免疫组织化学检测以对传代细胞进行鉴定,在透射电子显微镜下对传代细胞的超微结构进行观察,并将传代小梁细胞的生物学特性与本研究组前期培养的正常小梁细胞进行比较。结果组织块培养10d左右,可见细胞从其边缘向外生长。传代细胞在4d内处于对数生长期,其后进入平台期,第7天细胞基本融合。第3代POAG小梁细胞及正常人眼小梁细胞中可见FN、LM和NSE均呈阳性表达,证实传代细胞为小梁细胞,而空白对照组细胞未见FN、LM和NSE表达。第3代POAG小梁细胞和正常小梁细胞中FN的A450值分别为0.354±0.06和0.26±0.01,LM的A450值分别为0.34±0.03和0.25±0.02,差异均有统计学意义(FN:t=14.446,P=0.001;LM:t=9.346,P=0.001)。与正常小梁细胞比较,第3代POAG小梁细胞表面的微绒毛、细胞质的溶酶体及吞噬小泡含量减少。结论采用组织块培养法可成功在体外培养POAG小梁细胞,该研究结果为研究青光眼的发病机制提供了细胞学基础。  相似文献   

3.
目的建立体外培养的人小梁(human trabecular meshwork,HTM)细胞,并观察小梁细胞细胞骨架与细胞连接的形态学特征。方法体外培养的HTM细胞传代后,免疫组化方法鉴定,电镜观察小梁细胞的超微结构,激光共聚焦显微镜观察actin、vinculin、β-catenin的表达。结果通过光镜、透射电镜、扫描电镜和免疫组化的方法证实体外培养的细胞为人眼小梁细胞。人眼小梁细胞的细胞骨架与细胞连接丰富。结论体外培养的小梁细胞细胞骨架结构明显,证明该细胞有收缩能力;β-连接素与纽蛋白着染明显,说明体外培养的小梁细胞,其细胞与细胞间、细胞与细胞基质间的连接仍具有活体小梁细胞的特性。  相似文献   

4.
利用体外培养方法,研究了牛小梁细胞对鸡红细胞的吞噬过程及所致的细胞损伤.结果发现小梁细胞在体外对鸡红细胞有较强的吞噬能力,完整的鸡红细胞被吞入细胞内需8小时,但随着吞噬过程的延长小梁细胞的形态和结构发生损伤性改变.因此,吞噬过程所致的小梁细胞损伤或细胞数减少,可能与某些继发性青光眼的发生或老年性青光眼时的小梁细胞减少有关.  相似文献   

5.
目的 探讨国人蒙古族原发性闭角型青光眼的发病机制,为治疗蒙古族原发性闭角型青光眼提供临床依据.方法 采用病例对照研究方法.研究分为蒙古族正常人组,蒙古族青光眼组,汉族正常人组,汉族青光眼组.应用亚硝酸还原酶法测房水及血清一氧化氮浓度;应用流式细胞仪测定小梁细胞的凋亡率;利用透射电镜观察小梁组织的超微结构;利用UBM观察测量眼前段生物学指标;并对四组数据进行统计学处理.结果 四组房水及血清一氧化氮浓度性差异无统计学意义.蒙古族和汉族眼小梁细胞凋亡率比较,有种族差异,小梁网的超微结构亦有差异.蒙古族与汉族比较正常人眼前段结构有角膜小、前房浅、晶状体厚且相对位置偏前,统计学处理有显著差异.结论 蒙古族闭角型青光眼的高发病率与蒙古族人眼前段的结构异常、小梁细胞凋亡率高、小梁网的超微结构异常等因素有关,其中眼前段的结构异常是发病的高危因素.  相似文献   

6.
人眼小梁细胞体外培养,冻存与复苏   总被引:2,自引:0,他引:2  
葛坚  卓业鸿 《眼科学报》1998,14(2):73-75
目的:建立人小梁细胞体外培养及对其进行冻存、复苏研究。方法:应用组织块培养方法进行人小梁细胞体外培养,将第3代的小梁细胞冻存,冻存1周、2周、1月、2月后细胞给予复苏,观察复苏后细胞的生长情况。结果:人眼小梁细胞体外培养成功。冻存的小梁细胞复苏成功,所有复苏率超过90%。结论:人眼小梁细胞体外培养及冻存复苏成功,为以后构建小梁细胞的cDNA文库,为筛选青光眼发病的相关基因提供有利的实验基础。眼科学报1998;14:73—75。  相似文献   

7.
人眼小梁细胞体外培养及传代实验   总被引:1,自引:1,他引:0  
Dai W  Li M 《中华眼科杂志》1998,34(2):121-123
目的探索体外培养人眼小梁细胞的方法,为进一步研究原发性开角型青光眼的发病机理提供实验的依据。方法用器官培养及组织块培养两种方法,进行人眼小梁细胞的原代培养和传代实验,并同时对照培养人眼巩膜成纤维细胞,对二者从形态学上进行比较。结果(1)器官培养较单纯组织块培养更易获得原代小梁细胞;(2)传3~5代人眼小梁细胞处于最稳定阶段,可利用此阶段细胞进行多种体外实验。结论体外培养人眼小梁细胞可获得生长形态及特征稳定的小梁细胞。  相似文献   

8.
9.
原发性开角型青光眼(POAG)是青光眼主要致盲性眼病之一,其发病机制尚不清楚。近年来许多研究已表明,POAG眼压升高是由于房水排出道的病变,使房水排出阻力增加所致。阻力的部位主要在小梁网,特别是近管组织。阻力的形成与衬覆在小梁柱表面的小梁细胞形态与功能有关。由于直接研究体内小梁细胞非常困难,国外许多学者用小梁细胞体外培养的方法,研究小梁细胞的形态结构与功能活动,发现POAG的发生与小梁细胞的异常密切相关,小梁细胞体外培养是探索POAG病因和发病机制的有效途径。  相似文献   

10.
目的选择性行牛眼上皮样小梁细胞的体外培养,为进一步研究原发性开角型青光眼提供实验材料。方法利用组织块法进行小梁细胞的原代培养,之后利用机械划除法、酶消化法、反复贴壁法,对牛眼上皮样小梁细胞进行选择性培养和传代。结果成功选择性培养出形态、性状良好的牛眼上皮样小梁细胞。结论选择性牛眼上皮样小梁细胞的体外培养所获细胞形态单一,可以用此种方法进行牛小梁细胞的体外培养并为实验应用做好准备。  相似文献   

11.
Jian  Ge  Minkai  Lin 《眼科学报》1998,14(3):134-137
Purpose : To establish the culture system of human glaucomatous trabecular cells in vitro and study their ultrastructures.Methods : The trabecular specimens from trabeculectomy were cultured in vitro and passaged 3 times, then identified. Moreover, the glaucomatous cells were observed with electron microscope while compared with the normal ones.Result: Cultured human glaucomatous trabecular cells were obtained. The ultrastructure of the cells showed the decrease in vilious project, coated vesicle and lysosomal inclusion. Conclusion : The establishment of human glaucomatous trabecular cells culture in vitro made the culture system more perfect. The morphologic changes might be related to the abnormal functions of human trabecular meshwork cells. Eye Science 1998; 14 : 134 - 737.  相似文献   

12.
Yehong  Zhuo  Jian  Ge 《眼科学报》1999,15(1):46-50
Background: To study the glucocorticoid receptor (GR) and the associated gene regulation in the pathogenesis of glucocorticoid- induced glaucoma (GIG) in Chinese patients.Methods: The trabecular cells of normal individuals and patients with GIG were cultured in vitro. By using polymerase chain reaction (PCR),gene fragments on GR DNA binding sites of trabecular cells were amplified. The product was detected by gel electrophoresis.Results: The trabecular cells were cultured successfully in normal individuals and patients with GIG in vitro. A single PCR product was obtained in both two groups with the same size of 545 base pairs.Conclusion: There is not any difference in gene on the GR DNA binding sites between normal individuals and patients with GIG. The results suggest the difference in mRNA or other functional genes. Eye Science 1999 ; 15 ; 46 - 50.  相似文献   

13.
PURPOSE: Depletion of trabecular meshwork cell numbers is a feature of the outflow system in aging and in primary open-angle glaucoma. It is possible that migration stimulated by factors present in aqueous humor may contribute to the cell loss. This investigation assessed the chemoattractant potential of glaucomatous and nonglaucomatous human aqueous humor and fibronectin, one of its constituents, on a range of cultured trabecular meshwork cell lines. METHODS: Migration was assessed in 48-well modified Boyden chambers. The potential migratory stimulants were soluble fibronectin and glaucomatous and nonglaucomatous aqueous humor. The glaucomatous aqueous samples were collected from patients undergoing trabeculotomy for primary open-angle glaucoma and the normal aqueous from normal bovine eyes and patients undergoing cataract surgery. The target cell types were normal human and bovine meshwork cells grown from explants and two human transformed meshwork cell lines from a normal (HTM-5) and a glaucomatous (HTM-3) source. RESULTS: Soluble fibronectin stimulated all the target cells to migrate with an optimal concentration ranging from 1 to 30 microg/ml, and Zigmond Hirsch checkerboard analysis indicated that both chemotaxis and chemokinesis took place. All the aqueous humor samples stimulated migration of the meshwork cell lines at an optimal concentration of 200 microl/ml. Glaucomatous aqueous humor stimulated a greater migratory response than nonglaucomatous aqueous for two of the four target cell types (P < or = 0.03). Neutralization of the fibronectin content of nonglaucomatous and glaucomatous aqueous by addition of excess anti-fibronectin antibody indicated that fibronectin could account for 35% to 80% of the migratory activity of the aqueous. CONCLUSIONS: Aqueous humor contains potentially powerful chemoattractants for trabecular meshwork cells. The activity of one of these constituents, fibronectin, has been accounted for by this study. Glaucomatous aqueous appears to be as good and in some cases a better migratory stimulant than nonglaucomatous aqueous in vitro. The migratory evidence points to a trend that may help to explain cell loss in the aging meshwork and possibly some of the extra loss in primary open-angle glaucoma.  相似文献   

14.
Fibronectin in human trabecular drainage channels   总被引:2,自引:0,他引:2  
Fibronectin, an extracellular glycoprotein, has been shown to be produced by human trabecular cells in culture by our group as well as Polansky and co-investigators. Studies of Rodrigues et al suggested that fibronectin may be one of several glycoproteins found in increased amounts in the corneoscleral trabecular meshwork of glaucomatous eyes. The authors have developed a sensitive immunoassay utilizing avidin-biotinylated enzyme complex (ABC) to detect low levels of fibronectin in frozen sections of human eyes. The authors have used this immunoassay together with a perfusion technique to demonstrate distribution patterns of fibronectin present in human aqueous drainage channels. The authors found that fibronectin is present in larger quantities in the aqueous drainage channels than in the surrounding tissues in 18 eyes from older patients.  相似文献   

15.
人眼小梁细胞体外培养及生物学特性研究   总被引:5,自引:1,他引:4  
目的:建立人眼小梁细胞培养方法并研究其生物学特性。方法:以体外组织块培养的方法获得培养的人小梁细胞,应用光镜、电镜观察细胞的形态学特征,并观察其免疫组化特性和细胞的生长曲线。结果:光镜下小梁细胞为扁平多角形、单层生长;电镜下细胞连接为点粘连和缝隙连接、细胞表面可见微绒毛、胞浆细胞器丰富;免疫组化染色对抗纤维连接蛋白(Anti—FN)、抗层粘连蛋白(Anti—LN)、抗神经元特异性烯醇化酶(Anti—NSE)单抗呈阳性,对抗第Ⅷ因子(Anti-ⅧFactor)单抗呈阴性;传代小梁细胞繁殖时间较长,10天后为平台期。结论:根据体外培养的小梁细胞的形态特点、生长特征、免疫组化特性可对其进行鉴定。人小梁细胞体外培养的成功,为在细胞和分子水平研究青光眼的发病机制提供了有利的条件。眼科学报 1996;12:64—69。  相似文献   

16.
PURPOSE: Isolation and culture of human trabecular meshwork (TM) cells from primary open-angle glaucomatous (POAG) tissue has proven difficult. The objective of this study was to directly compare the utility of two different isolation methods to obtain viable human TM cells from POAG whole eye tissue. METHODS: Using a blunt dissection technique, human TM tissue was obtained from four pairs of donor eyes (67, 77, 81 and 82 years) with a documented history of POAG. TM tissue from one eye was explanted into tissue culture. TM from the contralateral eye was digested with a collagenase mixture and seeded onto culture plates. RESULTS: Primary cell isolates were obtained from all donors with both techniques. However, only cells obtained using the digestion method (3 of 4 TMs) could be passaged for expansion and freeze-downs (3 x 107 second passage cells/donor). None of the cells obtained from explanted TMs could be passaged. Cells from successful isolations were of uniform size, possessed typical TM morphology and had doubling times < 48 hours. CONCLUSION: These results demonstrate a clear advantage to digesting the extracellular matrix of glaucomatous TM tissue to obtain sufficient numbers of healthy cells for use in experiments. In contrast to cells obtained from explants, cells liberated from POAG TM tissue by digestion appear indistinguishable morphologically and behaviorally from "normal" TM cells.  相似文献   

17.
Glucocorticoid treatment in vivo can produce a glaucoma similar in many ways to POAG. Treatment of trabecular meshwork cells in culture with dexamethasone allows the study of biochemical aspects of this disease process. The effects of dexamethasone on the expression of integrins and laminin in both normal and glaucomatous cultured human trabecular meshwork cells were evaluated. Human trabecular meshwork cell lines were cultured for 18 days in the presence or absence of 10−7mdexamethasone. Radioimmunoprecipitation was used to determine the relative expression of five αintegrin subunits. Laminin expression was evaluated with Western blots. Laminin was increased in all cell lines following dexamethasone treatment. α2, α5 and αV integrin chains showed consistent dexamethasone-induced changes in expression, while α3 and α4 subunits did not. There were no differences in the expression patterns for any of these integrin subunits between normal and glaucomatous cell lines. Increased laminin deposition as seen in this study with dexamethasone treatment may be partially responsible for the decreased outflow facility seen in both steroid-induced glaucoma and in POAG.  相似文献   

18.
PURPOSE: We previously reported a novel cytoskeletal protein with a myosin-like domain which is localized in the ciliary rootlet and basal body of connecting cilium of photoreceptor and hence we named it 'myocilin'. It was soon realized that myocilin is identical to a protein called TIGR (trabecular meshwork inducible glucocorticoid response protein) which was found to be responsible for the pathogenesis of juvenile open angle glaucoma. In this study, we employed in situ RNA hybridization to examine the myocilin (MYOC)/ TIGR gene expression in the trabecular meshworks of glaucomatous and nonglaucomatous eyes. METHODS: The glaucomatous specimens were obtained by trabeculectomy from the patients with primary open angle glaucoma (POAG), chronic angle closure glaucoma (CACG) and steroid glaucoma, respectively, and the nonglaucomatous specimens were obtained from a victim of traffic accident at autopsy and from a patient with maxillary sinus carcinoma at enucleation for the operation. The in situ RNA hybridization was carried out with digoxigenin-labeled sense and antisense RNA probes. RESULTS: In all cases, hybridization signals were detected primarily in the trabecular meshwork cells and secondarily in the fibroblast-like cells of corneoscleral wall. CONCLUSIONS: Myocilin gene is expressed clearly in the trabecular meshwork cells of both glaucomatous and nonglaucomatous eyes.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号