Isolation and culture of iris pigment epithelium from iridectomy specimens of eyes with and without exfoliation syndrome.
Isolation and culture of iris pigment epithelium from iridectomy specimens of eyes with and without exfoliation syndrome.
复制标题
从患有和不患有剥脱综合征的眼睛虹膜切除标本中分离和培养虹膜色素上皮。
DOI：
10.1001/archopht.1997.01100150091015
复制
发表时间：
1997
影响因子：
--
通讯作者：
R. Ritch
中科院分区：
文献类型：
--
作者：
Dan;Steven A. McCormick;R. Ritch
目标
从患有或不患有剥脱综合征的眼睛虹膜切除手术标本中培养虹膜色素上皮 (IPE)。
方法
处理IPE以获得单细胞悬液。细胞在 Ham F12 营养混合物中培养，其中补充有 30% 胎牛血清、50 微克/mL [校正]庆大霉素和 2 mmol/L 谷氨酰胺。汇合后，使用0.125%胰蛋白酶-0.01%依地酸溶液分离细胞，重悬、稀释并传代培养。通过透射电子显微镜研究了原代培养物和继代培养物的 IPE。进行免疫细胞化学染色。
结果
在剥脱综合征患者的IPE原代培养物中，在细胞表面发现了弯曲的、交叉带状的细原纤维（直径，10-15 nm；周期性，10-14 nm）。在细原纤维的外部发现了较粗的原纤维（直径，24-48 nm；周期性，24-36 nm）。亚培养物主要含有细原纤维。 IPE 细胞用抗细胞角蛋白、S100 蛋白和波形蛋白抗体染色呈阳性。
结论
可以从患有剥脱综合征的眼睛中成功培养虹膜色素上皮。研究体外剥脱材料的产生应提供有关剥脱综合征发病机制和剥脱材料性质的信息。从手术标本中培养正常IPE为IPE体外生长调节和药理学研究提供了来源。
OBJECTIVE
To culture iris pigment epithelium (IPE) from surgical iridectomy specimens of eyes with and without exfoliation syndrome.
METHODS
The IPE was treated to obtain a single cell suspension. Cells were cultured in Ham F12 nutrient mixture, which was supplemented with 30% fetal bovine serum, 50-micrograms/mL [corrected] gentamicin, and 2-mmol/L glutamine. After confluence, the cells were detached using a 0.125% trypsin-0.01% edetic acid solution, resuspended, diluted, and subcultured. The IPE from primary cultures and subcultures was studied by transmission electron microscopy. Immunocytochemical staining was performed.
RESULTS
In the primary cultures of IPE from patients with exfoliation syndrome, curved, cross-banded, fine fibrils (diameter, 10-15 nm; periodicity, 10-14 nm) were found on the cell surface. Thicker fibrils (diameter, 24-48 nm; periodicity, 24-36 nm) were found external to the fine fibrils. Subcultures contained mainly fine fibrils. The IPE cells stained positively with anticytokeratin, S100 protein, and vimentin antibodies.
CONCLUSION
Iris pigment epithelium can be successfully cultured from eyes with exfoliation syndrome. Studying the production of exfoliation material in vitro should provide information about the pathogenesis of exfoliation syndrome and about the nature of the exfoliation material. The cultivation of normal IPE from surgical specimens provides a source for the study of the growth regulation and pharmacophysiology of IPE in vitro.
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假性剥脱材料含有弹性微原纤维相关糖蛋白。
DOI：
--
发表时间：
1986
期刊：
Transactions of the American Ophthalmological Society
影响因子：
--
作者：
Streeten,BW;Gibson,SA;Dark,AJ
通讯作者：
Dark,AJ
结膜假性剥脱性纤维病。
影响因子：
13.7
作者：
Streeten,BW;Bookman,L;Ritch,R;Prince,AM;Dark,AJ
通讯作者：
Dark,AJ
影响因子：
4.4
作者：
Freddo,TF
通讯作者：
Freddo,TF
影响因子：
4.2
作者：
STREETEN, BW;DARK, AJ;HOEPNER, JA
通讯作者：
HOEPNER, JA
假性剥脱综合征的临床前诊断。
DOI：
10.1001/archopht.1987.01060080078032
发表时间：
1987
期刊：
Archives of ophthalmology (Chicago, Ill. : 1960)
影响因子：
--
作者：
Prince,AM;Streeten,BW;Ritch,R;Dark,AJ;Sperling,M
通讯作者：
Sperling,M