Adult human retinal cells in culture. Identification of cell types and expression of differentiated properties.
Adult human retinal cells in culture. Identification of cell types and expression of differentiated properties.
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培养中的成人视网膜细胞。
DOI：
10.1016/s0014-4827(85)80043-4
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发表时间：
1985
影响因子：
3.7
通讯作者：
Bridges,CD
中科院分区：
文献类型：
--
作者：
Oka,MS;Frederick,JM;Landers,RA;Bridges,CD
描述了一种在补充有神经生长因子（NGF）的无血清N2培养基中培养成年哺乳动物视网膜神经元的方法。在分散的人视网膜培养物中，神经元的鉴定是基于形态学、结合破伤风毒素的免疫细胞化学定位以及通过高亲和力摄取机制积累的3 H-神经递质候选物（γ-氨基丁酸、甘氨酸、多巴胺）的放射自显影定位。神经元不会附着在玻璃或塑料基底上，因此本研究使用接种在饲养层上的神经元进行。附着的初始阶段需要血清。饲养层来源于在血清存在下接种在玻璃或塑料上并随后传代的视网膜细胞。由于这些细胞表现出胶质细胞酸性蛋白（GFAP）的免疫反应性，他们被初步确定为胶质细胞的起源。在这些条件下，神经元和胶质细胞特异性特性可保留长达28天。用兔抗牛间质视黄醇结合蛋白（IRBP）抗体，通过免疫印迹技术证明了饲养层神经细胞培养液中存在IRBP。在单独培养饲养层的培养基中未检测到IRBP。IRBP的生物合成通过培养物与[35 S]甲硫氨酸的孵育来证明。免疫沉淀[35 S]IRBP仅在含有神经元的培养基中检测到;饲养层细胞不合成和分泌这种糖蛋白。这些发现与IRBP（interphotoreceptor matrix的135 K成分）在体内由神经元细胞（特别是光感受器）合成的假设一致。
A method for culturing adult mammalian retinal neurons in serum-free N2 medium supplemented with nerve growth factor (NGF) is described. Identification of neurons in cultures of dispersed human retina was based upon morphology, immunocytochemical localization of bound tetanus toxin, and autoradiographic localization of3H-neurotransmitter candidates (γ-aminobutyric acid, glycine, dopamine) accumulated by high-affinity uptake mechanisms. Neurons would not attach to glass or plastic substrates, consequently the present studies were performed using neurons plated upon a feeder layer. Serum was required for the initial phase of attachment. The feeder layer was derived from retinal cells that had been plated on glass or plastic in the presence of serum and had later been passaged. Since these cells exhibited glial fibrillary acidic protein (GFAP) immunoreactivity, they were tentatively identified as being glial in origin. Under these conditions, neuron- and glia-specific properties were retained up to 28 days. The presence of interstitial retinol-binding protein (IRBP) in medium of cultures of neuronal cells on feeder layers was demonstrated by an immunoblot technique using rabbit antibovine IRBP antibodies. No IRBP was detected in medium in which the feeder layers alone had been cultured. IRBP biosynthesis was demonstrated by incubation of the cultures with [35S]methionine. Immunoprecipitable [35S]IRBP was detected only in medium from cultures containing neurons; cells of the feeder layer did not synthesize and secrete this glycoprotein. These findings are consistent with the hypothesis that IRBP, a 135K constituent of the interphotoreceptor matrix, is synthesized in vivo by a neuronal cell, specifically, the photoreceptors.
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牛神经视网膜合成和分泌的视黄醇结合糖蛋白的纯化和表征。
DOI：
--
发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Fong,SL;Liou,GI;Landers,RA;Alvarez,RA;Bridges,CD
通讯作者：
Bridges,CD
视网膜感光器间空间中存在可溶性感光器间视黄醇结合蛋白（IRBP）
影响因子：
5.6
作者：
B. A. Pfeffer;B. Wiggert;L. Lee;B. Zonnenberg;D. Newsome;G. Chader
通讯作者：
G. Chader
破伤风毒素：培养物中神经元的细胞表面标记
影响因子：
2.9
作者：
R. Mirsky;L. Wendon;Philippa Black;C. Stolkin;D. Bray
通讯作者：
D. Bray
影响因子：
4.4
作者：
A. Eisenfeld;Ann H. Bunr-Milam;P. V. Sarrhy
通讯作者：
A. Eisenfeld;Ann H. Bunr-Milam;P. V. Sarrhy
DOI：
10.1083/jcb.94.3.574
发表时间：
1982-09
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lander AD;Fujii DK;Gospodarowicz D;Reichardt LF
通讯作者：
Reichardt LF