期刊文献+

成熟视网膜神经节细胞三维培养 被引量:4

Establishment of new culture system for retinal ganglion cells
下载PDF
导出
摘要 目的 :建立成熟视网膜神经节细胞的三维培养模型。方法 :将边长 5 0 0 μm的方形网膜片神经纤维层向下浸入 1 2孔培养板的胶原溶液层中 ,胶原溶液凝固后 ,培养孔内加入无血清 MEM培养液。相差显微镜观察生长情况。应用羊抗鼠 FITC- Thy- 1 .1单克隆抗体鉴定培养的神经节细胞突起。结果 :视网膜神经节细胞突起在凝胶中呈螺旋迂曲生长 ,可存活 2周以上。免疫组化荧光显色神经节细胞突起呈阳性反应。结论 :应用凝胶做支持物对成熟视网膜神经节细胞进行三维原代培养 ,是一种成功的。 ObjectiveTo establish a new culture system for retinal ganglion cells (RGCs). MethodsFour male Wistar rats aged 10 weeks, retina was isolated from the choroid, the pigment epithelium and adherent vitreous. The retinal explants from mid peripheral retinas were dissected into 16 pieces (approximately 0.5 mm×0.5 mm square). Four pieces of retinal tissues were embedded in the type I collagen gel with the nerve fiber layer downward on plastic dish. Each dish was pre coated with 10 mg·L -1 poly L lysine. The explants were cultured in serum free MEM and maintained at 37℃ in an incubator under a 5% CO 2/95% air atmosphere. The number of outgrowing neural protuberances was counted under phase contrast optics using inverted microscope. Immunohistochemical markers, monoclonal antibodies against neurofilament and Thy 1 1 were used to identify the nature of the outgrowing processes. ResultsUsing the three dimensional culture, the retinal explants were stably embedded just below the surface of the gel, and they could be cultured for 2 weeks or more. The outgrowing neural protuberances could elongate more freely into gel in the three dimensional culture than in the two dimensional one.ConclusionRGCs can be cultured sucecessfully in the three dimensional culture.
出处 《吉林大学学报(医学版)》 CAS CSCD 北大核心 2004年第2期236-238,共3页 Journal of Jilin University:Medicine Edition
基金 长春市科技发展计划项目 (0 1 - 1 1 2 5 36 )
关键词 视网膜神经节细胞 组织培养/方法 三维培养 retinal ganglion cells tissue culture/methods three dimensional culture
  • 引文网络
  • 相关文献

参考文献5

  • 1刘文文,徐萍,郭强苏,刘军,顾青,黄倩.培养的胚胎视网膜神经细胞Ca^(2+)分布与Ca^(2+)通道特征[J].中华眼底病杂志,2002,18(1):42-45. 被引量:1
  • 2Shatz CJ, Oleary DD. Repair and replacement to restore sight [J]. Arch Ophthalmol, 1993, 111: 472-477. 被引量:1
  • 3Raff MC, Fields KL, Hakomori SI, et al. Cell-type-specific markers for distinguishing and studying neurons and the major classes of glial cells in culture [J]. Brain Res, 1979, 174: 283-308. 被引量:1
  • 4Leifer D, Lipton SA, Barnstable CJ, et al. Monoclonal antibody to Thy-1 enhances regeneration of processes by rat retinal ganglion cells in culture [J]. Science, 1984, 224: 303-306. 被引量:1
  • 5Montague PR, Friedlander MJ. Expression of an intrinsic growth strategy by mammalian retinal neurons [J]. Proc Natl Acad Sci USA, 1989, 86: 7223-7227. 被引量:1

二级参考文献5

  • 1Satoh H,Aoki K,Watanabe SI,et al.L-type calcium channels in the axon terminal of mouse bipolar cells[].NeuroReport.1999 被引量:1
  • 2Philippov PP.Phototransduction and calcium[].Membrane and Cell Biology.2000 被引量:1
  • 3Wu D,Zhu PH.Caffeine-sensitive Ca2+ stores in carp retinal bipolar cells[].NeuroReport.1999 被引量:1
  • 4Payne R,Demas J.Timing of Ca2+ release from intracellular stores and the electrical response of Limulus ventral photoreceptors to dim flashes[].The Journal of General Physiology.2000 被引量:1
  • 5Nawy S.Regulation of the bipolar cell mGluR6 pathway by Ca2+[].The Journal of Neuroscience.2000 被引量:1

同被引文献109

引证文献4

二级引证文献5

;
使用帮助 返回顶部