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  • Isolation and characterization of human optic nerve head astrocytes and lamina cribrosa cells.

Isolation and characterization of human optic nerve head astrocytes and lamina cribrosa cells.

Experimental eye research (2020-06-12)
Navita N Lopez, Abbot F Clark, Tara Tovar-Vidales
要旨

The lamina cribrosa is the initial site of glaucomatous injury. Pathological changes to the lamina cribrosa include posterior displacement of the lamina cribrosa, loss of trophic support, and remodeling of the extracellular matrix. Optic nerve head (ONH) astrocytes and lamina cribrosa cells synthesize extracellular matrix proteins to support and maintain the lamina cribrosa under physiological conditions. During glaucoma, these cells respond to mechanical strain and other stimuli, which leads to pathological remodeling of the ONH. Although ONH astrocytes and lamina cribrosa cells have been previously cultured, there is no well-accepted, straightforward technique to isolate both cell types from a single dissected human ONH. To better understand the pathophysiology of glaucoma, we obtained and cultured lamina cribrosa explants from human donor eyes. Initially, cells that grew out from the explant were ONH astrocytes and lamina cribrosa cells. Using a specialized medium, we isolated pure populations of lamina cribrosa cells and ONH astrocytes. ONH astrocytes expressed glial fibrillary acidic protein (GFAP). Lamina cribrosa cells expressed alpha-smooth muscle actin (α-SMA), but were negative for GFAP. This method of ONH cell isolation and cell-culture will provide a technique to better understand the molecular and cell-specific changes in glaucomatous damage to the ONH.

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製品内容

Sigma-Aldrich
ダルベッコリン酸緩衝生理食塩水, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
ペニシリン-ストレプトマイシン, Solution stabilized, with 10,000 units penicillin and 10 mg streptomycin/mL, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
抗ラミニン ウサギ宿主抗体, 0.5 mg/mL, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
抗アクチンα-平滑筋-FITC抗体、マウスモノクローナル マウス宿主抗体, clone 1A4, purified from hybridoma cell culture
Sigma-Aldrich
栄養混合物 F-10ハム, With sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
モノクロナール抗神経細胞接着分子 マウス宿主抗体, clone NCAM-0B11, ascites fluid