A novel method for co‑culture with Müller cells and microglia in rat retina in vitro

Li,Li; Qu,Chen; Wang,Fang

doi:10.3892/br.2014.370

A novel method for co‑culture with Müller cells and microglia in rat retina in vitro

Authors:
- Li Li
- Chen Qu
- Fang Wang
View Affiliations

Affiliations: Department of Ophthalmology, Shanghai Tenth People's Hospital Affiliated to Tongji University, Shanghai 200072, P.R. China
Published online on: October 13, 2014 https://doi.org/10.3892/br.2014.370
Pages: 25-27

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Microglia and Müller cells are glial cells of the retina and constitute a functional link between neurons and vessels. The aim of the present study was to introduce a novel method of co‑culture with Müller cells and microglia in rat retina. A camera was used to analyze all the cell changes. Immunofluorescence staining of glutamine synthetase and OX‑42 were used for the identification of Müller cells and microglial, respectively. On day 1, all the cell types from the retina were round or oval and floating in the medium. On the following days, microglial cells were adherent and proliferated. Müller cells stretched and quickly proliferated. On days 12‑15, microglial cells were floating in the medium. Following agitation, microglial cells became quickly detached from the flask walls, whereas Müller cells remained adherent. In conclusion, agitation is an effective way to separate microglial cells from Müller cells. The time of detachment and the speed of agitation are essential. Co‑culture with Müller cells and microglia in the retina is economical and useful for future methods in microglia and Müller cell research.

View Figures

View References

1	Reichenbach A, Wurm A, Pannicke T, Iandiev I, Wiedemann P and Bringmann A: Müller cells as players in retinal degeneration and edema. Graefes Arch Clin Exp Ophthalmol. 245:627–636. 2007. View Article : Google Scholar
2	Tezel G and Wax MB: The immune system and glaucoma. Curr Opin Ophthalmol. 15:80–84. 2004. View Article : Google Scholar : PubMed/NCBI
3	Forrester JV and McMenamin P: Immunopathogenic mechanisms in intraocular inflammation. Chem Immunol. 73:159–185. 1999. View Article : Google Scholar : PubMed/NCBI
4	Egensperger R, Maslim J, Bisti S, Holländer H and Stone J: Fate of DNA from retinal cells dying during development: uptake by microglia and macroglia (Müller cells). Brain Res Dev Brain Res. 97:1–8. 1996. View Article : Google Scholar : PubMed/NCBI
5	Roque RS, Imperial CJ and Caldwell RB: Microglial cells invade the outer retina as photoreceptors degenerate in Royal College of Surgeons rats. Invest Ophthalmol Vis Sci. 37:196–203. 1996.PubMed/NCBI
6	Dick AD, Carter D, Robertson M, et al: Control of myeloid activity during retinal inflammation. J Leukoc Biol. 74:161–166. 2003. View Article : Google Scholar : PubMed/NCBI
7	Bringmann A, Pannicke T, Grosche J, et al: Müller cells in the healthy and diseased retina. Prog Retin Eye Res. 25:397–424. 2006. View Article : Google Scholar
8	Walsh N, Valter K and Stone J: Cellular and subcellular patterns of expression of bFGF and CNTF in the normal and light stressed adult rat retina. Exp Eye Res. 72:495–501. 2001. View Article : Google Scholar : PubMed/NCBI
9	Harada T, Harada C, Kohsaka S, et al: Microglia-Müller glia cell interactions control neurotrophic factor production during light-induced retinal degeneration. J Neurosci. 22:9228–9236. 2002.
10	Saura J: Microglial cells in astroglial cultures: a cautionary note. J Neuroinflammation. 4:262007. View Article : Google Scholar : PubMed/NCBI
11	Chen H and Weber AJ: Expression of glial fibrillary acidic protein and glutamine synthetase by Müller cells after optic nerve damage and intravitreal application of brain-derived neurotrophic factor. Glia. 38:115–125. 2002. View Article : Google Scholar
12	Park LC, Zhang H and Gibson GE: Co-culture with astrocytes or microglia protects metabolically impaired neurons. Mech Ageing Dev. 123:21–27. 2001. View Article : Google Scholar : PubMed/NCBI
13	Zack DJ: Neurotrophic rescue of photoreceptors: are Müller cells the mediators of survival? Neuron. 26:285–286. 2000. View Article : Google Scholar
14	Tynan RJ, Naicker S, Hinwood M, et al: Chronic stress alters the density and morphology of microglia in a subset of stress-responsive brain regions. Brain Behav Immun. 24:1058–1068. 2010. View Article : Google Scholar : PubMed/NCBI
15	Tackenberg MA, Tucker BA, Swift JS, Jiang C, Redenti S, Greenberg KP, Flannery JG, Reichenbach A and Young MJ: Müller cell activation, proliferation and migration following laser injury. Mol Vis. 15:1886–1896. 2009.
16	Yu J, Huang X, Wu Q, Wang J, Yu X and Zhao P: Effect of A2A receptor antagonist (SCH 442416) on the mRNA expression of glutamate aspartate transporter and glutamine synthetase in rat retinal Müller cells under hypoxic conditions in vitro. Exp Ther Med. 3:803–806. 2012.PubMed/NCBI