Isolation, characterization and establishment of an equine retinal glial cell line: a prerequisite to investigate the physiological function of Müller cells in the retina.
Abstract: Retinal Müller glial cells are of vital importance for maintaining a physiological environment within the retina. To this end, they provide highly specialized physiological properties to support neurons in structure, nutrition and metabolism. The purpose of this study was to isolate Müller cells from the equine retina, determine their characteristics and subsequently establish a stable equine Müller cell line (eqMC) that will provide a prerequisite for investigations on their physiological properties. Dissociated retinal cells were obtained from equine retinas by a papain digestion technique followed by trituration and a cell attachment method by which pure Müller cell cultures were achieved. Morphological examination was performed using phase-contrast microscopy, and further characterization of different subcultures was accomplished by immunocytochemistry. Cells of passage 1 showed distinct signals for glutamine synthetase and vimentin, whereas glial fibrillary acidic protein expression was almost absent. Characteristic expression patterns remained unaltered in all subcultures. Furthermore, cultured Müller cells stably expressed the microfilament alpha-smooth muscle actin, the proliferation marker Ki67 and the membrane channels Kir4.1 and aquaporin 4. The present study introduces the eqMC-7 that will facilitate studies investigating the physiological role of Müller cells within the equine retina.
© 2011 Blackwell Verlag GmbH.
Publication Date: 2011-04-30 PubMed ID: 21535230DOI: 10.1111/j.1439-0396.2011.01147.xGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article is about the isolation and characterization of Müller cells from the equine retina in order to establish a stable cell line for studying their physiological functions.
Isolation and Culturing of Müller Cells
- The researchers isolated Müller cells from the retina of horses (equines).
- A chemical called papain was used to digest the retinal tissue and dissociate the cells, which were then gently crushed (triturated) and allowed to attach to a surface to grow and multiply.
- Through these processes, the researchers were able to establish a culture of pure Müller cells.
Characterization of Müller Cells
- The isolated cells were examined under a microscope for their physical characteristics.
- The researchers also used a technique called immunocytochemistry to identify specific proteins in the cells, which can provide insight into the cells’ functions.
- They found that the cells produced two types of proteins (glutamine synthetase and vimentin) often seen in Müller cells, but not another common protein (glial fibrillary acidic protein).
- The expression patterns of these proteins stayed the same even as the cells were grown and multiplied in subsequent cultures (subcultures).
Establishment of the eqMC-7 Cell Line
- The researchers established a stable cell line (called eqMC-7) from the isolated Müller cells.
- This cell line consistently produced certain molecules and proteins associated with Müller cells, including a type of protein fiber (alpha-smooth muscle actin), a marker of cell division (Ki67), and two types of channels in the cell membrane (Kir4.1 and aquaporin 4).
Implications of the Study
- The establishment of the eqMC-7 cell line will enable further research on the physiological functions of Müller cells in the equine retina.
- This could help improve understanding of retinal health and disease, potentially leading to new treatments for retinal disorders in horses and other animals.
Cite This Article
APA
Eberhardt C, Amann B, Stangassinger M, Hauck SM, Deeg CA.
(2011).
Isolation, characterization and establishment of an equine retinal glial cell line: a prerequisite to investigate the physiological function of Müller cells in the retina.
J Anim Physiol Anim Nutr (Berl), 96(2), 260-269.
https://doi.org/10.1111/j.1439-0396.2011.01147.x Publication
Researcher Affiliations
- Department of Veterinary Sciences, Institute of Animal Physiology, Ludwig-Maximilians University, München, Germany.
MeSH Terms
- Actins / genetics
- Actins / metabolism
- Animals
- Biomarkers
- Cell Line
- Gene Expression Regulation / physiology
- Horses / physiology
- Membrane Proteins / genetics
- Membrane Proteins / metabolism
- Neuroglia / cytology
- Neuroglia / physiology
- Retina / cytology
Citations
This article has been cited 7 times.- Kang S, Wohl SG. Primary Cell Cultures to Study the Regeneration Potential of Murine Müller Glia after MicroRNA Treatment.. J Vis Exp 2022 Mar 28;(181).
- Schmalen A, Lorenz L, Grosche A, Pauly D, Deeg CA, Hauck SM. Proteomic Phenotyping of Stimulated Müller Cells Uncovers Profound Pro-Inflammatory Signaling and Antigen-Presenting Capacity.. Front Pharmacol 2021;12:771571.
- Lorenz L, Hirmer S, Schmalen A, Hauck SM, Deeg CA. Cell Surface Profiling of Retinal Müller Glial Cells Reveals Association to Immune Pathways after LPS Stimulation.. Cells 2021 Mar 23;10(3).
- Degroote RL, Deeg CA. Immunological Insights in Equine Recurrent Uveitis.. Front Immunol 2020;11:609855.
- Deeg CA, Amann B, Lutz K, Hirmer S, Lutterberg K, Kremmer E, Hauck SM. Aquaporin 11, a regulator of water efflux at retinal Müller glial cell surface decreases concomitant with immune-mediated gliosis.. J Neuroinflammation 2016 Apr 23;13(1):89.
- Lin X, Fang D, Zhou H, Su SB. The expression of Toll-like receptors in murine Müller cells, the glial cells in retina.. Neurol Sci 2013 Aug;34(8):1339-46.
- Deeg CA, Eberhardt C, Hofmaier F, Amann B, Hauck SM. Osteopontin and fibronectin levels are decreased in vitreous of autoimmune uveitis and retinal expression of both proteins indicates ECM re-modeling.. PLoS One 2011;6(12):e27674.
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