J Korean Ophthalmol Soc.  2011 May;52(5):618-623. 10.3341/jkos.2011.52.5.618.

Effect of Creatine on the Survival of RGC-5 Cells under Serum Deprivation

• Kim JW
• Hong JH
• Kang SH
• Kim YY

• KMID: 2214583
• DOI: http://doi.org/10.3341/jkos.2011.52.5.618

Abstract

PURPOSE
To evaluate the protective effect of creatine on the survival of retinal ganglion cells after serum deprivation.
METHODS
RGC-5 cells were exposed to 5 mM creatine with serum-free media for 4 days. Cellular survival and mitochondrial respiratory activity were measured with MTT assay and resazurin assay, respectively. Degree of apoptosis was evaluated with vital staining using acridine orange/Hoechest 33342 and flow cytometric analysis using annexin/PI, respectively.
RESULTS
Creatine increased cellular survival of RGC-5 cells significantly after serum deprivation. Additionally, creatine increased mitochondrial respiratory activity and inhibited apoptosis of RGC-5 cells.
CONCLUSIONS
The energy precursor creatine increased survival of retinal ganglion cells after serum deprivation. Creatine could be relevant for the cytoprotection of retinal ganglion cells.

Keyword

Creatine; Cytoprotection; Retinal ganglion cells

MeSH Terms

Apoptosis
Creatine
Culture Media, Serum-Free
Cytoprotection
Oxazines
Retinal Ganglion Cells
Xanthenes
Creatine
Culture Media, Serum-Free
Oxazines
Xanthenes

Figure

• 

  Figure 1. Effect of creatine on the survival of RGC-5 cells after serum deprivation for 4 days. Addition of 5 mM creatine significantly increased cellular survival. * p < 0.05.

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  Figure 2. AO/HO double staining of RGC-5 cells with or without treatment of 5 mM creatine for 4 days. Normal control cell stains blue nuclei and brilliant red cytoplasm. In contrast, the apoptotic cell (arrows) shows yellow to orange-red nuclei, becoming indis-tinguishable from the cytoplasm. Addition of 5 mM creatine (A) revealed less apoptotic cells compared non-treated control (B) (×200).

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  Figure 3. Flow cytometric analysis of the effect of creatine on the survival of RGC-5 cells after serum deprivation for 4 days. Addition of 5 mM creatine significantly decreased cellular apoptosis. * p < 0.05.

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  Figure 4. Effect of creatine on the mitochondrial respiratory activity in the RGC-5 cells after serum deprivation for 4 days. Addition of 5 mM creatine significantly increased mitochondrial respiratory activity compared to non-treated controls.

Reference

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