A novel model of periventricular leukomalacia on mouse organotypic brain slice culture

Home/2016, Vol. 4, No. 2/A novel model of periventricular leukomalacia on mouse organotypic brain slice culture

Cell and Organ Transplantology. 2016; 4(2):188-193.
DOI: 10.22494/COT.V4I2.60

A novel model of periventricular leukomalacia on mouse organotypic brain slice culture

Tsupykov O. M.1,2, Lushnikova I. V.1, Nikandrova Y. A.1, Yatsenko K. V.1, Ustymenko A. M.2, Kyryk V. M.2, Butenko G. M.2, Skybo G. G.1,2
1Bogomolets Institute of Physiology NAS of Ukraine, Kyiv, Ukraine
2State Institute of Genetic and Regenerative Medicine NAMS of Ukraine, Kyiv, Ukraine

The creation of adequate in vitro and in vivo models of neural tissue injury is essential to assess the therapeutic effect of pharmacological agents and regenerative potential of various types of stem cells in diseases of the central nervous system. The aim of this work was to create a novel model of cerebral white matter lesions – periventricular leukomalacia (PVL) – on murine organotypic brain slice culture.
Materials and methods. The PVL model was developed on cultured organotypic mice brain slices subjected to oxygen-glucose deprivation (OGD) followed by addition of endotoxin lipopolysaccharide (LPS) in the culture medium. To analyze the degree of tissue injury within PVL simulation, we used spectrophotometric method for estimation of cytosolic enzyme lactate dehydrogenase (LDH) in the culture medium and immunohistochemical analysis of the slices using antibodies to Rip, GFAP and Iba-1 protein markers of oligodendrocyte, astroglia and microglia, respectively.
Results. It was shown that the combined effect of OGD and lipopolysaccharide resulted in a significant release of the cytosolic enzyme LDH in culture medium, decrease of Rip-immunoreactivity and a pronounced reactive astro- and microgliosis in murine organotypic brain slice culture.
Conclusions. Our model of PVL developed on cultured organotypic mice brain slices is novel and promising tool to study pathogenic mechanisms of cerebral white matter lesions and ways of neuroprotection in this pathology, including pharmacological agents and transplantation of stem cells.

Key words: periventricular leukomalacia; lipopolysaccharide; oxygen-glucose deprivation; organotypic brain slice culture

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Tsupykov OM, Lushnikova IV, Nikandrova YA, Yatsenko KV, Ustymenko AM, Kyryk VM, Butenko GM, Skybo GG. A novel model of periventricular leukomalacia on mouse organotypic brain slice culture. Cell and Organ Transplantology. 2016; 4(2):188-93. doi:10.22494/cot.v4i2.60


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