Cell and Organ Transplantology. 2017; 5(1):28-32.
Protective effects of adipose-derived multipotent mesenchymal stromal cells of mice on periventricular leukomalacia model in vitro
Tsupykov O. M.1,2, Lushnikova I. V.1, Ustymenko A. M.2, Kyryk V. M.2, Nikandrova Y. A.1, Patseva M. A.1, Yatsenko K. V.1, Butenko G. M.2, Skibo G. G.1,2
1Bogomoletz Institute of Physiology NAS of Ukraine, Kyiv, Ukraine
2State Institute of Genetic and Regenerative Medicine NAMS of Ukraine, Kyiv, Ukraine
Periventricular leukomalacia (PVL) is a form of white matter lesions of the brain that results from hypoxic-ischemic injury and/or inflammation of nervous tissue, and is one of the causes of cerebral palsy. On PVL models in vivo, we have demonstrated neuroprotective effect of transplantation of adipose-derived multipotent mesenchymal stromal cells (MMSCs). However, the mechanisms, which realize neuroprotective effect of transplanted MMSCs, remain unexplored.
The aim was to assess the influence of adipose-derived MMSCs on cultured mouse brain slices at their contact co-culturing on PVL models in vitro.
Methods. Periventricular leukomalacia in vitro was modelled by a 30-minute oxygen-glucose deprivation (OGD) of mouse brain slices, followed by the addition of 100 ng/ml LPS in culture medium. For co-cultivation we used adipose-derived MMSCs obtained from mice FVB-Cg-Tg (GFPU) 5Nagy/J, transgenic for green fluorescent protein (GFP). The viability of cultured sections cells was evaluated by analysing the level of lactate dehydrogenase (LDH) in the culture medium. Probable MMSCs differentiation into neurons and glial cells was studied using immunohistochemical staining of slices using specific antibodies to neurons and oligodendrocytes (NeuN and Oligodendrocytes, respectively).
Results. Modelling of PVL in vitro on organotypic culture of brain slices led to a significant increase in level of cytosolic enzyme LDH in the culture medium. Co-cultivation of slices with MMSCs at PVL reduced the amount of this enzyme. Furthermore, it is shown that under conditions of PVL in vitro, MMSCs are able to differentiate into cells of nervous tissue.
Conclusions. Adipose-derived MMSCs have protective effect when they are co-cultivated with the mice brain slices on PVL model in vitro.
Key words: periventricular leukomalacia; lipopolysaccharide; brain organotypic tissue culture; multipotent mesenchymal stromal cellsFull Text PDF (eng) Full Text PDF (ua)
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Tsupykov OM, Lushnikova IV, Ustymenko AM, Kyryk VM, Nikandrova YA, Patseva MA, Yatsenko KV, Butenko GM, Skibo GG. Protective effects of adipose-derived multipotent mesenchymal stromal cells of mice on periventricular leukomalacia model in vitro. Cell and Organ Transplantology. 2017; 5(1):28-32. doi:10.22494/cot.v5i1.66