Evaluation of the effectiveness of stem cell therapy with use of adult multipotent neural crest-derived stem cells for modeled adrenal-induced glaucoma treatment

Abstract

The aim of a study was to assess the morphological changes of retina structure after modeled adrenal-induced glaucoma and treatment effect of the postnatal cultured multipotent neural crest-derived stem cells (NC-MSCs) delivered by different routes. Glaucoma condition was made in Wistar rats (males, aged 10-12 months) by intraperitoneal injection of 0.18% solution of adrenaline tartrate in a dose ranged 10 to 15 µg per 100 g of body weight. There were 20 injections through 6 weeks. Measurement of intraocular pressure (IOP) was carried with use of applanation tonometer. Adult NC-MSCs were isolated from the bulge region of whisker follicle by explant methods. Adult NC-MSCs were expanded in vitro and then characterized by immunocytochemistry, flow cytometry and directed differentiation assay into NC-derived cell types. Cell transplantation modes: native NC-MSCs, i/v (5×106 ); retro- and parabulbar injections of 0.5×106 native and committed NC-MSCs (CNTF treatment, 5 days). Retina and optic nerve histomorphometric analysis was done on H&E stained sections. NC-MSCs have a phenotype of nestin+ p75+ Sox10+ cytokeratin- . Following lineage commitment cells become GFAP+ with morphology of Muller glia precursor cells. IOP in rats before modeling was 7-8 mmHg. During the injection of AT the IOP was increased in all experimental animals from 15 to 20 mmHg. Adrenal-induced glaucoma 1 month after modeling led to the development of stable through 3 months glaucoma characteristic dystrophic changes. NCMSCs transplantation induced positive changes for all cell delivery methods. Positive effect of the NC-MSCs administration was observed in terms of swelling regression, retina and optic nerve restoration. Retrobulbar mode had the most prominent effect on the recovery of ganglionic layer, restoration of the structure of inner and outer nuclear layers and layer of rods and cones. Further studies of mechanisms (trophic vs structural integration) involved in retina recovery should be done.