The technology of obtaining multipotent spheroids from limbal mesenchymal stromal cells for reparation of injured eye tissues.

It is known that stem and progenitor cells open new possibilities for restoring injured eye tissues. Limbal eye zone, formed mainly by derivatives of neural crest, is the main source of stem cells for regeneration. The current study considers development of innovative technology for obtaining 3D spheroids from L-MMSC. It was shown that under 3D conditions L-MMSC due to compactization and mesenchymal-epithelial transition self-organize into cellular reparative modules. Formed L-MMSC spheroids retain and promote undifferentiated population of stem and progenitor limbal cells, as supported by expression of pluripotency markers - Oct4, Sox2, Nanog. Extracellular matrix synthetized by cells in spheroids allows retaining the functional potential of L-MMSC that are involved in regeneration of both anterior and, probably, posterior eye segment.

[Experience of culturing anterior epithelial corneal cells from human eye ball].

Adult corneal epithelium is often exposed to environmental stress, injured and repaired by limbal stem cells. Injury of corneal epithelial layer leads to reduction of visual clarity and loss of vision. Recently it was shown that epithelial layer also contains stem cells. Obtaining cell culture of corneal epithelium will allow understanding mechanisms of cell behavior and differentiation, their metabolism and reaction on environmental stress in health and disease. Moreover, cultured corneal epithelial cells can be considered as a promising material for constructing bioartificial cornea. The aim of this study was to isolate cells of anterior corneal epithelium from human donor cornea and to study their morphological and functional characteristics in vitro. The results of our study showed the possibility of culturing epithelial cells in vitro. The observed changes in cell morphology, their flow growth character as well as active proliferation and up-regulation of mesenchymal markers expression, indicate, in our opinion, epithelial-mesenchymal transition taking place in long-lasting culture of human anterior corneal epithelial cells. The obtained cultures can be used for further studies of pathological processes taking place in cells during drugs testing or controlling the phototoxic effect of different types of emission.

Evaluation of the safety of laser-induced fluorescence on the model of human diploid cell culture.

The possibility of using L-68 diploid cell culture as a model for evaluation of the safety of laser-induced fluorescence stimulated by Kr-F eximer laser (lambda=248 nm) is proven. Laser irradiation of human diploid fibroblasts in a dose of 300 mJ/cm2 and higher led to changes in cell morphology, decreased their proliferative activity and viability, and induced accumulation of lipid peroxidation products (carbonyl compounds) in the nutrient medium. Irradiation doses below 100 pulses did not modify cultural and morphological characteristics of cells.

Cell-based therapy of chronic degenerative diseases of the central nervous system.

The traditional methods of pharmacotherapy of the degenerative diseases of the central nervous system do not frequently allow one to achieve the desired clinical effect. The fundamentally new approach for the treatment of severe neurological diseases is provided by the methods of biological medicine, in particular, transplantation of a complex of fetal tissues. Cell-based therapy was used to treat patients with multiple sclerosis; ante-, intra- and postnatal lesions; consequences of hemorrhagic and ischemic apoplexies; neuritis of facial nerve; sclerosis; Parkinson's disease; Alzheimer's disease; epilepsy and other types of pathologic process. The source material for obtaining a suspension of cells was the fetuses of allogenic origin. The suspension of brain cells in amounts of up to 1.5 x 10(8) cells and vitality not less than 40% was administered to the patients into liquor spaces using the method of endolumbar puncture. The total number of transplantations was 1900. Practically in all the cases FT was tolerated well. Positive clinical and immunologic changes were observed in the majority of the patients, thus, remission induction (in the patients with the progressive course of multiple sclerosis) for a period over 12 months was registered in 87.5% of the cases. Noteworthy that considerable changes were observed in immunograms: depression of antibody levels to brain-specific proteins, native and denatured DNA; quantitative and qualitative improvement of lymphocyte subpopulation indices, positive changes in the immunoregulatory index. Clinically, in 69% of the cases there was an improvement in more than one neurological defect and a change in the values of the Kurtzke scale towards a decrease by 2-3 points. The conduct of cell therapy with the MS patients under the acute process conditions after liquorosorption allowed the arresting of clinical manifestations and the creation of preconditions for further restoration. The retrobulbar transplantations provided a quick arrest of the retrobulbar neuritis clinical symptoms and in one case an almost complete restoration of vision in the patient with amaurosis (blindness). The remission duration has a marked direct dependence on the number of courses of endolumbar transplantations. Thus, the method of cell therapy with the use of human tissue transplantations is safe and can be used for different neurodegenerative lesions of the central nervous system. The high efficacy of the method suggests the possibility and necessity of using this method as an alternative of classical pharmacological therapy. An important element of cell therapy is the control after the state of the patient's immunity system.