Mesenchymal stem cells from human umbilical cord (1x10(6)) are able to correct neurodegenerative changes in the spinal cord of rats with chronic experimental allergic encephalomyelitis under the suboccipital administration. Cells cryopreserved in solution with 3 % dimethyl sulfoxide, 15 % ethylene glycol, 10 % sucrose, 12 % trehalose, 60 % fetal calf serum, slow the buildup of pathological changes at day 35; cryopreserved in solution with 4 % dimethyl sulfoxide, 6 % trehalose, 90 % fetal calf serum, - at days 35-60; cryopreserved in solution with 10 % dimethyl sulfoxide, 90 % fetal calf serum, - have no effect. Selection of optimal cryoprotectants to save the biological properties of these cells and working out the modes and ways of their administration remains relevant.

In the chronic course of experimental allergic encephalomyelitis neurodegenerative changes in the brain tissue of rats exacerbate within 2 months. Suboccipital administration of mesenchymal stem cells obtained from human umbilical cord (1x106) and cryopreserved in the solution containing 3% dimethyl sulfoxide, 15% ethylene glycol, 10% sucrose, 12% trehalose, and 60% fetal calf serum inhibits these changes in hippocampus (day 35); administration of those cryopreserved in the solution containing 4% dimethyl sulfoxide, 6% trehalose, and 90% fetal calf serum inhibits these changes in both hippocampus (days 35- 60) and brain cortex (day 60); administration of these cells cryopreserved in the solution containing 10% dimethyl sulfoxide and 90% fetal calf serum slows down the changes in hippocampus (days 35-60) and partly recover the neuronal component in the brain cortex (day 60).

This review presents recent data on the biological properties of cancer stem cells in brain gliomas. The possibilities of using of experimental methods of cell culture and immunohistochemical identification of tumor stem cells as a part of neurospheres (tumorospheres) are shown. Methodological features of neurospheres culture are described.Literature data about the dependence of neurospheres formation activity in the culture on the degree of anaplasia of original glioma tissues are presented considering the disease outcomes in neuro-oncological patients. Possibilities of the practical use of cultured tumorospheres in the study of selective impact of antiblastic drugs on tumor stem cells in gliomas are described.

The increased interest in the application of lasers in neuro-oncology prompted us to present our experience of using the laser technologies in the treatment of cerebral gliomas. The aim of the study was to evaluate the clinical efficacy of image-guided laser surface thermal therapy (LSTT) and its influence on survival of patients with glioblastoma (GBM).

The review summarizes the current concepts of cell-tissue and molecular features of development of demyelinating processes in the central nervous system related to multiple sclerosis and its animal model – allergic encephalomyelitis. An analysis of recently published studies of this pathology, carried out with light and electron microscopy and immunohistochemical and molecular genetic methods, is given. New methodological approaches to the study of the pathomorhological aspects of demyelinating disorders allowed receiving in-depth understanding of the etiology and mechanisms of demyelination processes in the brain and spinal cord tissues at the cellular level and identifying the ways to develop effective modern methods of pathogenetic treatment of these diseases using cell therapy.

The aim of the work was to study the impact of fetal rat brain cell supernatant (FRBCS) on the expression of transforming growth factor β1 (TGF-β1) and p53 in C6 cells of rat glioma in vitro.MATERIALS AND METHODS:FRBCS was obtained from suspensions of fetal rat brain cells on the 14th (E14) day of gestation. C6 glioma cells were cultured for 48 h in the presence of FRBCS or FRBCS + anti-TGF-β1 monoclonal antibody. Immunocytochemical staining for TGF-β1 and p53 was performed.RESULTS:The proportion of TGF-β1-immunopositive tumor cells in C6 glioma cultures was statistically significantly higher than in the control cell cultures of normal fetal rat brain. FRBCS reduced the proportion of TGF-β1-immunopositive tumor cells and increased the proportion of p53-immunopositive cells in C6 glioma cultures. In cells cultured with FRBCS + anti-TGF-β1 monoclonal antibody, the above effects of FRBCS were abrogated.CONCLUSION:The obtained results suggest that TGF-β1 seems to be responsible for decrease in TGF-β1 expression and increase in p53 expression in C6 glioma cells treated with FRBCS.

Objective – to improve treatment results in patients with periventricular oligoastrocytomas. Materials and methods. Oligoastrocytoma (OA) WHO Grade II was diagnosed in 16 (19.8 %) patients, OA WHO Grade III – in 65 (80.2 %). Diagnostic included the analysis of CT, MRI, SPECT in native modes and with contrast enhancement. As a source of laser radiation, the semiconductor laser «Lika-Chirurg» (wave length 0.808 and 1.47 p,m, 30 and 7 W) was used. The navigational support for surgical interventions was carried out using the Medtronic StealthStation TREONPlus (Medtronic, USA) surgical navigation system. Results. In 27 (33.3 %) patients with OA was a predominance of oligodendroglial component, in 28 (34.6 %) – astrocytic component, in 26 (32.1 %) – an equal representation of both components cells. The surgical treatment was the following: total removal – in 55 (67.9 %) patients, subtotal removal – in 26 (22.1 %). In the presurgery period quality of life with KRI > 80 % was assessed in 34 (41.9 %) patients, in postsurgery period – in 77 (95.1 %). Conclusions. Differential use of laser technologies and navigation support, taking into account the OA histology and topography, allows to optimize surgical tactics and totally remove a tumor without neural structuresinjury.

In order to standardize the model of experimental allergic encephalomyelitis (EAE) with development of chronic remitting course of disease a study of the different doses Freund’s Complete adjuvant (AF) effects and encephalolithic mixture on the degree of severity of behavioral reactions in the test «open field» of rats and the process of spinal cord demyelination was conducted. When testing rats in the «open field» with inducted EAE by Freund’s adjuvant single dose, on the 12th day, the inhibition of tentative-research and activation of emotional activity were revealed. Comparative pathomorphological analysis of the spinal cord of rats histostructure features with EAE on 35th day showed that the use of both double and triple doses of Freund’s adjuvant for induction provides the development of a more widespread demyelinating process in comparison with the single dose of Freund’s adjuvant. The use of a single dose of Freund’s adjuvant is optimal for obtaining an effective model of experimental allergic encephalomyelitis in rats with chronic recurrence.

One of the directions of cell therapy being developed for brain gliomas is the use of the neurogenic stem and progenitor cells (NSCs/NPCs). There are data on the anti-tumor and immunomodulating properties of the NSCs/NPCs the mechanisms of which were not disclosed yet. One of the potential targets for tumor therapy is the transforming growth factor β (TGF-β1) which is thought to be one of the key molecules in the regulation of proliferation, differentiation and cell survival or apoptosis. In the view of available information about the possibility of TGF-β1 production by the mammalian multipotent NSCs/NPCs, the aim of this work was to study the TGF-β1-positive cells in the dynamics of cultivation of fetal brain neurogenic cells as a potential source of anti-tumor or immunomodulating effects of these cells.Material and methods. The fetal rat brain cells on 14th (E14) day of gestation were used as the source for cultivation in standard conditions (DМЕМ + 1 % fetal bovine serum) and studied on the 2nd and 37thday by morphometry and immunocytochemistry.Results. In the fetal rat brain cell cultures, the TGF-β1-positive cells made 22.04 ± 2.33 % and the nestin-positive cells made 49.16 ± 10.60 % of the total cells number. The morphometric parameters of TGF-β1-positive cells exceeded the corresponding values of negative cells (average values of cross-sectional areas of the cytoplasm, cross-sectional areas of the nucleus, nuclear-cytoplasmic ratio). During cultivation the relative amount of TGF-β1-positive cells was slightly decreased 15.27 ± 9.80 % (p = 0.7) and their sizes were increased. On the 37th day of cultivation the sizes of TGF-β1-positive and their nuclei were smaller in the comparison with the TGF-β1-negative cells.Conclusions. The presence of TGF-β1 expression by part of neurogenic cells of fetal rat brain (E14) in vitro was found, which persisted throughout cultivation (~5 weeks). Significant quantitative differences of morphometric parameters of TGF-β1-positive and negative cells were detected.

Relevance of the topic. A promising tool for restoring the spinal cord function is the application of tissue neuroengineering. The approbation of its techniques is possible on qualitative spinal cord injury models that makes the transplantation of solid neuroengineered matrices feasible.Objective. To optimize and test a model of lower thoracic rat’s spinal cord hemisection.Materials and methods. 2 groups of experimental animals: 1 — mature animals (3–6 months, n=40) and 2 — young animals (1 month, n=32), the injury — left-side spinal cord hemisection (LHS) at Т11 level; monitoring of hindlimbs’ function (BBB scale), pathomorphological study.Results. LHS provides simultaneous display of a slight, moderate and severe spinal cord injury (the correlates, respectively, are: the function deficit of a contralateral hindlimb [CH] and an ipsilateral hindlimb [IH] among the animals with better and worse recovery indicators). As of the 11th week of the observation a function indicator was 3,2±0,58 points under BBB scale and 5,31±0,79 (p<0.05) in group 2, which indicates a significant fullness of the intersection of all descending fibers. Mortality in group 1 at the stage of intervention and in the acute phase of trauma consists 25%, in the remote phase — 15%, including 5 percent of animals with bilateral injury. At full bilateral intersection mortality within 10 days with relevant conditions of detention comprises 100 percent.Conclusion. LHS model is technically simple, easy to be reproduced, has low mortality under the condition of deep ipsilateral spinal cord function deficit, provides for three simultaneous options of spinal cord injury, is adapted for testing the neuroengineering tools.

Background. Penetrating spinal cord injury with a foreign body in the spinal canal is one of the most common spinal cord injuries during wartime; the experimental reproduction of particular elements of complex interaction between a foreign body and the spinal cord is complicated.Objective. To examine clinical and pathomorphological features of the model of this type of spinal cord injury.Materials and methods. Animals: albino male rats (5.5 months, 300 grams, inbred line, the original strain is Wistar); experimental groups: basic (spinal cord injury + immediate homotopical implantation of a fragment of the microporous hydrogel – a foreign body [n=10]); comparison groups (spinal cord injury [n=16], spinal cord injury + immediate homotopical implantation of chemically identical macroporous hydrogel NeuroGel™ [n=20]). Model of injury: left-side spinal cord hemisection at ТXI level; monitoring the function of hind legs — the BBB scale; pathomorphological study: conventional histological techniques, transmission electronic microscopy.Results. Compression of the spinal cord by biologically compatible foreign body significantly worsens the course of the regeneration process; during the first 8 weeks the hind ipsilateral leg function indicator (HI LFI) in animals was the lowest one — (1.30±0.94) points by BBB scale; during the 3rd–4th month HI LFI increases to 2.35±0.95 points by BBB scale, which is likely due to the change in the form of a foreign body and its utilization, decrease of the pressure on the spinal cord. On the 24th week of the follow-up HI LFI was (8.45±0.92) points (in NeuroGelTM group) compared with (2.35±0.95) points by BBB scale (in the group with a foreign body). During the experiment a foreign body, unlike the fragments of the NeuroGelTM, was not integrated into the tissue of the spinal cord, was surrounded by a thick fibrous capsule, hardly infiltrated by tissue component. Morphological picture in the contra-lateral part of the spinal cord at the level of injury did not change.Conclusion. The model satisfactorily a mechanical component of a foreign body effect on the spinal cord tissue, presents the picture of post-traumatic syndrome of spasticity; reducing the spinal cord compression even at the late period of injury significantly improves the regeneration process.

We estimated the effects of a rat fetal neurogenic cell supernatant (RFNCS) on proliferative activity of cultured cells of rat brain glioma (C6 cell line). The RFNCS was obtained from a cell suspension of the brains from rat fetuses (gestation day 14, E14). We added 0.10 mg/ml of RFNCS to the culture medium of experimental cultures and incubated the latter for 48 h. Immunocytochemical staining with respect to the Ki-67 proliferation marker was performed using rabbit monoclonal antibodies against this protein. In glioma C6 cell cultures, we observed degenerating and necrobiotically modified tumor cells with rounded cell bodies and reduction of the processes. Under the action of RFNCS, the mean density of the cells in 10 test fields of vision in experimental cultures (0.04 mm 2 ) became significantly smaller than that in the control (332.0 ± 36.0 vs. 569.5 ± 70.5; P = 0.00026); the mean value of the nuclear/cytoplasmic ratio in malignant cells mildly (insignificantly) decreased (0.28 ± 0.01 vs. 0.32 ± 0.02; P = 0.64) and the share of cells with multiple nucleoli was more than two times smaller (3.53 ± 0.33% and 7.97 ± 0.25%, respectively; P = 0.053). The mitotic index of cultured malignant cells subjected to the action of RFNCS became four times lower (1.10 ± 0.04% vs. 4.90 ± 0.09%; P = 0.009), while the share of malignant cells immunopositive with respect to Ki-67 decreased (10.47 ± 0.91% vs. 27.86 ± ± 2.91%; P = 0.0015). The observed antiproliferative effect of RFNCS confirms the possibility and expedience of the development of complex pathogenetic therapy for malignant tumors of the brain using preparations obtained from fetal neurogenic cells.

Simulation model of open penetrating wound of a spinal cord (SC) with its durable com- pression by biocompatible foreign body, was tasted. Experimental animals - mature male rats (inbred line, descendent from a Wistar breed); the trauma simulation model - a left- sided transsection of the SC half on the Txi level; experimental groups: the main (a SC trau- ma + homotopic implantation of microporous hydrogel fragment - foreign body [n=10]), groups of comparison (the SC trauma [n=16]; the SC trauma + homotopic implantation of chemically identical analogue of hydrogel - NeuroGelTM [n=20]). A SC compression by a foreign body worsens the regeneration process course essentially: during first 2 mo the function index of a hind ipsilateral extremity in experimental animals of the main group was the lowest in the experiment - (1.30 ? 0.94) points in accordance to BBB scale, during 3 - 4 mo - the function index had enhanced trustworthy - up to (2.35 ? 0.95) points in accor- dance to BBB scale, what is connected with lowering of a local pressure on a SC tissue due to change of the foreign body form and volume. In 24 weeks the function index of hind ipsi- lateral extremity had constituted (8.45 ? 0.92) points - while application of NeuroGeTM and (2.35 ? 0.95) points - of the foreign body; the tissue processes in the implants localization zone had differed essentially. The tasted simulation model reproduces satisfactory a mechanical component of the foreign body impact on a SC tissue. The SC compression reduction, even in a late follow-up period, had improved the conditions and results of recenerative process essentially.

Keywords: progenitor neurogenic cells, rat fetal brain, supernatant, glioma 101.8, cytotoxic index, mitotic index

Introduction. Concerning the relevance of estimation of radiation therapy (RT) efficiency in adults with cerebellar medulloblastoma, histological features of therapeutic pathomorphosis in tumors tissue after RT studied.Materials and methods. Comparative aspects of medulloblastomas histological structure investigated in 24 adult patients before and after RT using biopsy material at first and repeated operations.Results. Based on assessment of cell-tissue changes severity in medulloblastoma in adults after RT we installed individual differences of sensitivity to RT in most of them.Conclusions. Standard RT course in adults with medulloblastoma after surgery induces in tissue of most “treated” tumors cytotoxic and cytodestructive effect that correlates with disease-free period prolongation.

Cellular and molecular similarities between brain tumor stem cells (BTSCs) and normal neurogenic stem cells (NSCs) motivate the search for new methods of treatment of malignant glioma using NSCs. CD133 molecule could be one of the most typical markers of BTSCs and considered as a target for therapy of brain tumors.The aim of this study was to evaluate the effect of rat neurogenic cells supernatant (NCsS) on the content of CD133+ cells in glioma C6 cell cultures.Materials and methods. The cells of rat brain glioma C6 were used as the source for the cultivation; for comparative assessment of tested compound impact on the intact nervous system the fetal rat brain cells on 14th (E14) day of gestation were used. The study was performed in control cultures under standard culture conditions without NCsS adding and tested cultures with adding NCsS (0.10 mg/ml of protein) for 48 hours. NCsS was received from suspensions of rat brain neurogenic cells (E14).Results. CD133-positive cells were 12.05 ± 4.77 % of the total number of cells in C6 glioma culture and 37.36 ± 12.33 % of the total number of cells in fetal rat brain culture. CD133-positive cells had a smaller size than negative cells (average values of cross-sectional area of cells and nucleus) and greater nuclear-cytoplasmic ratio. The cell and nucleus sizes of CD133-positive cells in cell cultures of fetal rat brain were twice larger than sizes of such cells in cultures of glioma C6.Under the conditions of NCsS for 48 hours the reducing in the number of CD133-positive cells in rat glioma C6 cell cultures (2.88 ± 0.41 %) and lack of such effects in cell cultures of fetal rat brain (E14) were found.Conclusion. The morphological differences of CD133-positive cells in glioma C6 cultures and in cell cultures of fetal rat brain (E14) were detected. The decrease of CD133-positive cells in glioma C6 cells culture under the influence of neurogenic cells supernatant was shown.

Introduction. Despite intensive research of malignant gliomas in the world, today significant progress in their treatment are not attained by invasiveness and high retsydyvnist gliomas. The combination in treatment of cancer surgery, radiation and chemotherapy is the gold standard, but does not ensure its effectiveness. One alternative approaches to solve this problem is to use neurogenic stem cells and progenitor cells. It is known that neural progenitor cells exhibit antitumor properties in mice and rats. Multipotent neural progenitor cells of human, rat and mouse expressing both proinflammatory and suppressor cytokines. However, the mechanism of antitumor properties of neurogenic progenitor cells remains unclear. According to previous studies, fetal nerve cells of rats 18-20 days gestation exhibit antitumor effect in experimental glioma cells in vitro and 101.8 in vivo at heterotransplantatsiyi joint capsule in the kidneys of mice. Shown anti-tumor effect of the supernatant of fetal rat brain cells in rats with glioma 101.8 in vivo. The purpose of the study was to evaluate the effect of rat fetal neurogenic cells supernatant (RFNS) on cultured cells of rat glioma C6. Materials and methods. RFNS from suspensions of neurogenic cells of rat brain of 9-th (E9) and 14-th (T14) days of gestation (0,01, 0,10 mg/ml) were added tо cell suspensions of rat glioma C6; after 24 h of incubation cell suspensions were analyzed for number of viable cells, cytotoxic index (CІ) was calculated. RFNS (0,01, 0,10 mg/ml) were added tо primary cultures of rat glioma C6, after 24 and 48 h of incubation cytological preparations were analized and mitotic index was determined. The supernatant neural cells (NCC) received suspensions of neurogenic rat brain cells at 9 (E9) and 14 (E14) day of gestation. Results. Supernatants fetal rat brain both studied gestation (E9, E14) revealed a cytotoxic effect on all samples C6 glioma cells in short-term cultures. The level of exposure cytotoxic intensified with increasing duration of incubation of cells with supernatants. With increasing concentration of NCC (E9) from 0.01 to 0.10 mg/ml in the suspensions THESE C6 glioma cells increased and reached (37,61±2,57)% and (50,12±4,55)%, respectively, at 24 hours. and 48 hours. incubation. At 4-5 day culture experienced a gradual rozryhlennya mikroeksplantativ cell to form a monolayer of tumor cells rozroschen unipolar, triangular or rhomboid shape with fairly long spikes. In these areas crop growth zone were more common areas sitkopodibnoyi architectonics due to the formation of intercellular communications. In subsequent observation period (8-9 days) in cultures of C6 glioma There was a further dilution of cell arrays zone growth due to spontaneously share desquamation of dead cells appeared significant number of apoptotic cells, reflecting the spontaneous death of cultured cells. Necrobiotic processes spontaneous death covered the vast number of cells zone growth remained preserved some areas sitkopodibnyh structures and cellular sferoyidni microaggregates, surrounded by a monolayer of epithelial cell growths undifferentiated type. Conclusions. 1. The supernatant neural cells fetal rat brain different terms of gestation and antymitotychnyy found cytotoxic effects on cultured C6 glioma cells, which intensified with increasing gestational age rat brain, lengthening the duration of incubation was dose-dependent. 2. Primary cultures of rat brain gliomas is adequate experimental model for evaluation of antitumor action neurogenic cells of fetal rat. 3. Using primary cultures of rat brain gliomas to evaluate individual sensitivity of tumor cells to the studied biological product.

The purpose. Cultivation of mesenchymal stem cells (MSCs) from adipose tissue (AT) of experimental animals and humans to study induction of neurogenic differentiation. Materials and methods. AT samples from 30 experimental animals and 5 patients were used. MSCs cultures were studied for 5 passages. To induce neurogenic differentiation retinoic acid (1 mkM) and/or 5-azacytidine (1 mkM) were used. Cultures were observed in vivo, also according to cytological and immunohistochemical studies. Results. Fibroblast-like MSCs growth was observed in cell cultures from AT. At presence of specific differentiation factors they acquired neuron-like or astrocyte-like phenotype with positive staining to GFAP. Conclusions. In cultures of animal and human AT cells fraction of MSCs was derived, capable to induce neurogenic differentiating under the influence of retinoic acid and/or 5-azacytidine, as confirmed by immunohistochemistry.