Elaboration of new correction methods for microcirculatory disorders in the brain caused by persistent high blood pressure is important for both medicine and biology. We studied the influence of intracerebral transplantation of human mesenchymal stem cells (hMSCs) on the cerebral microcirculation in young (4 month) and mature (12 month) spontaneously hypertensive rats (SHRs). It was shown that hMSC transplantation increased the density of the microvascular network of young SHRs to 1.6 and 1.9 times the density of the arteriolar area of the microvascular network of the pia. The density of microvascular network of mature SHRs increased by 1.4–1.5 times after hMSC transplantation. The perfusion and tissue saturation of sensorimotor cortex of young SHRs restored to the level of young normotensive rats. In mature SHRs, the perfusion and tissue saturation of sensorimotor cortex did not increase. In conclusion, the intracerebral transplantation of hMSC almost completely restored microcirculation in the sensorimotor cortex of the brain of young SHRs and slightly improved microcirculation in mature SHRs.

Using a TV device for studying microcirculation (×40), we analyzed the density of the whole microvascular network and the density of arterioles in the pia mater of the sensorimotor cortex in SHR rats of different ages (3-4 and 12 months) after intracerebral transplantation of human mesenchymal stem cells. We found that the density of pial microvascular network in SHR rats receiving transplantation of human mesenchymal stem cells increased to a level observed in young Wistar-Kyoto rats.

We performed morphological analysis of the structure of rat hippocampus after ablation of the left sensorimotor cortex. Four experimental groups were formed: two control groups (intravenous and intracerebral injections of the culture medium) and two experimental groups (intravenous and intracerebral transplantation of MSC). Ten weeks after surgery, disturbed cytoarchitectonics and great number of dead neurons were found in all zones of the hippocampus in animals of the control groups. In animals receiving cell therapy, no pathological changes in the structure of the hippocampus were found: hyperchromatic neurons were absent and the cells had regular shape and closely adjoined to each other.

To assess safety and tolerability of treatment with autologic multipotent mesenchymal stem cells (MSC) in multiple sclerosis (MS), we have obtained autologic red bone marrow-derived MSC from 8 patients. Proliferation, immunophenotype and caryotype of MSC, their sterility, the absence of hemopoetic cells, chromosomal aberrations and signs of aging were controlled during the cell growth. The inverse injection of MSC in patient's blood was conducted in accordance to the elaborated protocol in a short intravenous infusion in dose 2.0 x 10(6)/kg of body mass once in 30 days. The duration of treatment was from 4 to 8 months. The efficacy of treatment was assessed after 4, 8 and 12 months. All patients tolerated repeated intravenous infusions of autologic MSC well with no significant side-effects as in the early as well in the remote periods of treatment. The distinct positive effect was seen in some cases 2 months after the beginning of treatment. The improvement of 0.5 point on EDSS was seen in 5/8 patients after 4 months. After 12 months, the improvement of 0.5-1 point on EDSS was seen in 6/8, stabilization in 1/8, progression in 1/8. These results revealed the safety of the elaborated protocol of treatment and the moderate clinical efficacy of treatment in non-curable patients or those with poor response to treatment that suggested continuing the study and enrollment of new patients.

We studied the effect of various methods of transplantation of mesenchymal stem cells on neuronal survival in rat brain 1 and 6 weeks after severe traumatic brain injury. It was found that intracerebral and systemic transplantation of mesenchymal stem cells improves neuronal survival in the piriform cortex of the contralateral hemisphere without affecting neuronal survival in the marginal zone of the traumatic cavity and amygdaloid nuclei. Intracerebral transplantation of mesenchymal stem cells increases the content of the astroglial component of the scar in the borderline zone of the traumatic cavity.

В настоящей работе представлены результаты лечения с помощью мезенхимных клеток (МСК) группы больных, страдающих тяжелой хронической ишемией нижних конечностей (ХИНК), сроки наблюдения за которыми составили от 2-х до 4-х лет. С ноября 2006 года по ноябрь 2010 года под нашим наблюдением находится 17 больных, страдающих ХИНК, которым проведено лечение с помощью МСК. Возраст больных от 36 до 76 лет, 62,5 года в среднем, женщины - 3, мужчин - 14. Атеросклеротическое поражение артерий нижних конечностей было у 15 человек, облитерирующий эндартериит у 2-х. У 6 человек (группа I) клинически имела место интенсивная перемежающаяся хромота (ПХ) - 25 метров и менее, иногда в сочетании с болями в покое (III стадия по Покровскому), у 11 больных (группа II) боли в покое в сочетании с разной тяжестью трофических изменений на стопе (IV стадия заболевания по Покровскому). Помимо клинической оценки ситуации у всех больных до и после введения МСК регистрировали лодыжечное давление (ЛД) на пораженной конечности; всем больным до введения МСК проводили ангиографию, у 5 человек это исследование повторено в сроки от 6 до 12 месяцев после введения клеток. МСК, полученные из костного мозга, вводили под местной анестезией в мышцы голени по задней ее поверхности в два ряда точек, всего 32-34 точки по 1,5 млн МСК на одну инъекцию. Состояние больных, местные изменения, контрольные измерения ЛД и т. п. оценивали и проводили до процедуры, в первые дни после имплантации клеток, а затем через 2-3 мес, 6 мес, 1, 1,5, 2 года и более. У 12 (70,5 %) из 17 больных с тяжелой ХИНК удалось сохранить пораженные ноги, получив, т.о. хороший результат от применения АМСК. Следует отметить, что у части этих пациентов клинически наблюдаемое улучшение не сопровождается повышением ЛД, не коррелирует с улучшением качеств путей коллатеральной компенсации на конечности и свидетельствует о неоднозначности механизмов, объясняющих положительный эффект клеточной терапии у обсуждаемых больных.

We studied the effects of intravenous and intracerebral transplantation of MSC on restoration of orientation and exploratory behavior of Wistar−Kyoto rats after removal of the left motor cortex. Removal of the motor cortex led to a significant reduction of the number of behavioral acts in the open field test. Two weeks after removal of the motor cortex and intravenous transplantation, the animals were as inhibited as the controls, but during the next 10 weeks, the behavioral status of these rats remained unchanged, while controls exhibited further behavioral degradation. After injection of MSC into the brain, the behavior of rats with trauma did not change in comparison with intact rats over 10 weeks.

The effects of the allogeneous demineralized bone transplants, seeded with bone marrow-derived multipotent mesenchymal stromal cells CMSCsJ, on the healing of the damaged articular cartilage and the subchondral bone were studied. It was shown that the use of allogeneous as well as autologous MSCs on the demineralized bone transplants and also on the transplants combined with type I collagen gel promote the remodeling of the regenerating tissue and the recovery of the histotypic cartilaginous and osseous structures in comparison with the articular surface regeneration without any therapy or after the procedure of the mosaic chondroplasty. Nevertheless, the positive effects of cell therapy were not clinically prominent, which means most probably in that case that the generally accepted experimental model is not adequate in full measure, but the approach used is promising.

The effects of the allogeneous demineralized bone transplants, seeded with bone marrow-derived multipotent mesenchymal stromal cells CMSCsJ, on the healing of the damaged articular cartilage and the subchondral bone were studied. It was shown that the use of allogeneous as well as autologous MSCs on the demineralized bone transplants and also on the transplants combined with type I collagen gel promote the remodeling of the regenerating tissue and the recovery of the histotypic cartilaginous and osseous structures in comparison with the articular surface regeneration without any therapy or after the procedure of the mosaic chondroplasty. Nevertheless, the positive effects of cell therapy were not clinically prominent, which means most probably in that case that the generally accepted experimental model is not adequate in full measure, but the approach used is promising.

Transplantation of human bone marrow mesenchymal stem cells is considered as a promising therapeutic approach to the therapy of many diseases. However, the problem of possible alterations of the properties of mesenchymal stem cells during their expansion in in vitro cultures before transplantation is not solved. In our study, one of two hundred examined cultures of mesenchymal stem cell cultures derived from donors without bone marrow pathologies and developed under standard culturing conditions demonstrated spontaneous disturbances in morphology, proliferation, and karyotype at early passages. The cells of this abnormal culture retained immunophenotype characteristic of normal mesenchymal stem cells, but some of them (15–25%) had numerous numerical and structural chromosome aberrations.