RESUMEN
This paper shows for the first time that co-transplantation of human olfactory ensheathing cells with neurotrophin-3 into spinal cord cysts is more effective for activation of remyelination than transplantation of cells with brain-derived neurotrophic factor and a combination of these two factors. The studied neurotrophic factors do not affect proliferation and migration of ensheathing cells in vitro. It can be concluded that the maximum improvement of motor function in rats receiving ensheathing cells with neurotrophin-3 is largely determined by activation of remyelination.
Asunto(s)
Factor Neurotrófico Derivado del Encéfalo , Neurotrofina 3 , Bulbo Olfatorio , Remielinización , Animales , Ratas , Neurotrofina 3/metabolismo , Humanos , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Factor Neurotrófico Derivado del Encéfalo/farmacología , Remielinización/fisiología , Bulbo Olfatorio/citología , Proliferación Celular , Médula Espinal/metabolismo , Vaina de Mielina/metabolismo , Vaina de Mielina/fisiología , Células Cultivadas , Movimiento Celular , Quistes/patología , Femenino , Quistes del Sistema Nervioso Central/cirugía , Quistes del Sistema Nervioso Central/patologíaRESUMEN
We analyzed the main approaches to the modeling of Alzheimer's disease for studying the effectiveness of cell therapy. Recent advances in regenerative medicine in the field of neuroscience create prospects for the use of various cell preparations for the treatment of Alzheimer's disease. Experimental data on the use of neural stem/progenitor cells, mesenchymal stem cells, embryonic stem cells, and induced pluripotent stem cells in various models of Alzheimer's disease are presented. Of particular importance is the standardization of protocols. The use of a standardized protocol in modeling of Alzheimer's disease will allow a comparative analysis of the effectiveness and safety of treatment to identify the optimal cell preparation. The data obtained on experimental animals can form the basis for further preclinical and clinical studies of cell therapy for Alzheimer's disease.
RESUMEN
Alzheimer's disease was modeled in female Wistar rats aged 4 months by stereotaxic bilateral injection of a synthetic peptide ß-amyloid (Aß1-42) into the hippocampus. Behavioral tests (open field, Y-maze, passive avoidance, and Morris water maze) revealed significant impairment of memory and spatial navigation 8 weeks after ß-amyloid administration. At this term, the cognitive impairments typical of Alzheimer's disease are reproduced. The experimental model of Alzheimer's disease proposed by us can be used in preclinical studies of drugs for the treatment of this pathology.
Asunto(s)
Enfermedad de Alzheimer , Ratas , Femenino , Animales , Enfermedad de Alzheimer/patología , Ratas Wistar , Escala de Evaluación de la Conducta , Aprendizaje por Laberinto , Péptidos beta-Amiloides/metabolismo , Hipocampo/metabolismo , Modelos Teóricos , Modelos Animales de Enfermedad , Fragmentos de Péptidos/uso terapéutico , Trastornos de la Memoria/tratamiento farmacológicoRESUMEN
We developed a viral vector Ad5/35-CAG-mBDNF expressing the mature form of BDNF (mBDNF). On the basis of olfactory ensheathing cells transduced with this adenovector, a new gene-cell construct was obtained. In experiments in vitro, high viability of the transduced olfactory ensheathing cells and enhanced secretion of BDNF by these cells were observed. It is possible that a new gene-cell construct will significantly increase the regenerative effects of transplanted olfactory ensheathing cells.
Asunto(s)
Mucosa Olfatoria , Traumatismos de la Médula Espinal , Factor Neurotrófico Derivado del Encéfalo , Vectores Genéticos/genética , Humanos , Regeneración Nerviosa/genética , Bulbo Olfatorio , Médula Espinal , Traumatismos de la Médula Espinal/genética , Traumatismos de la Médula Espinal/terapiaRESUMEN
Neurotrophin-3 enhances the effectiveness of human olfactory ensheathing cells in improving hind limb mobility in rats with post-traumatic cysts of the spinal cord. Transplantation of olfactory ensheathing cells into spinal cord cysts reduced their size; neurotrophin-3 did not modulate this effect. Combined preparation of human olfactory ensheathing cells and neurotrophin- 3 can be used in neurosurgery for the treatment of patients with spinal cord injuries.
Asunto(s)
Tratamiento Basado en Trasplante de Células y Tejidos , Quistes , Neurotrofina 3 , Traumatismos de la Médula Espinal , Animales , Trasplante de Células , Quistes/terapia , Humanos , Factores de Crecimiento Nervioso/genética , Regeneración Nerviosa , Neurotrofina 3/farmacología , Ratas , Médula Espinal , Traumatismos de la Médula Espinal/terapiaRESUMEN
A gene-cell construct based on rat olfactory mucosa ensheathing cells transduced with an adenoviral vector encoding a mature form of brain neurotrophic factor (mBDNF) was transplanted into post-traumatic cysts of rat spinal cord. Transplantation of the gene-cell construct improved motor activity of the hind limbs and reduced the size of cysts in some animals. However, comparison of the effects of transduced and non-transduced ensheathing cells revealed no significant differences. In parallel in vitro experiments, a decrease in the proliferation of transduced cells compared to non-transduced cells was observed. It is likely that mBDNF reduces proliferation of transduced cells, which can affect their efficiency. The therapeutic efficacy of the new gene-cell construct is most likely provided by the cellular component.
Asunto(s)
Quistes , Traumatismos de la Médula Espinal , Animales , Factor Neurotrófico Derivado del Encéfalo/genética , Quistes/genética , Quistes/terapia , Regeneración Nerviosa , Mucosa Olfatoria , Ratas , Recuperación de la Función , Médula Espinal , Traumatismos de la Médula Espinal/genética , Traumatismos de la Médula Espinal/terapiaRESUMEN
We studied the effect of transplantation of ensheathing cells obtained from the olfactory mucosa of rats and humans on the size of posttraumatic spinal cord cysts. MRI examination showed that transplantation of these cells into experimental posttraumatic cysts of the spinal cord led to a significant decrease in cyst volume and even their complete disappearance in two animals receiving transplantation of rat or human cells. These findings attested to regenerative processes developing as a result of ensheathing cell transplantation. Further studies in this field will be aimed at elucidation of the mechanisms underlying spinal cord regeneration in the area of posttraumatic cysts after transplantation of ensheathing cells.
Asunto(s)
Quistes , Traumatismos de la Médula Espinal , Animales , Trasplante de Células , Regeneración Nerviosa , Bulbo Olfatorio , Mucosa Olfatoria , Ratas , Médula Espinal , Traumatismos de la Médula Espinal/terapiaRESUMEN
In this work, an optimal protocol was developed for obtaining adhesion culture of neural stem/progenitor cells (NSPC) of rat olfactory mucosa. During the development of the protocol, the conditions for cell culturing on adhesion substrates fibronectin and laminin in DMEM/F-12 and neurobasal media with the same culture additives were compared. Cell proliferation was maximum during culturing on both substrates in the neurobasal medium. Using the immunofluorescence method, we found that culturing on fibronectin in the neurobasal medium ensured maximum (52.22%) content of nestin-positive cells in comparison with other culturing conditions. The highest percentage of ßIII-tubulin-positive cells was detected in cultures growing on fibronectin in the neurobasal medium and in DMEM/F-12 (79.11 and 83.52%, respectively). Culturing in adhesion cultures in the neurobasal medium on fibronectin allowed obtaining cultures enriched with NSPC and neurons differentiating from them in a quantity sufficient for further transplantation. The developed protocol can be recommended for obtaining NPSC from human olfactory mucosa for the treatment of spinal cord injuries.
Asunto(s)
Tratamiento Basado en Trasplante de Células y Tejidos/métodos , Medios de Cultivo/farmacología , Células-Madre Neurales/citología , Neuronas/citología , Mucosa Olfatoria/citología , Animales , Biomarcadores/metabolismo , Adhesión Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Medios de Cultivo/química , Fibronectinas/farmacología , Expresión Génica/efectos de los fármacos , Humanos , Laminina/farmacología , Nestina/genética , Nestina/metabolismo , Células-Madre Neurales/efectos de los fármacos , Células-Madre Neurales/metabolismo , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Mucosa Olfatoria/efectos de los fármacos , Mucosa Olfatoria/metabolismo , Cultivo Primario de Células , Ratas , Ratas Wistar , Traumatismos de la Médula Espinal/terapia , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismoRESUMEN
In experiments on rats, co-transplantation of olfactory ensheathing cells of the human olfactory mucosa and neural stem/progenitor cells from the same source into post-traumatic cysts of the spinal cord led to improvement of the motor activity of the hind limbs and reduced the size of the cysts in some animals by 4-12%. The transplantation of a combination of the olfactory mucosa cells is effective and can be used in preclinical trials for the treatment of spinal cord injuries.
