ABSTRACT
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.
Subject(s)
Brain-Derived Neurotrophic Factor , Neurotrophin 3 , Olfactory Bulb , Remyelination , Animals , Rats , Neurotrophin 3/metabolism , Humans , Brain-Derived Neurotrophic Factor/metabolism , Brain-Derived Neurotrophic Factor/pharmacology , Remyelination/physiology , Olfactory Bulb/cytology , Cell Proliferation , Spinal Cord/metabolism , Myelin Sheath/metabolism , Myelin Sheath/physiology , Cells, Cultured , Cell Movement , Cysts/pathology , Female , Central Nervous System Cysts/surgery , Central Nervous System Cysts/pathologyABSTRACT
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.
ABSTRACT
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.
Subject(s)
Alzheimer Disease , Rats , Female , Animals , Alzheimer Disease/pathology , Rats, Wistar , Behavior Rating Scale , Maze Learning , Amyloid beta-Peptides/metabolism , Hippocampus/metabolism , Models, Theoretical , Disease Models, Animal , Peptide Fragments/therapeutic use , Memory Disorders/drug therapyABSTRACT
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.
Subject(s)
Olfactory Mucosa , Spinal Cord Injuries , Brain-Derived Neurotrophic Factor , Genetic Vectors/genetics , Humans , Nerve Regeneration/genetics , Olfactory Bulb , Spinal Cord , Spinal Cord Injuries/genetics , Spinal Cord Injuries/therapyABSTRACT
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.
Subject(s)
Cell- and Tissue-Based Therapy , Cysts , Neurotrophin 3 , Spinal Cord Injuries , Animals , Cell Transplantation , Cysts/therapy , Humans , Nerve Growth Factors/genetics , Nerve Regeneration , Neurotrophin 3/pharmacology , Rats , Spinal Cord , Spinal Cord Injuries/therapyABSTRACT
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.
Subject(s)
Cysts , Spinal Cord Injuries , Animals , Brain-Derived Neurotrophic Factor/genetics , Cysts/genetics , Cysts/therapy , Nerve Regeneration , Olfactory Mucosa , Rats , Recovery of Function , Spinal Cord , Spinal Cord Injuries/genetics , Spinal Cord Injuries/therapyABSTRACT
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.
Subject(s)
Cysts , Spinal Cord Injuries , Animals , Cell Transplantation , Nerve Regeneration , Olfactory Bulb , Olfactory Mucosa , Rats , Spinal Cord , Spinal Cord Injuries/therapyABSTRACT
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.
Subject(s)
Cell- and Tissue-Based Therapy/methods , Culture Media/pharmacology , Neural Stem Cells/cytology , Neurons/cytology , Olfactory Mucosa/cytology , Animals , Biomarkers/metabolism , Cell Adhesion/drug effects , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Culture Media/chemistry , Fibronectins/pharmacology , Gene Expression/drug effects , Humans , Laminin/pharmacology , Nestin/genetics , Nestin/metabolism , Neural Stem Cells/drug effects , Neural Stem Cells/metabolism , Neurons/drug effects , Neurons/metabolism , Olfactory Mucosa/drug effects , Olfactory Mucosa/metabolism , Primary Cell Culture , Rats , Rats, Wistar , Spinal Cord Injuries/therapy , Tubulin/genetics , Tubulin/metabolismABSTRACT
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.
