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Transplantation of human neural cells for treatment of neurodegenerative conditions

A neural stem cell and symptom technology, applied in nervous system cells, animal cells, nervous system diseases, etc., can solve problems such as unmaintainable survival rate, regulatory and economic obstacles, etc.

Inactive Publication Date: 2007-12-05
NEURALSTEM INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This in turn requires a much larger scale of cell manufacturing, which raises further regulatory and economic hurdles
In addition, the viability of such cells in vivo cannot be maintained

Method used

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  • Transplantation of human neural cells for treatment of neurodegenerative conditions
  • Transplantation of human neural cells for treatment of neurodegenerative conditions
  • Transplantation of human neural cells for treatment of neurodegenerative conditions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0147] Example 1. Expansion of human spinal cord neural stem / progenitor cells

[0148] Spinal cords were obtained from at least one donor at a gestational age of approximately 7-8.5 weeks. Use mechanically developed Ca-free ++ and Mg ++ A single contiguous tissue of the spinal cord was dissociated in phosphate-buffered saline. The resulting cell suspension is then seeded into tissue culture plates precoated with poly-L-ornithine or poly-D-lysine and human fibronectin or other extracellular matrix proteins. Tissue culture treated plates or flasks were incubated with 100 μg / ml poly-D-lysine for 1 hour at room temperature. They were then washed three times with water and dried. They were then incubated with 25 mg / ml for 5 minutes at room temperature. Sometimes, 10 mg / ml fibronectin is used for 1 hour at room temperature. Sometimes, 1 mg / ml fibronectin was used for 18 hours at 37°C. Medium consisting of N2 (DMEM / F12 plus insulin, transferrin, selenium, putrescine and proges...

Embodiment 2

[0150] Example 2. Differentiation of Human Spinal Cord Neural Stem / Progenitor Cells

[0151] At any time during the expansion of NSCs, cultures can be differentiated by extracting mitogens, such as bFGF, from the culture. Differentiation of NSCs occurs approximately 1-3 days after removal of the mitogen and exhibits a distinct heterogeneous cell morphology. By approximately 4-7 days of differentiation, non-specific antigens such as MAP2c, tau, and type III β-tubulin can be visualized by immunostaining. By approximately 12-14 days, elongated fascicled axons and clear polarization of subcellular protein trafficking are evident throughout the culture. By approximately day 28, synaptic proteins, such as synapsin and synaptophysin, are localized to axon terminals and appear as punctate staining. Another feeder layer of astrocytes can be provided to further promote long-term maturation of neurons. As shown in Figure 3, differentiation of human spinal NSCs produces mixed cultures ...

Embodiment 3

[0152] Example 3. Expansion of human midbrain neural stem / progenitor cells

[0153] Obtain one midbrain tissue at a fetal age of 7-8.5 weeks. NSCs from midbrain tissue were obtained as described in Example 1. Cells were serially passaged through a 160-day pure culture period, and the resulting expansion is shown in FIG. 4 . Throughout the expansion period, NSCs stably maintained their pluripotent and neurogenic potential as well as their differentiation potential to give rise to dopaminergic neurons. Dopaminergic neurons were assessed by neuronal expression of tyrosine hydroxylase (TH) and dopamine transporter (DAT).

[0154] DAT expression is a hallmark of dopaminergic neurons. DAT expression in neurons can be assessed by measuring its effect in transporting radioisotope-labeled dopamine across the synaptic membrane of differentiated neurons in culture. The effect of DAT in cultures from differentiated human midbrain NSCs and monoclonal derived human midbrain NSCs was asses...

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Abstract

The invention relates generally to methods of treating spasticity, rigidity, or muscular hyperactivity conditions by introducing a portion of an expanded population of neural stem cells into an area of a recipient spinal cord.

Description

technical field [0001] The methods disclosed herein relate to methods of treating disorders by transplantation of cells that are particularly beneficial for such methods of treatment. In particular, the disclosed methods provide methods for treating neurodegenerative disorders with neural stem cells (NSCs). Background technique [0002] Neurodegenerative disorders are characterized by symptoms accompanied by neuronal degeneration caused by disease, genetic conditions, or trauma (eg, traumatic or ischemic spinal cord or brain injury). [0003] Spinal circuits that control contraction of limb skeletal muscles include excitatory motor neurons and inhibitory GABAergic (ie, GABA-producing) and glycinergic (ie, glycine-producing) interneurons. Motor neurons are nerves that originate in the anterior horn of the gray matter of the spinal cord. Axons of motor neurons emerge from spinal cord segments as efferent motor fibers that stimulate muscle fibers. Pulses made by motor neuron...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00A01N63/00C12N5/0797
CPCC12N2501/115C12N2501/11A61K35/30C12N2501/148C12N5/0623A61P21/00A61P21/02A61P25/00A61P25/02A61P25/04A61P25/08A61P25/14A61P25/28A61P29/00A61P9/00A61P9/10A61K9/0085C12N2501/113
Inventor M·马莎拉K·K·乔T·G·黑兹尔垣花修V·科利亚索斯阎军P·J·赖尔M·J·维拉多
Owner NEURALSTEM INC
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