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Method for preparing bone marrow-derived neural stem cell for expressing anti-inflammatory cell factor IL-10

An anti-inflammatory cytokine and neural stem cell technology, applied in the field of preparation of bone marrow-derived neural stem cells expressing anti-inflammatory cytokine IL-10, can solve the problems of unsatisfactory therapeutic effect, weak anti-inflammatory effect, and inability to relieve damage

Inactive Publication Date: 2010-12-15
LIAONING UNIV OF TRADITIONAL CHINESE MEDICINE
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Problems solved by technology

However, due to its weak anti-inflammatory effect, it cannot relieve the damage to the myelin sheath and neurons caused by the persistent inflammation of the central nervous system, so the therapeutic effect is not very ideal.

Method used

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  • Method for preparing bone marrow-derived neural stem cell for expressing anti-inflammatory cell factor IL-10
  • Method for preparing bone marrow-derived neural stem cell for expressing anti-inflammatory cell factor IL-10
  • Method for preparing bone marrow-derived neural stem cell for expressing anti-inflammatory cell factor IL-10

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Embodiment Construction

[0031] A method for preparing bone marrow-derived neural stem cells expressing anti-inflammatory cytokine IL-10, comprising the steps of:

[0032] 1. Generation and identification of mouse bone marrow-derived neural stem cells:

[0033] 1) Generation and culture of bone marrow-derived neural stem cells (BM-NSCs): Take the fibula of 8-12 week old C57BL / 6 mice, extract whole bone marrow cells, and use microbead-labeled antibodies from Miltenyi Biotec, Germany and magnetic cell sorter (QuadroMACS TM Separator) screened mesenchymal stem cells in two steps. First, the Lineage Cell Depletion Kit was used to remove mature hematopoietic cells, such as T cells, B cells, monocytes / macrophages, granulocytes and red blood cells, and committed progenitor cells (CD5 + , CD45 + , CD11b + , Gr-1 + , TER119 + , 7 / 4 + ), reserved allonegative cells (lineage - ); then use anti-CD117 microbeads (CD117(c-kit) Microbeads) to screen out the lineage - / CD117(c-kit) + Mesenchymal stem cells...

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Abstract

The invention provides a method for preparing a novel neural stem cell which is derived from bone marrow and can express an anti-inflammatory cell factor IL-10. The method comprises the following steps of: (1) generating and cultivating a bone marrow-derived neural stem cell and cultivating a brain-derived neural stem cell in vitro; and (2) updating the bone marrow-derived neural stem cell and the brain-derived neural stem cell by self and comparing the differentiation capacities to verify that the two cells have the same self update and differentiation capacity and both can be differentiated into neuron and neuroglia cell energy. The neural stem cell prepared by the method can be used for treating MS (Multiple Sclerosis) diseases and other various nerve system diseases, provides a brand new approach for the MS and other nerve system diseases and has wide application prospect.

Description

Technical field: [0001] The present invention relates to a method for establishing a new neural stem cell line derived from bone marrow that can express the anti-inflammatory cytokine IL-10, and the therapeutic effect of the new neural stem cell that combines cell replacement and gene therapy on multiple sclerosis . Background technique: [0002] Neural stem cells (NSCs) refer to a group of cell populations that have self-renewal ability and can differentiate into neurons and glial cells (oligodendrocytes, astrocytes). In recent years, many scholars have used neural stem cell transplantation to treat multiple sclerosis (MS), Parkinson's disease, Alzheimer's disease and other nervous system diseases and nerve injuries, and achieved remarkable therapeutic effects. However, the neural stem cells originate from embryos or adult animals around the lateral ventricle and hippocampus, which are difficult to obtain and have great damage. The neural stem cells derived from embryos ca...

Claims

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Application Information

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IPC IPC(8): C12N5/0775C12N5/0797C12N15/63C12N5/10
Inventor 杨静娴王东孟宪生康廷国徐志立
Owner LIAONING UNIV OF TRADITIONAL CHINESE MEDICINE
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