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Bone marrow interstital stem cell preparation and its combined use with controlled release neurotrophic factor

A neurotrophic factor, stem cell technology, applied in tissue culture, biochemical equipment and methods, microorganisms, etc., to achieve comprehensive observation indicators, significant reference value, and reliable experimental results.

Inactive Publication Date: 2005-12-21
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still doubts about the activity of neurons after MSC-induced differentiation, whether transplanted into the central nervous system can continue to survive, and whether they can promote the recovery of neurological function.

Method used

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  • Bone marrow interstital stem cell preparation and its combined use with controlled release neurotrophic factor
  • Bone marrow interstital stem cell preparation and its combined use with controlled release neurotrophic factor
  • Bone marrow interstital stem cell preparation and its combined use with controlled release neurotrophic factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Example of preparation technology of improved bone marrow mesenchymal stem cells (MSCs). Bone marrow of healthy monkey ilium was extracted, mononuclear cells were isolated, inoculated in L-DMEM culture medium containing 15% FBS (bFGF, 3 ng / ml was added therein), and cultured in plastic culture flasks. The 5th to 8th generation bone marrow MSCs were taken for identification. The methods included flow cytometry to detect their surface antigen markers, and in vitro induction to observe their osteogenic and adipogenic multidirectional differentiation potential. 48 hours before transplantation, Brdu was labeled at a concentration of 10umol / L and counted for later use.

Embodiment 2

[0034] Example 2. Determination of the optimal induction time of MSCs for transplantation in vitro. The MSCs-induced cells were divided into 0 hour group (control), 1 hour group, 1.5 hour group, 2 hour group and 3 hour group, each group was divided into 6 samples, and nestin (NESTIN), neuron-specific enzymatic Alcoholase (NSE) immunohistochemical staining, and flow cytometry for apoptosis detection.

[0035] (Average) NESTIN Positive Rate (%) NSE Positive Rate (%) Apoptotic Percentage (%)

[0036] 0 hour group 0 0 2.7

[0037]1 hour group 10 15 2.8

[0038] 1.5 hour group 25 30 3.2

[0039] 2 hours group 10 35 5.7

[0040] 3 hours group 3 55 7.2

[0041] It can be seen from the results that 1.5 hours of induction can not only ensure the effect of induction but also ensure the activity of cells, which is the best induction time. At present, the transplantation of induced cells does not pay attention to the activity of transplanted cells. If the activity of transplanted ce...

Embodiment 3

[0042] Example 3. Determination of the optimal ratio of transplantation-induced MSCs and controlled-release neurotrophic factors. The controlled-release GDNF with different GDNF content was placed in the culture medium for 1 week, and then the culture medium was made into a nerve induction solution for 3.0×10 6 Monkey MSCs were induced, 6 samples per group, and the percentage of apoptosis was detected for 3 hours.

[0043] GDNF content ug Apoptotic percentage (average)

[0044] 0 7.2

[0045] 0.25 6.4

[0046] 0.375 5.8

[0047] 0.5 3.8

[0048] 0.625 3.8

[0049] 0.75 3.6

[0050] It can be seen from the results that when the GDNF content is 0.5, the apoptosis percentage of induced cells can be significantly reduced, and the effect of increasing the content has no obvious increase. Therefore, 3.0×10 6 Controlled-release GDNF with a GDNF content of 0.5ug is the best ratio, because the total amount of transplanted cells in this experiment is 1.2×10 7 , the total amount ...

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Abstract

The invention relates to preparation of mass stem cell among the bone marrow and the combined use with controlled-release nervous nutrition factor. The mass stem cell MSC preparation method includes the cultivation of mass stem cell among the bone marrow; the external inducing of the MSC; combied use of the transplanting and inducing MSC among bone marrow and controlling the release of nervous nutrition factor GDNF. The invention is the effective tactics of treating damaging and relevant disease of spinal cord, have wide clinical development prospects.

Description

[technical field] [0001] The present invention relates to a method for the combined application of stem cell preparations and controlled-release neurotrophic factors for the treatment of nervous system injuries, in particular, to the preparation of bone marrow mesenchymal stem cell preparations and the optimal in vitro neural induction time, and its combination with controlled-release neurotrophic factors The optimal mixing ratio of factors, a new technology for the treatment of brain and spinal cord central nervous system and other diseases. [technical background] [0002] For the paralysis and other nervous system dysfunction caused by nervous system damage, there is often no effective clinical treatment. Some people use nervous system cell transplantation to promote the functional recovery of nerve damage, but the donor nervous system cells are mainly derived from fetal brain or other accidents. For those who died, there were immune rejection problems due to insufficient ...

Claims

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Application Information

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IPC IPC(8): C12N5/08
Inventor 邓宇斌
Owner SUN YAT SEN UNIV
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