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

A technology of central nervous system and composition, which is applied in the direction of drug combination, nervous system diseases, microorganisms, etc., to achieve great reference value, convenient application, and reliable experimental results

Inactive Publication Date: 2007-10-03
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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.

[0034] (average)

[0035] 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 activ...

Embodiment 4

[0038]Example 4. Experimental transplantation of bone marrow MSCs-derived neural precursor cells and GDNF controlled-release biomaterials to treat monkey neurological dysfunction caused by artificial spinal cord injury. According to the methods described in Example 1 and Example 2, MSCs were isolated and cultured and GDNF controlled-release biomaterials were prepared. Animal models were prepared, and 8 healthy male rhesus monkeys weighing 4 kg were randomly divided into groups. Combined anesthesia with intramuscular ketamine hydrochloride injection and diazepam injection was used to make the thoracic spinal cord injury model according to the modified Allen's method.

[0039] The experimental design is as follows:

[0040] Transplantation group: on the 10th day after injury, digest and count the induced monkey MSCs, divide 3.0×10 6 200uL of PBS suspension made from cells / kg and controlled-release biomaterial containing 2ugGDNF, injected into the experimental group (n=4) at fi...

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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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/08A61K45/00A61P25/00
Inventor 邓宇斌
Owner SUN YAT SEN UNIV
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