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Method for differentiating and identifying dry cells under action of exogenous neurotrophic factors

A technology of neurotrophic factors and stem cells, applied in the field of differentiation and identification of stem cells under the action of exogenous neurotrophic factors, can solve problems such as unsatisfactory recovery of spinal cord function, and achieve stable performance

Active Publication Date: 2014-04-02
北京泰圣康源生物医学研究院有限公司
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Problems solved by technology

However, the single application of one method cannot achieve satisfactory spinal cord function recovery, while the combined application of multiple methods has shown good spinal cord function recovery

Method used

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  • Method for differentiating and identifying dry cells under action of exogenous neurotrophic factors
  • Method for differentiating and identifying dry cells under action of exogenous neurotrophic factors

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Embodiment Construction

[0020] The present invention relates to a method for differentiation and identification of stem cells under the action of exogenous neurotrophic factors, and the specific terms are explained as follows:

[0021] DMEM is a medium containing various amino acids and glucose, which is developed on the basis of MEM medium. Compared with MEM, the amount of various ingredients has been increased, and it is divided into high-sugar type (less than 4500mg / L) and low-sugar type (less than 1000mg / L). The high-glucose type is conducive to the growth of cells anchored in one position, and is suitable for tumor cells that grow faster and are more difficult to attach.

[0022] HBSS: Hank's Balanced Salt Solution---Hank's Balanced Salt Solution

[0023] The formula of Hank's balanced salt solution (HBSS) is: 8 g / L NaCl, 0.4 g / L KCl, 1 g / L glucose, 60 mg / L KH2PO4, 47.5 mg / L Na2HPO4, NaHCO3, 0.35g / L, adjust pH to 7.2.

[0024] It is generally used to prepare medium or wash cells in the proces...

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Abstract

The invention relates to a method for differentiating and identifying dry cells under the action of exogenous neurotrophic factors. The method comprises the following steps: separating and amplifying spinal cord neural stem cells of an embryonic rat; culturing and differentiating neurosphere cells depending on EGF (epidermal growth factor) and FGF (fibroblast growth factor) under the action of different exogenous neurotrophic factors; constructing a recombinant adeno-associated virus containing HIF-1 (hypoxia-inducible factor-1) alpha and BDNF (brain-derived neurotrophic factor) sequences; performing in vitro transfection and analysis on the spinal cord neural stem cells of the rat, namely measuring an expression product and analyzing the bioactivity. The method is stable in performance.

Description

technical field [0001] The invention relates to a method for differentiation and identification of stem cells under the action of exogenous neurotrophic factors, belonging to the field of medical biotechnology. Background technique [0002] Spinal cord injury is the main cause of human paraplegia, and it has always been the focus and difficulty of neurosurgery research. The pathophysiological process of spinal cord injury includes three stages: primary spinal cord injury, secondary injury and spinal cord repair. Primary spinal cord injury is the death of nerve cells caused by physical damage at the time of injury. The mechanism of secondary spinal cord damage includes blood circulation disorders, changes in biochemical active substances, and energy metabolism disorders. The core problem is tissue ischemia and hypoxia. Loss of function after SCI is due to death of nerve cells, disruption of ascending and descending axons, and lack of efficient regeneration. Recent studies ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/861G01N33/68
Inventor 陈茂华
Owner 北京泰圣康源生物医学研究院有限公司
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