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Neurocyte extract and its uses in mesenchyma stem cell differentiation induction

A technology of nerve cells and extracts, applied in biochemical equipment and methods, microorganisms, tissue culture, etc., can solve the problems of unstable application effect and non-formed standards, and achieve a good effect of inducing differentiation rate

Inactive Publication Date: 2005-01-26
成都军区昆明总医院 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research methods and inducing reagents in the prior art are different, the application effect is unstable, and no standard has been formed

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, the preparation of nerve cell extract

[0033] (1) Take human or animal sciatic nerve cell tissue, immediately inject normal saline through the artery and vein, remove the red blood cells remaining in the blood vessel to the greatest extent, shave off unnecessary components such as the capsule, connective tissue, and blood vessels, cut them into pieces, and add an equal amount of physiological salt water for use;

[0034] (2) Under the condition of 4°C for tissue homogenizer or mashing machine, 10,000-30,000 rpm for 3-5 times, 1 minute each time; it should be noted that the homogenizer should not be overheated, otherwise part of the tissue fluid will lose its activity;

[0035] (3) Store the homogenate in a -20°C refrigerator until it is completely frozen, generally for more than 24 hours, and then completely thaw it in water at a temperature not higher than 42°C. Repeat the freezing and thawing for 3-5 times under the above conditions. Observation with a...

Embodiment 2

[0039] Example 2, the application of MSC induced differentiation into nerve cells

[0040] The application of the kit, egg cell extract aqueous solution, containing 10 mg / ml protein, the above-mentioned nerve cell extract aqueous solution containing 10 mg / ml protein, prepared per 100 ml kit, each component is individually packaged. Dosage:

[0041] (1) 80 ml mixed medium of DMEM and F12

[0042] (2) Human egg cell extract liquid 10ml

[0043] (3) Nerve cell extract water 10ml

[0044] The kit may contain conventionally used effective doses of cell growth-promoting factors, antioxidants, antibiotics, and the like.

[0045] Induction culture method:

[0046] 1. Isolation and purification of mesenchymal stem cells: mononuclear cells are isolated from bone marrow, adipose and other tissues according to conventional methods, and the cells are first adhered to the wall for primary culture and then positively and negatively immunized with surface-specific marker antigens of mesenchy...

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PUM

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Abstract

The invention relates to the human being or animal nerve cell extracts and the application thereof for inducing and differentiating the mesenchyme to nerve cell in vitro, which provides a new induction reagent for mesenchym induction technology. The preparation steps are: removing blood cell, connective tissue and capsule from nerve cell tissue, conducting cell homogenate, cell disrupting, precipitating, taking supernatant, separating and removing substance with the molecular weight over 200 thousand Da, subpackaging. It can be applied both in the organic combination of nerve cell extractive and the prior part and in the preparation of reagent box. The invention in the application for inducing and differentiating the mesenchym into nerve cell shows a favorable effect, with the induction differentiation rate being over 90%. The inducing differentiated nerve cell can be used for transplantation to treat multiple nerve diseases, having a favorable application prospect in clinical practice

Description

field of invention [0001] The invention relates to a biological cell extract and its application technology, in particular to a human or animal nerve cell extract and its application technology in inducing differentiation of mesenchymal stem cells into nerve cells. Background technique [0002] Since 1997, stem cell research has made important progress. It has been found that human mesenchymal stem cells (MSCs) can be induced to differentiate into mature cells of nerve, muscle, skin, cartilage, fat, tendon, liver, blood and tissue types. After transplantation into the body, it is well integrated with the corresponding tissue cells, exerts specific biological functions, and can be used for transplantation to treat various diseases. The above research results suggest that neural-like cells derived from MSC expansion and differentiation can be used for transplantation to treat neurological diseases such as Alzheimer's disease, Parkinson's disease, brain atrophy, and peripheral ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/08
Inventor 潘兴华庞荣清李俊陆家海
Owner 成都军区昆明总医院
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