Extract of egg cell from human or animal and use for amplifying stem cell as well as induction and differentiation

An egg cell and extract technology, which is applied to human or animal egg cell extract and its application in stem cell expansion and induced differentiation, can solve problems such as different reagents, and achieve good repeatability and stability.

Inactive Publication Date: 2005-01-12
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 reported methods and reagents are different, and there is no standard

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, the preparation of egg cell extract

[0044] (1) Take 10 grams of human or animal ovum cells, wash them with normal saline for 2-3 times, and add an equal amount of normal saline for subsequent use;

[0045] (2) Under the condition of 4°C, tissue homogenizer or masher is homogenized 3-5 times at 10,000-30,000 rpm, 1 minute each time;

[0046] (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 no higher than 42°C. Repeat the freezing and thawing for 3-5 times under the above conditions. According to conventional smear microscope observation, it was proved that the cells were completely broken;

[0047] (4) Settling naturally for 2-10 hours under the condition of 1-4°C, absorbing the supernatant;

[0048] (5) Filter with 3-4 layers of sterile gauze to remove sediment and protein flocs;

[0049] (6) Centrifuge at 400-6008 for 30 minutes at 4°C, and take t...

Embodiment 2

[0052] Example 2, Mesenchymal and Cell Expansion

[0053] Kit, egg cell extract protein 10mg / ml

[0054] The dosage of each component per 100ml kit:

[0055] (1) 90 ml mixed medium of DMEM and F12;

[0056] (2) Human egg cell extract 10ml;

[0057] Each component is individually packaged.

[0058] The kit may contain conventionally used effective doses of cell growth-promoting factors, antioxidants, antibiotics, and the like.

[0059] Amplification method:

[0060] 1. Isolation and purification of mesenchymal stem cells: mononuclear cells are isolated from bone marrow, fat and other tissues according to conventional methods, and the primary adherent culture of the cells is carried out first, and then positive and negative immunization is carried out with the specific marker antigen on the surface of mesenchymal stem cells Screening to obtain pure mesenchymal stem cells. If the number of cells is sufficient, the isolated mononuclear cells can also be directly used for sep...

Embodiment 3

[0063] Example 3, induced differentiation of mesenchymal stem cells into nerve cells

[0064] Kit, egg cell extract protein 10mg / ml nerve cell extract protein 10mg / ml

[0065] The dosage of each component per 100ml kit:

[0066] (1) 80 ml mixed medium of DMEM and F12;

[0067] (2) Human egg cell extract 10ml;

[0068] (3) 10 ml of nerve cell extract;

[0069] Each component is individually packaged.

[0070] The kit may contain conventionally used effective doses of cell growth-promoting factors, antioxidants, antibiotics, and the like.

[0071] Induction culture method:

[0072] 1. Isolation and purification of mesenchymal stem cells: mononuclear cells are isolated from bone marrow, fat and other tissues according to conventional methods, and the primary adherent culture of the cells is carried out first, and then positive and negative immunization is carried out with the specific marker antigen on the surface of mesenchymal stem cells Screening to obtain pure mesenchym...

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PUM

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Abstract

An extracted material from ovum of human being or animal is a new reagent for stem cells cloning and inducing technique. The preparing process of the material includes collecting raw material of ovum, washing, homogenating, freezing and melting, precipitating, picking top clear solution out, holding for impurity removal and loading.

Description

field of invention [0001] The invention relates to a biological cell extract and application technology, in particular to a human or animal egg cell extract and its application in stem cell expansion and induced differentiation. Background technique [0002] For many years, the research on egg cells has mostly involved in the field of fertility technology, and the research on egg cell extracts and their related technologies and applications in stem cell expansion and induced differentiation has not been reported. [0003] Since 1997, stem cell research has made important progress. Studies have found that human mesenchymal stem cells can be induced to differentiate into mature cells of nerve, muscle, skin, cartilage, fat, tendon, liver, blood and tissue types. After amplification and induction, it can be well integrated with corresponding tissue cells and perform specific biological functions. It can be used for transplantation to treat various tissue degeneration, tissue cel...

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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