Extracting method and special culture medium for rat omentum majus adipose source mesenchymal stem cells

A technology of adipose-derived stem cells, applied in animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve the problems of low immunogenicity and low immunogenicity

Inactive Publication Date: 2013-07-03
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

(2) Low immunogenicity: when adipose-derived mesenchymal stem cells are used for stem cell transplantation, due to the high content of autologous adipose tissue, they are mostly used for autologous transplantation, so they have low immunogenicity

Method used

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  • Extracting method and special culture medium for rat omentum majus adipose source mesenchymal stem cells
  • Extracting method and special culture medium for rat omentum majus adipose source mesenchymal stem cells
  • Extracting method and special culture medium for rat omentum majus adipose source mesenchymal stem cells

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Experimental program
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Embodiment 1

[0040] Rat omentum extracts adipose-derived mesenchymal stem cells and uses a special medium, including the following steps

[0041] Preparation of special medium for rat omentum adipose-derived mesenchymal stem cells: Add 25ml of fetal bovine serum, 5ml of antibiotics (penicillin and streptomycin), 5mg of insulin, Rat recombinant fibroblast growth factor 2.5ug, rat epidermal growth factor 2.5ug, thyronine 5ug, sterilized by 0.22μm filter, stored at 4°C for later use.

[0042] Extraction and culture steps of adipose-derived mesenchymal stem cells from rat omentum:

[0043] 1. Take a healthy rat with a weight of 100g-130g, place it on a sterile operating table after anesthesia, cut it open from the side of the rat’s abdomen with a sterile scalpel, take about 3g of omentum fat, and put it in a pre-cooled in PBS, suture the wound. Then wash with PBS several times to wash away the residual blood, cut into 1mm 3 left and right small pieces;

[0044] 2. Add PBS to wash, and cent...

Embodiment 2

[0050] Example 2: Identification of surface antigens of omentum adipose-derived mesenchymal stem cells of three generations of rats.

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Abstract

The invention discloses a separating and extracting method and special culture medium for rat omentum majus adipose mesenchymal stem cells. The method comprises the following steps: taking omentum majus fat of a healthy Sprague Dawley (SD) rat under an aseptic condition; cutting into pieces, centrifuging after being mixed with cold phosphate buffer to wipe off partial adipose tissues; digesting and centrifuging by mixed collagenase of I type collagenase and II type collagenase to wipe off upper adipose tissues; centrifuging again; suspending again in the special culture medium containing particular chemical substances and growth factor; moving into a culture flask; putting into an incubator to subculture continuously. The method is used to extract the omentum majus adipose source mesenchymal stem cells from a rat body; meanwhile, during the culturing, the problem that cell senescence and cell differentiation easily occur when stem cells are cultured in vitro can be solved through the special culture medium containing particular chemical substances and growth factor; and the omentum majus adipose source mesenchymal stem cells that can maintain characteristics of stem cells for a long time can be obtained.

Description

technical field [0001] The invention relates to a method for extracting rat omentum fat-derived mesenchymal stem cells and a special culture medium. Background technique [0002] Mesenchymal stem cells (MSCs) are pluripotent stem cells derived from mesoderm and ectoderm, and are classified as adult stem cells according to the developmental process. Mesenchymal stem cells have the same self-renewal ability as embryonic stem cells, and have the potential to differentiate into multiple germ layers. Under specific induction conditions, they can differentiate into bone, cartilage, fat, nerve, liver cells and other tissue cells. Mesenchymal stem cells exist in a variety of tissues, and mesenchymal stem cells have been successfully obtained from bone marrow, fat, muscle and other tissues. In 2001, Zuk extracted a kind of mesenchymal stem cells with self-renewal and differentiation from the fat fluid in patients undergoing liposuction, and this kind of stem cells were named as adip...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 胡飞虎孙博肖忠党
Owner SOUTHEAST UNIV
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