Mesenchymal stem cells from canine fetal membranes, preparation method and used culture medium
A technology of mesenchymal stem cells and culture medium, applied in the field of culture medium used by mesenchymal stem cells in the treatment of canine diseases, can solve problems such as being unsuitable for canine animals
Examples
Embodiment 1
[0044] Example 1. Preparation of mesenchymal stem cells from canine fetal membranes
[0045] (1) Disinfection and cleaning: In the biological safety cabinet, take out the placenta from the placenta sample collection box, put it in a stainless steel cup, and use alcohol (75% alcohol) and normal saline to rinse the surface repeatedly (alcohol soaking disinfection time 30s)) , disinfect the placenta; then use surgical forceps to slowly tear off the fetal membranes, and place them in a glass dish. After removing the blood vessels, use phosphate buffer (0.025M sodium dihydrogen phosphate buffer at pH 6.5) to rinse again , remove surface dirt and impurities;
[0046](2) Digestion treatment: in another cell culture plate, the fetal membrane tissue obtained in step (1) is cut into tissue blocks (cubic blocks of about 0.1 cubic centimeters), and the tissue blocks are put into digestive enzyme solution (preparation method : Add 0.1g of type I collagenase to 100ml of DMEM-F12, then fil...
Embodiment 2
[0050] Example 2. Preparation of mesenchymal stem cells from canine fetal membranes
[0051] (1) Disinfection and cleaning: In the biological safety cabinet, take out the placenta from the placenta sample collection box, put it in a stainless steel cup, and use alcohol (75% alcohol) and normal saline to rinse the surface repeatedly (alcohol soaking disinfection time 30s)) , disinfect the placenta; then use surgical forceps to slowly tear off the fetal membranes, and place them in a glass dish. After removing the blood vessels, use phosphate buffer (0.025M sodium dihydrogen phosphate buffer at pH 6.5) to rinse again , remove surface dirt and impurities;
[0052] (2) Digestion treatment: in another cell culture plate, the fetal membrane tissue obtained in step (1) is cut into tissue blocks (cubic blocks of about 0.1 cubic centimeters), and the tissue blocks are put into digestive enzyme solution (preparation method : Add 0.1g of type I collagenase to 100ml of DMEM-F12, then f...
Embodiment 3
[0057] Embodiment 3, biological characteristic identification of fetal membrane MSC
[0058] The following tests were carried out on the canine fetal membrane mesenchymal stem cells obtained in Example 1 and Example 2, and there was no difference in the test results of the two kinds of stem cells.
[0059] 1. Cell growth and morphological characteristics
[0060] Through the isolation and culture of Example 1 and Example 2, the obtained canine fetal mesenchymal stem cells can be clearly seen under the microscope after 48 hours of culture. Spindle-shaped adherent cells will form turbine-shaped cell clones in about 8 days. After digestion and passage An adherent layer with about 75% fusion will be formed. During the culture process, it was found that the cell shape was relatively uniform, the proliferation speed was fast, the adhesion speed was fast, and it was easily digested by trypsin. After passage to 3-15 generations, its shape and growth characteristics did not change s...
PUM
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Description
Claims
Application Information
- IPC
- C12N5/0775; A01N1/02; A61K35/28; A61P19/02; A61P19/08; A61P21/00; A61P19/04; A61P25/28; A61P37/02; A61P27/02; A61P1/16; A61P9/00; A61P13/12; A61P3/10; A61P1/00; A61P5/14; A61P17/00
- CPC
- A01N1/0221; A61K35/28; A61P1/00; A61P1/16; A61P3/10; A61P5/14; A61P9/00; A61P13/12
- Inventors
- 王小武; 迟大明