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Panda placenta mesenchymal stem cell separation and culture method

A technology of mesenchymal stem cells and culture methods, applied in the field of separation and culture of giant panda placental mesenchymal stem cells, can solve the problems of difficult migration, difficulty in obtaining high-purity giant panda placental mesenchymal stem cells, polluting cells, etc., to reduce pollution The effect of the phenomenon

Pending Publication Date: 2022-07-08
秦岭大熊猫研究中心(陕西省珍稀野生动物救护基地)
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Problems solved by technology

[0003] However, after the giant panda placenta is collected, it is often difficult to obtain high-purity giant panda placental mesenchymal stem cells due to pollution and the difficulty of cell migration during tissue block culture.

Method used

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  • Panda placenta mesenchymal stem cell separation and culture method
  • Panda placenta mesenchymal stem cell separation and culture method
  • Panda placenta mesenchymal stem cell separation and culture method

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Embodiment

[0040] Refer to the attached figure 1 , is a flow chart of the method of the present invention, a method for separating and culturing giant panda placental mesenchymal stem cells, comprising the following steps:

[0041] Step 1: Aseptic collection of giant panda placenta, the specific operations are as follows:

[0042] Step 1.1: The giant panda placenta after natural childbirth was stored in normal saline containing 1% double antibody at 25°C, and transported to the laboratory within 6 hours for primary cell isolation;

[0043] Step 1.2: Transfer the placental tissue to a petri dish with 75% alcohol and soak it for 30s on an ultra-clean workbench. After taking it out, rinse it with normal saline for 5 to 10 times, and then immerse it in a normal saline containing 3% to 5% of tertiary antibodies for washing. For 1 min, transfer the placental tissue to a new petri dish, and repeat immersion and washing with the above-mentioned physiological saline containing 3% to 5% of the te...

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Abstract

The invention belongs to the field of stem cell separation and culture, and particularly relates to a panda placenta mesenchymal stem cell separation and culture method. Comprising the following steps: step 1, aseptically collecting panda placentas; 2, carrying out primary isolation on the giant panda placenta mesenchymal stem cells by adopting a collagenase digestion method, and culturing the separated tissues in a culture dish; 3, performing cell morphological characteristic identification, surface marker identification and induced differentiation identification on the placenta mesenchymal stem cells cultured in the step 2; and 4, cryopreserving the cultured and subcultured placenta mesenchymal stem cells, and taking out the placenta mesenchymal stem cells for cell resuscitation when the placenta mesenchymal stem cells are used. According to the method, high-purity giant panda placenta mesenchymal stem cells are obtained by adding high-concentration combined antibiotics and cleaning for multiple times when placentas are collected, performing isolated culture by adopting a collagenase digestion method, designing marker gene specific primers of giant panda mesenchymal stem cells and the like.

Description

technical field [0001] The invention belongs to the field of stem cell separation and culture, in particular to a method for separation and culture of giant panda placental mesenchymal stem cells. Background technique [0002] The giant panda is the flagship species of biodiversity conservation in the world. The Qinling giant panda is an important subspecies population of giant panda with unique morphological and genetic characteristics. Actively carry out the preservation of the Qinling giant panda germplasm resources and establish the Qinling giant panda germplasm. The resource stem cell bank plays an important role and significance in protecting the genetic diversity of giant pandas in my country. As pluripotent stem cells, mesenchymal stem cells have the potential of multi-directional differentiation, and can be used as ideal stem cells for clinical treatment, providing the best raw materials for the clinical treatment of giant panda diseases. The placenta, as a waste pr...

Claims

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Application Information

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IPC IPC(8): C12N5/0775C12Q1/02C12Q1/6888C12Q1/6851A01N1/02
CPCC12N5/0668G01N33/5005C12Q1/6888C12Q1/6851A01N1/0284C12N2509/00
Inventor 林鹏飞雷颖虎赵鹏鹏张丹辉沈洁娜赵卫平高更更
Owner 秦岭大熊猫研究中心(陕西省珍稀野生动物救护基地)
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