Separation method of human placenta mesenchymal stem cells

A technology of mesenchymal stem cells and separation methods, applied in the direction of animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve problems such as difficulty in obtaining a large number of pure placental mesenchymal stem cells and complex components of placental cells

Inactive Publication Date: 2015-10-28
大连金玛健康产业发展有限公司
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Problems solved by technology

[0003] In the prior art, the cell components of the placenta are relatively complex, especially the placental mesenchymal stem cells therein have multi-lineage differentiation potential and the cha

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  • Separation method of human placenta mesenchymal stem cells
  • Separation method of human placenta mesenchymal stem cells
  • Separation method of human placenta mesenchymal stem cells

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Embodiment Construction

[0017] The separation method of human placental mesenchymal stem cells is characterized in that comprising the following steps:

[0018] (1) Placental specimen processing, take the placental tissue produced at term under sterile conditions, cut small pieces of fetal decidua side placental tissue, cut into pieces, soak in the preservation solution containing antimicrobial peptides, and the preservation solution containing antimicrobial peptides It is prepared by adding 50 micrograms / ml polylysine to 50 micrograms / ml gallate EGCG. After soaking for a predetermined time, wash the blood stains repeatedly with normal saline until the washing solution is nearly colorless, and the treated placenta is obtained. tissue; the placental tissue was digested, with 15mmol / L Tris, adding Hartmann-D solution, adjusting the pH to 8.0, as a digestion buffer, adding a final concentration of 2.4g / L HyQTase and 300U / 200mL of the digestion solution composed of DNase I was removed, and the washed pl...

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Abstract

The invention discloses a separation method of human placenta mesenchymal stem cells. The separation method comprises the following steps that placenta specimen treatment is carried out, treated placenta tissue is obtained, tissue digestion is carried out on the placenta tissue, and cell suspension is obtained; preliminary subculture of the placenta mesenchymal stem cells is carried out; further separation and culture of the placenta mesenchymal stem cells are carried out. HyQTase and DNAse I are adopted for digestion together, the placenta mesenchymal stem cells are obtained through separation, after preliminary culture, microgravity treatment and electromagnetic field and sound wave treatment are carried out, BMP4 is used for treatment many times, upper layers are removed through a differential attachment method, a serum-free medium and antioxidant are replenished in a culture bottle for culture, and the final purity is high. The stem cell characteristics of the cells are verified through detection on hereditary stability, cell surface molecule expression conditions and cellular morphology in the continuous passage process, and materials are provided for seed cells with the placenta mesenchymal stem cells as clinical application.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for separating human placental mesenchymal stem cells. Background technique [0002] Placental mesenchymal stem cells have the characteristics of convenient collection, easy in vitro culture, expansion and induction, and are considered to be an ideal seed cell for stem cell research. It is usually difficult to obtain a large number of placental mesenchymal stem cells with high purity by conventional separation methods; although the cells obtained by flow cytometry or magnetic bead sorting are of good purity, the cost is high. The abundance of placental mesenchymal stem cells in placental tissue is low, and a lot of placental tissue is needed to obtain a certain number of primary cells. [0003] In the prior art, the cell components of the placenta are relatively complex, especially the placental mesenchymal stem cells therein have multi-lineage differentiation poten...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 夏凡
Owner 大连金玛健康产业发展有限公司
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