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Large-scale preparation method of placenta mesenchymal stem cells

A technology of mesenchymal stem cells and placenta, applied in the field of preparation of placental mesenchymal stem cells, can solve the problems of large volume of placental tissue, easy to wash incompletely, and many reagents and equipment

Inactive Publication Date: 2010-12-22
青岛奥克生物开发有限公司
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Problems solved by technology

[0005] Extract mesenchymal stem cells from placenta tissue. The blood content in the placenta is very large, and it consumes a lot of reagents when washing, and it is easy to wash incompletely; now the whole placenta tissue is cut, and the placenta tissue is large and thick, and it takes time to cut. Time-consuming and labor-intensive; the use of digestive enzymes for digestion requires a large amount of enzymes and a lot of consumables; due to the large volume and quantity of tissue blocks during tissue inoculation, a lot of reagents and equipment are consumed, and the workload is heavy and easy to contaminate

Method used

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  • Large-scale preparation method of placenta mesenchymal stem cells
  • Large-scale preparation method of placenta mesenchymal stem cells
  • Large-scale preparation method of placenta mesenchymal stem cells

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Embodiment Construction

[0026] Preparation of placental amnion-derived mesenchymal stem cells:

[0027] 1. Wash the placenta with the PBS buffer solution containing 100kU / L penicillin+100mg / L streptomycin+2U / ml heparin sodium (Heparin Sodium, Qingdao Kangyuan Pharmaceutical (Group) Co., Ltd.), and place the amniotic membrane on the fetal surface along the edge of the placenta. Peel off slowly. The penicillin / streptomycin solution was purchased from ScienCell Inc., P / S (Penicillin / Streptomycin Solution).

[0028] 2. Wash the placenta-derived amniotic membrane again, and cut the amniotic membrane into 1-5mm 3 small pieces.

[0029] 3. Mix the amnion tissue block with DMEM medium containing 100kU / L penicillin and 100mg / L streptomycin, and centrifuge at 850g for 10min.

[0030] 4. Discard the supernatant, add digestion solution (DMEM medium containing 0.25% trypsin by volume) to each tube for digestion at 37° C. for 10 min, and centrifuge at 850 g for 10 min.

[0031] 5. Discard the supernatant, add ...

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Abstract

The invention relates to a preparation method of placenta mesenchymal stem cells. The method routinizes the preparation process, simplifies operation steps, shortens the operation time and reduces the workload, thereby preventing pollution to a great degree, reducing reagent and material required in the preparation process and increasing the yield of mesenchymal stem cells.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a preparation method of placental mesenchymal stem cells. technical background [0002] Mesenchymal stem cells have broad application prospects in developmental biology research and clinical treatment, such as tissue and organ repair. Mesenchymal stem cells were first confirmed in bone marrow, and then in adult peripheral blood, periosteum, muscle and adipose tissue. The presence of mesenchymal stem cells was found in other places. Mesenchymal stem cells are pluripotent stem cells, which can differentiate into different types of cells such as bone, fat, cartilage, muscle and liver cells under the corresponding induction conditions. At present, adult bone marrow is the main source of mesenchymal stem cells for clinical application, but there are obvious defects, such as the content of mesenchymal stem cells in adult bone marrow is very small (about 0.001%-0.01%). ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0735
Inventor 魏斯溧高宏王丽胡建霞张学峰张美荣
Owner 青岛奥克生物开发有限公司
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