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Preparation method of decidual mesenchymal stem cells

A technology of stem cells and decidua, applied in the field of stem cells, can solve problems such as being susceptible to pollution, and achieve the effect of high viability

Inactive Publication Date: 2016-12-07
HANGZHOU S EVANS BIOSCI LTD
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  • Claims
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Problems solved by technology

The published endometrial mesenchymal stem cells are basically isolated from women's menstrual blood, which has the advantages of convenient collection and unlimited number of times, but also has the disadvantage of being easily contaminated due to in vitro personal operations in a non-sterile environment. Therefore, , it is very meaningful to be able to isolate the source and method of endometrial mesenchymal stem cells with high quality and ensure sterility and safety

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  • Preparation method of decidual mesenchymal stem cells
  • Preparation method of decidual mesenchymal stem cells
  • Preparation method of decidual mesenchymal stem cells

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preparation example Construction

[0031] The preparation method of decidual mesenchymal stem cells provided by the invention comprises the following steps:

[0032] 1) Sampling and pretreatment: take decidua tissue, wash with saline, add 0.8-1.1 times the volume of collection solution, and store at low temperature;

[0033] 2) Cell separation: filter the sample treated in step 1) with sterile gauze, and perform virus detection on the filtrate. If the filtrate test result is negative, perform the following operations on the corresponding filter residue: wash twice with PBS, cut into pieces , transferred to a centrifuge tube, added an equal volume of 0.25% type Ⅰ collagenase for digestion for 30 minutes, then added an equal volume of Chang's complete medium to stop digestion, centrifuged, discarded the supernatant, washed with PBS, added Chang's complete medium to resuspend the cells, and obtained single cell suspension;

[0034] 2) Cell separation: filter the sample treated in step 1) with sterile gauze, and p...

Embodiment 1

[0041] A method for preparing decidual mesenchymal stem cells, comprising the following steps:

[0042] 1) Sampling and pre-treatment: take the decidual tissue, wash it with normal saline for 3 times, add an equal volume of collection solution, and store at 4-10°C;

[0043] 2) Cell separation: filter the sample treated in step 1) with sterile gauze, and carry out virus detection on the filtrate. If the filtrate test result is negative, perform the following operations on the corresponding filtered tissue: wash with PBS for 2 Once, use scissors to cut the tissue in the filter residue to 2mm 3 About the size, transfer it into a centrifuge tube, add an equal volume of 0.25% type Ⅰ collagenase to digest at 37°C for 30 minutes, then add an equal volume of Chang’s complete medium to stop the digestion, centrifuge at 1200rpm / min for 10min, discard the supernatant, and wash once with PBS , adding Chang's complete medium to resuspend the cells to obtain a single cell suspension;

[0...

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Abstract

The invention discloses a preparation method of decidual mesenchymal stem cells. The preparation method comprises the following steps of 1 sampling and pretreating, 2 cell separating, 3 cell passage culturing and 4 cryopreserving. According to the method, the decidual mesenchymal stem cells are prepared from decidual tissue, and the high-purity, high-viability and contamination-free stem cells are obtained on the basis that a mature, safe and reliable separation technology is established and preserved in liquid nitrogen for a long term, can be recovered to be used for stem cell transplanting and are expected to gradually replace bone marrow stem cells which are expensive and difficult to match.

Description

technical field [0001] The invention relates to the technical field of stem cells, in particular to a method for preparing decidual mesenchymal stem cells. Background technique [0002] Stem cell technology and its application are considered to be human engineering health engineering in the 21st century, and its research content involves almost all biomedical fields of life sciences. It has an important impact on new genes, gene function analysis, developmental biology models and new drug development. The clinical application of stem cell technology to refractory diseases will lead to revolutionary progress in the medical field. Stem cells currently being developed around the world are mainly mesenchymal stem cells. [0003] Mesenchymal stem cells are pluripotent stem cells derived from mature tissues, with high self-renewal ability and multi-directional differentiation potential, and can differentiate into osteoblasts, chondrocytes, liver cells, pancreatic islet cells, et...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0682C12N5/0668C12N2509/00
Inventor 全丽丽
Owner HANGZHOU S EVANS BIOSCI LTD
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