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Stem cell supernate extraction method

An extraction method and supernatant technology, applied in the field of stem cell supernatant extraction, can solve the problems of inconvenient acquisition of adipose-derived mesenchymal stem cells, secretion of various cytokines, and reduced extraction efficiency, so as to maintain the stability of passage and shorten the Extraction time, effect of insoluble tissue mass reduction

Pending Publication Date: 2022-04-01
上海泽充生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, there are still some problems in the existing stem cell supernatant extraction method: it is inconvenient to obtain adipose-derived mesenchymal stem cells, the extraction efficiency is reduced, and it is not conducive to the secretion of various cytokines by stem cells. Therefore, we propose a stem cell supernatant extraction method. method

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  • Stem cell supernate extraction method

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Embodiment 1

[0032] see figure 1 , the present invention provides a technical solution: a stem cell supernatant extraction method, comprising the following steps:

[0033] S1. Obtaining mesenchymal stem cells: obtaining mesenchymal stem cells from adipose tissue;

[0034] S2. Serum-free culture: use serum-free medium to culture mesenchymal stem cells;

[0035] S3. Extract the initial supernatant: when the cells are covered with a flat bottom, collect the culture medium, centrifuge, remove cell debris, and obtain the initial supernatant;

[0036] S4. Centrifugation: take the initial supernatant and further gradient centrifuge at 1800g for 8 minutes to remove dead cells and large debris to obtain the first supernatant after centrifugation;

[0037] S5. Cell extraction: after digesting cell fragments, dead cells and large fragments with digestive fluid, ultrasonic crushing, and then further gradient centrifugation at 1800g for 8 minutes to obtain the second supernatant;

[0038] S6. Mixing...

Embodiment 2

[0052] see figure 1 , the present invention provides a technical solution: a stem cell supernatant extraction method, comprising the following steps:

[0053] S1. Obtaining mesenchymal stem cells: obtaining mesenchymal stem cells from adipose tissue;

[0054] S2. Serum-free culture: use serum-free medium to culture mesenchymal stem cells;

[0055] S3. Extract the initial supernatant: when the cells are covered with a flat bottom, collect the culture medium, centrifuge, remove cell debris, and obtain the initial supernatant;

[0056] S4. Centrifugation: take the initial supernatant and further gradient centrifuge at 2400g for 12 minutes to remove dead cells and large debris to obtain the first supernatant after centrifugation;

[0057] S5. Cell extraction: after digesting cell fragments, dead cells and large fragments with digestive fluid, ultrasonic crushing, and then further gradient centrifugation at 2400g for 12 minutes to obtain the second supernatant;

[0058] S6. Mixi...

Embodiment 3

[0072] see figure 1 , the present invention provides a technical solution: a stem cell supernatant extraction method, comprising the following steps:

[0073] S1. Obtaining mesenchymal stem cells: obtaining mesenchymal stem cells from adipose tissue;

[0074] S2. Serum-free culture: use serum-free medium to culture mesenchymal stem cells;

[0075] S3. Extract the initial supernatant: when the cells are covered with a flat bottom, collect the culture medium, centrifuge, remove cell debris, and obtain the initial supernatant;

[0076] S4. Centrifugation: take the initial supernatant and further gradient centrifuge at 2200g for 10 min to remove dead cells and large debris to obtain the first supernatant after centrifugation;

[0077] S5. Cell extraction: after digesting cell fragments, dead cells and large fragments with digestive fluid, ultrasonic crushing, and then further gradient centrifugation at 2200g for 10 minutes to obtain the second supernatant;

[0078] S6. Mixing, ...

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Abstract

The invention discloses a stem cell supernate extraction method. The stem cell supernate extraction method comprises the following steps: S1, acquiring mesenchymal stem cells: acquiring the mesenchymal stem cells from adipose tissues; s2, serum-free culture: culturing the mesenchymal stem cells by adopting a serum-free culture medium; s3, extracting an initial supernatant: collecting the culture medium after the cells grow to be full of flat bottoms, centrifuging, and removing cell debris to obtain the initial supernatant; s4, performing centrifugation: taking the initial supernatant, performing further gradient centrifugation on the initial supernatant for 1800-2400g for 8-12min, and removing dead cells and large fragments to obtain a first supernatant after centrifugation; s5, cell extraction: digesting the cell debris, dead cells and large debris with digestive juice, then carrying out ultrasonication, and then carrying out gradient centrifugation for 8-12min at 1800-2400g to obtain a second supernatant; and S6, mixing, filtering and concentrating. In the process of obtaining the mesenchymal stem cells, the extraction time of the adipose-derived mesenchymal stem cells can be shortened, the extraction efficiency of the adipose-derived mesenchymal stem cells is effectively improved, and meanwhile, the passage stability is kept.

Description

technical field [0001] The invention belongs to the technical field of supernatant liquid extraction, and in particular relates to a stem cell supernatant liquid extraction method. Background technique [0002] Adipose stem cells are a kind of stem cells with multi-directional differentiation potential isolated from adipose tissue in recent years. Under certain induction environment, they can differentiate into bone cells, chondrocytes, cardiomyocytes, neuron cells, etc. Adipose stem cells have the advantages of convenient extraction and wide distribution. They can be used as seed cells for cell transplantation in the fields of tissue engineering and regenerative medicine, and have very important research and application values. [0003] At present, there are still some problems in the existing stem cell supernatant extraction method: it is inconvenient to obtain adipose-derived mesenchymal stem cells, the extraction efficiency is reduced, and it is not conducive to the secr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 李一佳
Owner 上海泽充生物技术有限公司
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