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Mesenchymal stem cell differentiation induction liquid and method for inducing mesenchymal stem cell to adipose cells

A bone marrow mesenchymal and stem cell differentiation technology, applied in the field of cell engineering, can solve the problem that there is no differentiation induction solution for finished products, and achieve the effect of high fusion

Inactive Publication Date: 2017-03-08
深圳市艾克希尔生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a bone marrow mesenchymal stem cell differentiation induction solution and a method for inducing it into adipocytes, solving the problem that there is no finished differentiation induction solution in the prior art, which can differentiate bone marrow mesenchymal stem cells Problems induced into fat cells

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  • Mesenchymal stem cell differentiation induction liquid and method for inducing mesenchymal stem cell to adipose cells
  • Mesenchymal stem cell differentiation induction liquid and method for inducing mesenchymal stem cell to adipose cells
  • Mesenchymal stem cell differentiation induction liquid and method for inducing mesenchymal stem cell to adipose cells

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Embodiment 1

[0041] 1) The bone marrow was extracted and cultured by the direct adherence method, and the unattached cells were discarded during the culture process.

[0042] 2) Digest the adherent cells with trypsin for 5 minutes, subculture the digested bone marrow mesenchymal stem cells; use DMEM / F12 mesenchymal stem cell culture medium for direct adherent culture, at 37°C, 5% CO 2 Culture in the incubator, change the medium after 24 hours, discard the unattached cells, and change the medium every 3 days after that, pay attention to observe the shape and growth of the cells during the period. When trypsinizing adherent cells, stop the digestion with medium when the cells start to round.

[0043] 3) Obtain bone marrow mesenchymal stem cells with high fusion degree after subculture for the third generation.

[0044] 4) Add a differentiation-inducing solution to the bone marrow mesenchymal stem cells with a high degree of confluence for induction and culture for 48 hours; the differentia...

Embodiment 2

[0050] 1) The bone marrow was extracted and cultured by the direct adherence method, and the unattached cells were discarded during the culture process.

[0051] 2) Digest the adherent cells with trypsin for 10 minutes, subculture the digested bone marrow mesenchymal stem cells; use DMEM / F12 mesenchymal stem cell culture medium for direct adherent culture, at 37°C, 5% CO 2 Culture in an incubator, change the medium after 24 hours, discard unattached cells, and change the medium every 2 days thereafter, pay attention to observe the shape and growth of the cells during the period. When trypsinizing adherent cells, stop the digestion with medium when the cells start to round.

[0052] 3) Obtain bone marrow mesenchymal stem cells with high fusion degree after subculture for the third generation.

[0053] 4) Add differentiation-inducing liquid to the highly confluent bone marrow mesenchymal stem cells for induction and culture for 96 hours; the differentiation-inducing liquid inc...

Embodiment 3

[0059] 1) The bone marrow was extracted and cultured by the direct adherence method, and the unattached cells were discarded during the culture process.

[0060] 2) Digest the adherent cells with trypsin for 7 minutes, subculture the digested bone marrow mesenchymal stem cells; use DMEM / F12 mesenchymal stem cell culture medium for direct adherent culture, at 37°C, 5% CO 2 Culture in the incubator, change the medium after 24 hours, discard the unattached cells, and change the medium every 3 days after that, pay attention to observe the shape and growth of the cells during the period. When trypsinizing adherent cells, stop the digestion with medium when the cells start to round.

[0061] 3) Obtain bone marrow mesenchymal stem cells with high fusion degree after subculture for the third generation.

[0062] 4) Add differentiation-inducing liquid to the highly confluent bone marrow mesenchymal stem cells for induction and culture for 72 hours; the differentiation-inducing liquid...

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Abstract

The invention relates to mesenchymal stem cell differentiation induction liquid and a method for inducing mesenchymal stem cells to adipose cells. The mesenchymal stem cell differentiation induction liquid is prepared from the following components in parts by mass: 0.05mmol / L to 0.2mmol / L of indomethacin, 0.1mmol / L to 0.5mmol / L of 3-isobutyl-1-methylxanthine, 10<-4>mol / L to 10<-6>mol / L of dexamethasone and 5mg / L to 20mg / L of insulin. The mesenchymal stem cell differentiation induction liquid provided by the invention is used for inducing and differentiating the mesenchymal stem cells; after 48 hours, less cytoplasm contains lipid granules with relatively high refractivity can be seen; when culture time is prolonged, the small lipid granules are gathered and gradually become large to form lipid droplets; the quantity of the adipose cells is gradually increased, and round and red lipid droplets with different sizes exist in the mesenchymal stem cells treated by an oil red O dyeing agent.

Description

technical field [0001] The invention belongs to the technical field of cell engineering, and more specifically relates to a bone marrow mesenchymal stem cell differentiation induction solution and a method for inducing bone marrow mesenchymal stem cells into adipocytes. Background technique [0002] Adult stem cells in the mesoderm are the main source of mesenchymal stem cells (MSCs). Bone marrow mesenchymal stem cells (hMSCs), as an important component of the hematopoietic microenvironment, are induced under different conditions in vitro. It can differentiate into tissue cells in multiple directions such as osteoblasts, chondrocytes, fat cells, nerve cells, liver cells, fibroblasts, and tendon cells. Studies have found that bone mass and fat content in the bone marrow of patients with osteoporosis are inversely proportional to each other. Bone mass loss increases fat content. It is speculated that osteoporosis may be caused by the imbalance of bone marrow stem cells and the...

Claims

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Application Information

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IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2500/30C12N2500/40C12N2501/30C12N2501/33C12N2506/1353
Inventor 李珍珠
Owner 深圳市艾克希尔生物科技有限公司
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