Induction culture medium and method for inducing human fat mesenchymal stem cells to generate insulin secreting cells

A technique for inducing medium and mesenchymal stem cells, which is applied in the field of inducing medium and can solve problems such as unsatisfactory inducing effect

Active Publication Date: 2015-01-07
胡振波
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Problems solved by technology

[0007] The technical problem to be solved in the present invention is to provide an induction medium for the above deficiencies, which overcomes the de

Method used

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  • Induction culture medium and method for inducing human fat mesenchymal stem cells to generate insulin secreting cells
  • Induction culture medium and method for inducing human fat mesenchymal stem cells to generate insulin secreting cells

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Embodiment 1

[0059] Embodiment 1, a method for inducing human adipose-derived mesenchymal stem cells to generate insulin-secreting cells, comprising the following steps:

[0060] Isolation and culture of human adipose-derived mesenchymal stem cells

[0061] (1) Collect 100ml of adipose tissue from diabetic patients, use ophthalmic scissors to cut as much as possible the small blood vessels visible to the naked eye in the adipose tissue, wash the blood, cut up the adipose tissue, transfer it to a centrifuge tube, add 0.25% trypsin and 0.1% collagenase The mixed solution, the volume ratio of 0.25% trypsin and 0.1% collagenase in the mixed solution is 1:1, and the consumption of the mixed solution is twice the volume of the fat tissue;

[0062] (2) Use a vortex shaker to vibrate the above adipose tissue suspension at 1500 rpm for 60 seconds;

[0063] (3) at 37 o Digest in a constant temperature shaker with shaking for 1 hour. The liquid surface is divided into three layers. The upper layer ...

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Abstract

The invention discloses an induction culture medium which comprises a solvent DMEM (dulbecco's modified eagle medium)-high sugar culture medium, platelets, all-trans vitamin A acid, activin A, glicetin I, a fibroblast growth factor, beta cytosine, an insuline-like growth factor, nicotinamide, polypeptide, pentagastrin and mercaptoethanol. Physical oscillation and collagenase/pancreatin dual digestion are performed to efficiently separate fat mesenchymal stem cells from human fat tissues. The fat mesenchymal stem cells are cultured for proliferation and subcultivation, and the induction culture medium is utilized to induce the fat mesenchymal stem cells to become insulin secreting cells, so that more than 40% of the fat stem cells are induced to differentiate into the insulin secreting cells.

Description

technical field [0001] The invention relates to an induction medium, in particular to an induction medium and a method for inducing human adipose-derived mesenchymal stem cells to generate insulin-secreting cells, and belongs to the field of medical technology. Background technique [0002] The prevalence of diabetes in my country has exceeded 12%. About 200 million people worldwide suffer from diabetes in varying degrees. The pathogenesis of diabetes is mainly due to the damage and dysfunction of pancreatic islet secretory cells, or the abnormal immune function of the body, which leads to absolute or relative deficiency of insulin secretion, which leads to elevated blood sugar, which in turn causes damage to various organs of the body and various complications. Clinical treatment usually uses subcutaneous injection of insulin to lower blood sugar, but for patients with type I and some type II diabetes, the effect of this treatment method is not ideal, because it is easy to...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/0775
Inventor 胡振波
Owner 胡振波
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