Method for transforming umbilical cord mesenchymal stem cells into islet cells

A technique for stem cells and islet cells, which is applied in the field of using umbilical cord mesenchymal stem cells to induce transformation into islet cells, can solve the problems of cumbersome process, low quantity, high cost, etc. Effect

Inactive Publication Date: 2015-10-21
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this co-culture method not only requires a certain source of islet cells, but the process is cumbersome and expensive, and more scattered islet-like cells are obtained, the number is small, there are almost no islet cell clusters, and scattered islet-like cells The cells are relatively small, and the amount of insulin secretion is small, so it is difficult to meet the requirements of clinical transplantation

Method used

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  • Method for transforming umbilical cord mesenchymal stem cells into islet cells
  • Method for transforming umbilical cord mesenchymal stem cells into islet cells
  • Method for transforming umbilical cord mesenchymal stem cells into islet cells

Examples

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Embodiment 1

[0037] The method for converting umbilical cord mesenchymal stem cells into islet cells provided by the present invention comprises the following steps:

[0038] (1) Isolation and culture of umbilical cord mesenchymal stem cells

[0039] Under sterile conditions, take the umbilical cord of a healthy fetus with full-term pregnancy and cesarean section. After the umbilical cord is isolated from the body, soak it in normal saline containing 25U heparin sodium per milliliter for 4-6 hours. Remove the outer amniotic membrane, cut the umbilical cord into 1mm×1mm×1mm tissue pieces and paste them on the bottom of the culture dish, add low-sugar DMEM containing 10% fetal bovine serum and place in a 37°C, 5% CO2 saturated humidity incubator Cultured in umbilical cord, removed after 2 weeks of culture, and subcultured when the adherent cells reached 80%-90% confluence, to obtain the third generation umbilical cord mesenchymal stem cells.

[0040] Among them, the composition of low-sugar...

Embodiment 2

[0048] Compared with Example 1, this example is different except for the step 1) of induction and transformation of the third-generation umbilical cord mesenchymal stem cells, and the other process steps are the same.

[0049] In this embodiment, the induction transformation step 1) of the third generation umbilical cord mesenchymal stem cells is:

[0050] Add the first induction solution containing 60 μg / L β-nerve growth factor, 1 nmol / L activin A, 15 mmol / L nicotinamide, 15 μg / L epidermal growth factor, and 15 μg / L β-mercaptoethanol, with a volume fraction of 5 % fetal bovine serum DMEM / F12 culture medium for 7 days until the cells complete the proliferation process to obtain pre-transformed stem cell bodies.

Embodiment 3

[0052] Compared with Example 1, this example is the same except for the step 1) of inducing transformation of umbilical cord mesenchymal stem cells.

[0053] In this embodiment, step 1) of induction and transformation of umbilical cord mesenchymal stem cells is:

[0054] Add the first induction solution containing 100 μg / L β-nerve growth factor, 4 nmol / L activin A, 10 mmol / L nicotinamide, 25 μg / L epidermal growth factor, and 5 μg / L β-mercaptoethanol at a volume fraction of 10%. The DMEM / F12 medium with fetal bovine serum was cultured for 7 days until the cells completed the proliferation process to obtain pre-transformed stem cell bodies.

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Abstract

The invention relates to a method for transforming umbilical cord mesenchymal stem cells into islet cells. The method comprises the steps that after the umbilical cord mesenchymal stem cells are separated under the aseptic condition and subcultured, the induction is conducted, wherein 1 beta-nerve growth factors, activin A, niacinamide and epidermal growth factors are added into a DMEM / F12 culture medium containing fetal calf serum for conducting the induction culture; 2 a DMEM / F12 culture medium containing niacinamide, alkaline fibroblast growth factors and insulin-transferrin-selenium is added in for continuous culture till the cell transformation is ended, and islet cell sap is obtained. By means of the method, the number of the obtained islet cells is large, the quality of the islet cells is good, more insulin can be secreted, and the method is of great importance in clinic transformation of the umbilical cord mesenchymal stem cells for treating the 1 type diabetes.

Description

technical field [0001] The invention relates to the fields of bioengineering, tissue engineering and biomedicine, in particular to a method for inducing transformation of umbilical cord mesenchymal stem cells into islet cells. Background technique [0002] Diabetes mellitus is a disorder of sugar, fat and protein metabolism caused by relative or absolute deficiency of insulin secretion or insulin receptor defects. Clinically, type Ⅰ and some type Ⅱ diabetes are mostly treated with subcutaneous injection of insulin. Cell therapy is an ideal treatment strategy for diabetes, especially type 1 diabetes. Especially for diabetic patients who have lost the function of islet cells, implanting insulin-secreting islet cells or their substitutes is the most ideal treatment method. At present, pancreatic islet cell transplantation has achieved some curative effects in the treatment of diabetes, but difficulties such as insufficient source of donor cells and severe immune rejection grea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 曾宪卓鲁菲
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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