Culture medium for inducing human umbilical cord mesenchymal stem cells to differentiate into islet-like cells and induction method therefor

A technology of islet-like cells and stromal cells, applied in the field of islet-like cells, can solve the problems of many interference factors, irregular induction factors, uncertain induction scheme, etc., and achieve broad clinical application prospects, increase induction efficiency, and high insulin expression. Effect

Pending Publication Date: 2016-06-15
王意忠
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, how to efficiently induce their differentiation into islet-like cells for cell biotherapy still has many problems to be solved: 1) The differentiation of stem cells into islet-like cells is subject to many interference factors, first of all, the induction scheme is uncertain; 2) ) The induction factors used are not standardized, which cannot guarantee the differentiation and proliferation of stem cells in the expected direction; 3) β-mercaptoethanol is often used, and its toxicity is prohibitive, which affects subsequent clinical applications; 4) Insulin induced differentiation of islet-like cells Low secretion efficiency

Method used

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  • Culture medium for inducing human umbilical cord mesenchymal stem cells to differentiate into islet-like cells and induction method therefor
  • Culture medium for inducing human umbilical cord mesenchymal stem cells to differentiate into islet-like cells and induction method therefor
  • Culture medium for inducing human umbilical cord mesenchymal stem cells to differentiate into islet-like cells and induction method therefor

Examples

Experimental program
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Effect test

Embodiment 1

[0060] Example 1: A culture medium for inducing human umbilical cord mesenchymal stem cells to differentiate into fat-like cells contains the following composition:

[0061] Activin A4nmol / L

[0062] Epidermal growth factor 25μg / L

[0063] β-nerve growth factor 100μg / L

[0064] Niacinamide 10mmol / L

[0065] Insulin-Transferrin-Selenium 1%

[0066] Basic fibroblast growth factor 10 μg / L

[0067] UItraCULTURE medium balance.

[0068] Preparation of the above-mentioned medium: firstly, activin A, epidermal growth factor, β-nerve growth factor, nicotinamide, and basic fibroblast growth factor were added to UItraCULTURE medium (commercially available UItraCULTURE medium), so that the final The concentrations are 4nmol / L, 25μg / L, 100μg / L, 10mmol / L and 10μg / L respectively. Add 1% insulin-transferrin-selenium to the above solution and incubate at 37°C for half an hour.

Embodiment 2

[0069] Embodiment 2: A culture medium for inducing human umbilical cord mesenchymal stem cells to differentiate into fat-like cells contains the following composition:

[0070] Activin A2nmol / L

[0071] Epidermal growth factor 20μg / L

[0072] β-nerve growth factor 80μg / L

[0073] Niacinamide 5mmol / L

[0074] Insulin-Transferrin-Selenium 0.5%

[0075] Basic fibroblast growth factor 5 μg / L

[0076] UItraCULTURE medium balance.

[0077] Preparation of the above-mentioned medium: firstly, activin A, epidermal growth factor, β-nerve growth factor, nicotinamide, and basic fibroblast growth factor were added to UItraCULTURE medium (commercially available UItraCULTURE medium), so that the final The concentrations were 2nmol / L, 20μg / L, 80μg / L, 5mmol / L and 5μg / L, and 0.5% insulin-transferrin-selenium was added to the above solution and incubated at 37°C for half an hour.

Embodiment 3

[0078] Example 3: A culture medium for inducing human umbilical cord mesenchymal stem cells to differentiate into fat-like cells contains the following composition:

[0079] Activin A6nmol / L

[0080] Epidermal growth factor 30μg / L

[0081] β-nerve growth factor 120μg / L

[0082] Niacinamide 15mmol / L

[0083] Insulin-Transferrin-Selenium 1.5%

[0084] Basic fibroblast growth factor 15μg / L

[0085] UItraCULTURE medium balance.

[0086] Preparation of the above-mentioned medium: firstly, activin A, epidermal growth factor, β-nerve growth factor, nicotinamide, and basic fibroblast growth factor were added to UItraCULTURE medium (commercially available UItraCULTURE medium), so that the final Concentrations are 6nmol / L, 30μg / L, 120μg / L, 15mmol / L and 15μg / L respectively. Add 1.5% insulin-transferrin-selenium to the above solution and incubate at 37°C for half an hour.

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Abstract

The invention discloses a medium for inducing human umbilical cord mesenchymal stem cells to differentiate into islet-like cells and an inducing method thereof. The medium contains activin A, epidermal growth factor, β-nerve growth factor, nicotinamide, 1% insulin-transferrin-selenium, and 10 μg/L basic fibroblast growth factor UltraCULTURE medium. The induction method is as follows: the mesenchymal stem cells are induced, differentiated and cultured using the above-mentioned induction medium. The induction medium of the invention contains simple induction factors, does not contain toxic and harmful substances, and is safer. The method of the present invention for inducing human umbilical cord mesenchymal stem cells to differentiate into islet-like cells can obtain aggregated islet-like cells in 21 days, and partially mature and differentiated islet-like cells can be obtained in 28 days. A standardized and novel induction method has been established. Broad clinical application prospects.

Description

technical field [0001] The invention belongs to the field of stem cells, in particular to a method for inducing human umbilical cord mesenchymal stem cells to differentiate into islet-like cells. [0002] Cell culture medium and its induction method. Background technique [0003] Diabetes mellitus is a metabolic syndrome characterized by hyperglycemia and glucosuria due to defects in insulin secretion or insulin action. Patients' persistent high blood sugar and long-term metabolic disorders can lead to damage and dysfunction of various systems in the body. The clinical treatment of diabetes includes conventional treatments such as diabetes education, exercise therapy, diet therapy, hypoglycemic drugs, and insulin therapy. However, the current treatment effect is not satisfactory. Therefore, it is necessary to find new islet substitutes for the treatment of diabetes. Allogeneic pancreas transplantation has a poor effect, complex operation, high mortality rate, and severe tr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 王意忠崔晓兰时瀚申义山霞
Owner 王意忠
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