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Method for obtaining pancreatic precursor cells and pancreatic beta cells through differentiation of human multipotential stem cells

A technology of human pluripotent stem cells and pancreatic precursor cells, which is applied in the field of stem cells and regenerative medicine, can solve the problems of inability to obtain stable and reproducible islet cell differentiation schemes, and achieve the effect of improving the transcription level

Active Publication Date: 2019-05-14
SHENZHEN BEIKE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Due to the complexity of the differentiation of pancreas and islet β cells and the differences between different systems, it is difficult to obtain a reproducible and efficient islet cell differentiation plan, and it is impossible to obtain a stable and large number of pancreatic precursor cells

Method used

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  • Method for obtaining pancreatic precursor cells and pancreatic beta cells through differentiation of human multipotential stem cells
  • Method for obtaining pancreatic precursor cells and pancreatic beta cells through differentiation of human multipotential stem cells
  • Method for obtaining pancreatic precursor cells and pancreatic beta cells through differentiation of human multipotential stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Differentiation of human embryonic stem cells into pancreatic precursor cells and mature islet β cells under the induction of WNT signaling pathway inhibitor XAV-939

[0057] (1) Cell differentiation

[0058] 1) Differentiation of human embryonic stem cells into definitive endoderm cells:

[0059] a. Preparing culture medium 1 for definitive endoderm stage, and culturing human embryonic stem cells for 1 day in a carbon dioxide incubator at 37 degrees Celsius using the culture;

[0060] b. Prepare definitive endoderm stage medium 2, replace the cells cultured in the above step a with definitive endoderm stage medium 2, and culture in a carbon dioxide incubator at 37 degrees Celsius for 3 days, and replace the medium every day;

[0061] The composition of the definitive endoderm stage culture medium 1 is: mix the IMDM medium and the F12 medium at a ratio of 1:1 to make the basal medium, and also include the following working concentration components: 0.2% bovin...

Embodiment 2

[0088] Example 2 Differentiation of human embryonic stem cells into pancreatic precursor cells and islet β cells under the induction of WNT signaling pathway inhibitor IWR-1

[0089] (1) Cell differentiation

[0090] 1) Differentiation of human embryonic stem cells into definitive endoderm cells:

[0091] a. Preparing culture medium 1 for definitive endoderm stage, and culturing human embryonic stem cells for 1 day in a carbon dioxide incubator at 37 degrees Celsius using the culture;

[0092] b. Prepare definitive endoderm stage medium 2, replace the cells cultured in the above step a with definitive endoderm stage medium 2, and culture in a carbon dioxide incubator at 37 degrees Celsius for 3 days, and replace the medium every day;

[0093] The composition of the definitive endoderm stage culture medium 1 is: mix the IMDM medium and the F12 medium at a ratio of 1:1 to make the basal medium, and also include the following working concentration components: 0.2% bovine serum a...

Embodiment 3

[0120] Example 3 Differentiation of human induced pluripotent stem cells into pancreatic precursor cells and mature islet β cells under the induction of WNT signaling pathway inhibitor XAV-939

[0121] (1) Cell differentiation

[0122] 1) Differentiation of human induced pluripotent stem cells into definitive endoderm cells:

[0123] a. Preparing culture medium 1 for definitive endoderm stage, and culturing human induced pluripotent stem cells for 1 day in a carbon dioxide incubator at 37 degrees Celsius using the culture;

[0124] b. Prepare definitive endoderm stage medium 2, which will go through the above steps a The cultured cells were replaced with definitive endoderm stage medium 2, cultured in a carbon dioxide incubator at 37 degrees Celsius for 3 days, and the medium was changed every day;

[0125] The composition of the definitive endoderm stage culture medium 1 is: mix the IMDM medium and the F12 medium at a ratio of 1:1 to make the basal medium, and also include ...

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Abstract

The invention provides a method for obtaining pancreatic precursor cells and pancreatic beta cells through differentiation of human multipotential stem cells. In the process of specializing entoderm cells derived from the human multipotential stem cells towards pancreas pedigree cells, the efficiency of differentiating the human multipotential stem cells towards the pancreatic precursor cells canbe improved by stepwise adding a WNT signal channel inhibitor. According to the method, after the efficiency of differentiation towards the pancreatic precursor cells is improved, the efficiency of differentiation of the mature pancreatic beta cells is improved accordingly. The method is suitable for pancreas differentiation of different cell lines, a large quantity of pancreatic beta cells are obtained, strong support is provided for diabetes cell therapy, drug screening, disease models and the like, and the method has a good application prospect.

Description

technical field [0001] The invention belongs to the field of stem cells and regenerative medicine, and relates to a method for obtaining pancreatic precursor cells and islet beta cells by differentiating human pluripotent stem cells. Background technique [0002] Diabetes is a metabolic disease characterized by elevated blood sugar. Whether it is the loss of β cells caused by immune injury leading to insufficient insulin secretion (T1D), or the increase in blood sugar caused by insulin resistance (T2D), eventually there will be damage to β cells (Lam and Cherney, 2018). According to the report of the World Diabetes Federation IDF in 2017, there are 425 million diabetic patients today, and this huge sick population may increase to 629 million in 2045. Diabetes is no longer a health risk, it has become a global social crisis. Diabetes not only causes blood sugar to rise, but also is accompanied by complications caused by diabetes, which brings various burdens to the family a...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/0735C12N5/10
CPCC12N5/0613C12N2506/45C12N2506/02C12N2501/415C12N2501/16C12N5/0676C12N5/0678C12N2500/02C12N2501/117
Inventor 蒋卫檀梦天
Owner SHENZHEN BEIKE BIOTECH
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