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A method for obtaining pancreatic precursor cells and islet β cells by differentiating human pluripotent stem cells

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

Active Publication Date: 2021-04-02
深圳市北科源细胞科技有限公司
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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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  • A method for obtaining pancreatic precursor cells and islet β cells by differentiating human pluripotent stem cells
  • A method for obtaining pancreatic precursor cells and islet β cells by differentiating human pluripotent stem cells
  • A method for obtaining pancreatic precursor cells and islet β cells by differentiating human pluripotent 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, 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;

[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 the following working concen...

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Abstract

The invention provides a method for obtaining pancreatic precursor cells and islet beta cells by differentiating human pluripotent stem cells. During the specialization process of endoderm cells derived from human pluripotent stem cells to pancreatic lineage cells, the differentiation efficiency of human pluripotent stem cells to pancreatic precursor cells can be improved by adding WNT signaling pathway inhibitors in stages. After improving the differentiation efficiency of pancreatic precursor cells, the invention further improves the differentiation efficiency of mature pancreatic beta cells. This method is suitable for pancreatic differentiation of different cell lines to obtain a large number of islet β cells, which provides strong support for diabetes cell therapy, drug screening, disease models, etc., and 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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12N5/0735C12N5/10
CPCC12N5/0613C12N2506/45C12N2506/02C12N2501/415C12N2501/16C12N5/0676C12N5/0678C12N2500/02C12N2501/117
Inventor 蒋卫檀梦天
Owner 深圳市北科源细胞科技有限公司
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