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Induction method for promoting conversion of pancreas islet alpha cells into beta cells and application of induction method

A technology of β cells and pancreatic islets, which is applied in the field of biomedicine to achieve the effect of increasing the expression level and improving the expression level

Active Publication Date: 2022-07-29
ALLIFE MEDICAL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented new way to promote conversion from insulin-producing beta cell precursors (islets) into glucose producing tissue has been developed through an inductive process called induced pluripotentiation or iPSC technology. It uses specialized reagents made up mostly of specific compounds designed specifically target certain genes involved in sugar metabolism. By replacing this old technique used during previous methods like immobilization techniques, researchers have discovered more effective ways to stimulate production of these important proteins such as insulinotropic polypeptide-1(IPP1) and gamma amyloid peptide-42(GAP2). These changes could potentially lead to treating diseases associated with excess levels of blood sugars including Type 2 Diabetes mellitus.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the efficiency and effectiveness of treatments against both types of diabetic conditions such as noninsular carboiditis and Type I diabeteemia. Current approaches involve either injected insulators like artificially produced insulinfusion solutions or autologously controlled insultive transfer systems involving isolated human islets, which may lead to delayed healing and lack of tolerance. There remains a challenge in achieving stable supply of functional pigmentary body cells suitable for use in therapeutic applications without causing adverse side effects associated with current techniques.

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  • Induction method for promoting conversion of pancreas islet alpha cells into beta cells and application of induction method
  • Induction method for promoting conversion of pancreas islet alpha cells into beta cells and application of induction method
  • Induction method for promoting conversion of pancreas islet alpha cells into beta cells and application of induction method

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Embodiment 1

[0187] Example 1 Method for inducing differentiation of iPSCs into islets

[0188] 1. Experimental materials

[0189] The experimental materials involved in the examples of the present invention are shown in Table 1.

[0190] Table 1 Experimental materials

[0191]

[0192]

[0193] 2. Methods of inducing iPSCs to differentiate into islet β cells

[0194] Human-derived iPSCs (from Beijing Chengnuo Medical Technology Co., Ltd., prepared according to the method described in the company's previous patent application (201910110768.7)) with a degree of polymerization of 70%-80% were washed with DPBS phosphate buffer solution 2 times, followed by digestion with TrypLE Express digestion enzyme for 3-5 min at 37°C. After neutralization with DMEM / F12 medium at a ratio of 5:1, centrifugation was performed at 200 g for 5 min. Resuspend the seed with E8 complete medium + 10 μM Y-27632 (ROCK inhibitor) at a density of about 1.8-2.2×10 5 cells / cm 2 , 12-well cell plates were coa...

Embodiment 2

[0227] Example 2 qPCR detection of islet maturation marker gene mRNA expression

[0228] 1. Experimental method

[0229] After iPSCs were induced, Insulin (INS), Glucagon (GCG) and MAFA were used as islet markers to detect the expression of islet marker mRNA in islet cells induced and differentiated by iPSCs.

[0230] Among them, Insulin (INS) refers to insulin, a polypeptide hormone composed of α and β double chains secreted by pancreatic β cells. It consists of 51 amino acids and has a molecular weight of about 5800 Da. Insulin is the only hormone in the body that lowers blood sugar. It is a marker gene of pancreatic beta cells;

[0231] Glucagon (GCG) refers to glucagon, a linear polypeptide hormone composed of 29 amino acids secreted by pancreatic islet alpha cells, with a molecular weight of 3485Da. Has the effect of raising blood sugar in the body;

[0232] MAFA stands for v-maf musculoaponeuroticfibrosarcoma oncogene homologue A, which is a transcription factor with...

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Abstract

The invention discloses an induction method for promoting conversion of islet alpha cells into beta cells and application thereof, and the induction method is characterized in that on the basis of a traditional induction method, on the 35th day of induced differentiation, a culture medium for converting the islet alpha cells into the beta cells is replaced, and meanwhile artemether and a GABAA activator NS11394 are added for induction until the 40th day. Through verification, the induction method disclosed by the invention can be used for remarkably improving the expression quantity of the pancreatic beta cells, and is suitable for large-scale production and clinical application of cell preparations.

Description

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Claims

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Application Information

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Owner ALLIFE MEDICAL SCI & TECH CO LTD
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