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Islet cells differentiated from stem cells, method, compound and application

A technology of stem cell differentiation and islet cells, applied in the field of islet cells, can solve the problems of increased tumorigenic risk and low efficiency, and achieve the effect of good diabetes, high efficiency and good treatment

Pending Publication Date: 2021-07-27
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Transplanting undifferentiated adipose-derived mesenchymal stem cells into the body may increase the risk of tumorigenesis, and the efficiency of the induced insulin-secreting cells is currently low, so the effect of treating diabetes still needs to be further improved

Method used

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  • Islet cells differentiated from stem cells, method, compound and application
  • Islet cells differentiated from stem cells, method, compound and application
  • Islet cells differentiated from stem cells, method, compound and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] This embodiment provides a method for stem cells to differentiate into islet cells, comprising the following steps:

[0043] (1) Extraction and separation to obtain high-purity adipose-derived mesenchymal stem cells with differentiation ability;

[0044] (2) Take adipose-derived mesenchymal stem cells (ADSCs), and culture them in a six-well plate with a cell number of 4×105 / well. When the confluence of the cells is about 90%, discard the medium I and add Adv -PDX-1 overexpressed adenovirus medium infected cells 6h after the replacement of fresh medium I, and placed in an incubator to continue culturing for 7d, medium I includes: culture medium (L-DMEM) + 10% North American fetal bovine serum (Gibco FBS), Adv-PDX-1 is a recombinant adenovirus inserted into the CDS region of the rat PDX-1 gene, and the sequence of the CDS region is shown in Sequence Table 1. After 7d, such as figure 1 As shown, after adipose-derived mesenchymal stem cells (ADSCs) were infected with Adv-...

Embodiment 2

[0051] The difference with embodiment 1 is:

[0052] In step (2), when the confluence of the cells is about 85%, the medium I is discarded, and the medium containing Adv-PDX-1 overexpressed adenovirus is added to infect the cells for 5 hours, and then fresh medium I is replaced, and placed in the culture medium. Continue to cultivate in the box for 8 days;

[0053] In step (3), add induction medium I and continue culturing for 4 days, during which induction medium I is replaced every 48h;

[0054] In step (4), add induction medium II and continue to cultivate for 6 days, during which the induction medium II is replaced every 40 h;

[0055] In step (5), add the induction medium III and continue to cultivate for 6 days, during which the induction medium III is replaced every 40 hours, and the target cell clusters are obtained after 6 days and collected for storage;

[0056] The culture conditions are: at 35°C, 96% humidity in CO 2 cultured in an incubator.

Embodiment 3

[0058] The difference with embodiment 1 is:

[0059] In step (2), when the confluence of the cells is about 95%, the medium I is discarded, and the medium containing Adv-PDX-1 overexpressed adenovirus is added to infect the cells for 7 hours, and then fresh medium I is replaced, and placed in the culture medium. Continue to cultivate in the box for 6 days;

[0060] In step (3), add induction medium I and continue culturing for 6 days, during which induction medium I is replaced every 40 h;

[0061] In step (4), add induction medium II and continue to cultivate for 8 days, during which the induction medium II is replaced every 48h;

[0062] In step (5), add the induction medium III and continue culturing for 8 days, during which the induction medium III is replaced every 48 hours, and the target cell clusters are obtained after 8 days and collected and stored;

[0063] The culture conditions are: at 36°C, 93% humidity in CO 2 cultured in an incubator.

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Abstract

The invention is applicable to the technical field of biomedicine, and provides islet cells differentiated from stem cells, a method, a compound and application. The method for differentiating the stem cells into the islet cells comprises the following steps: conducting extracting and separating to obtain high-purity adipose-derived mesenchymal stem cells with differentiation capacity; transferring the PDX-1 gene into the adipose-derived mesenchymal stem cells by adopting an adenovirus infection method; and directionally differentiating the adipose-derived stem cells into islet cells by using an inducer. The adipose-derived mesenchymal stem cells are directionally differentiated into the islet cells by combining an adenovirus infection method with an inducer, the insulin (INS) recombinant protein is highly expressed in the obtained islet cell cluster, and the PDX-1 protein is still highly expressed in most cells after induction is finished, so that the induced cells have the capability of secreting the insulin (INS) recombinant protein and are high in efficiency; and an islet cell carrying cytoskeleton compound is transplanted to a skeletal muscle part, and diabetes can be well treated.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an islet cell differentiated from stem cells, a method, a compound and an application. Background technique [0002] With the continuous improvement of people's living standards, the global incidence of diabetes is increasing year by year. Diabetes is mainly characterized by relative or absolute lack of insulin, accompanied by chronic metabolic diseases characterized by persistent hyperglycemia. In addition, it may also cause a series of complications, such as diabetic nephropathy and diabetic macrovascular disease. The quality of life of the people has also brought enormous pressure to the national economy. At present, the treatment of diabetes mainly relies on insulin, oral hypoglycemic drugs, or allogeneic islet transplantation. Either way, there are certain hidden dangers and inconveniences. For patients, islet transplantation is more likely to have economic,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/861C12N15/12C12N5/10A61K35/39A61P3/10
CPCC12N15/86C07K14/4705C12N5/0667C12N5/0676A61K35/39A61P3/10C12N2710/10343C12N2800/107C12N2510/00C12N2506/1384C12N2501/115C12N2501/11C12N2501/998C12N2500/25C12N2501/12C12N2501/16C12N2500/33C12N2501/33C12N2533/74C12N2533/40C12N2533/30
Inventor 陈立张明张文友邵丹杨雪晗
Owner JILIN UNIV
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