Method for inducing pancreas islet cells differentiated by stem cells in vitro to form pancreas islet-like structure

A technology for stem cell differentiation and islet cells, applied in cell dissociation methods, artificial cell constructs, biochemical equipment and methods, etc., can solve problems such as low induction rate and negative impact on product quality, and achieve high induction rate and application effect Good, the effect of high product cell quality

Active Publication Date: 2021-11-02
多能干细胞再生医学科技(广州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of low induction rate and negative impact on product quality in the above-mentioned existing methods, the present invention provides a new method capable of rapidly and efficiently inducing stem cell differentiation in vitro to form islet-like structures. The induction rate is high, The product quality is high, and the specific plan is as follows:

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 In vitro induction of stem cell-differentiated islet cells to form islet-like structures:

[0031] S1. Obtaining mesenchymal stem cells by separating and culturing from raw material tissues;

[0032] S2, subculture of the obtained mesenchymal stem cells;

[0033] S3. Inducing and differentiating the obtained passaged mesenchymal stem cells into islet-like cells.

[0034] The raw material tissue in S1 includes human placenta tissue.

[0035] The isolation culture described in S1 includes: washing the raw material tissue, cutting it into pieces, digesting it with digestive enzymes, centrifuging to obtain a cell pellet, and inoculating it in a cell culture medium after washing.

[0036] The above-mentioned digestive enzymes include type I collagenase, and the digestion time is 45 minutes.

[0037] For the above centrifugation, the condition is to centrifuge at 1200 rpm for 5 minutes; for the above washing, use PBS to wash 3 times.

[0038] After being inocula...

Embodiment 2

[0049] Example 2 In vitro induction of stem cell-differentiated islet cells to form islet-like structures:

[0050] S1. Obtaining mesenchymal stem cells by separating and culturing from raw material tissues;

[0051]S2, subculture of the obtained mesenchymal stem cells;

[0052] S3. Inducing and differentiating the obtained passaged mesenchymal stem cells into islet-like cells.

[0053] The raw material tissue in S1 includes adipose tissue.

[0054] The isolation culture described in S1 includes: washing the raw material tissue, cutting it into pieces, digesting it with digestive enzymes, centrifuging to obtain a cell pellet, and inoculating it in a cell culture medium after washing.

[0055] The above-mentioned digestive enzymes include type II collagenase, and the digestion time is 60 minutes.

[0056] For the above centrifugation, the condition is to centrifuge at 1500 rpm for 8 minutes; for the above washing, use PBS to wash 5 times.

[0057] After being inoculated in ...

Embodiment 3

[0068] Example 3 In vitro induction of stem cell-differentiated islet cells to form islet-like structures:

[0069] S1. Obtaining mesenchymal stem cells by separating and culturing from raw material tissues;

[0070] S2, subculture of the obtained mesenchymal stem cells;

[0071] S3. Inducing and differentiating the obtained passaged mesenchymal stem cells into islet-like cells.

[0072] The raw material tissue in S1 includes placental tissue.

[0073] The isolation culture described in S1 includes: washing the raw material tissue, cutting it into pieces, digesting it with digestive enzymes, centrifuging to obtain a cell pellet, and inoculating it in a cell culture medium after washing.

[0074] The above-mentioned digestive enzymes include type I collagenase or type II collagenase, and the digestion time is 50 minutes.

[0075] For the above centrifugation, the condition is 1400 rpm for 6 minutes; for the above washing, PBS is used for washing 4 times.

[0076] After bein...

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Abstract

The invention discloses a method for in-vitro induction of stem cell differentiated islet cells to form islet-like structures, and relates to the technical field of biologication. The method comprises the following steps: enabling raw material tissue to be subjected to isolated culture to obtain mesenchymal stem cells; carrying out subculture on the obtained mesenchymal stem cells; and carrying out induced differentiation on the obtained passage mesenchymal stem cells to differentiate into islet-like cells. By adopting the specific induction culture medium and the steps of pretreatment and induction culture, the islet cells differentiated by the stem cells can be rapidly and efficiently induced in vitro to form the islet-like structure, and the method is high in induction rate, high in product cell quality and good in application effect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for inducing stem cell-differentiated islet cells to form islet-like structures in vitro. Background technique [0002] Diabetes is a group of metabolic diseases characterized by hyperglycemia. Hyperglycemia is caused by defective insulin secretion or impaired biological action, or both. Long-term high blood sugar leads to chronic damage and dysfunction of various tissues, especially the eyes, kidneys, heart, blood vessels, and nerves. [0003] Islet transplantation is a fundamental method for the treatment of diabetes that emerged in this century. However, in the early stage, islet isolation and transplantation were mainly used, combined with appropriate immunosuppressive regimens. Such regimens are extremely dependent on donors, so large-scale applications cannot be achieved. In recent years, the application of stem cell differentiation in the medical field has gradually...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/0775
CPCC12N5/0676C12N5/0662C12N5/0668C12N5/0667C12N2506/1346C12N2506/1392C12N2506/1384C12N2509/10C12N2509/00C12N2501/135C12N2501/115C12N2500/38C12N2501/11C12N2501/998
Inventor 顾军顾帅兰丹
Owner 多能干细胞再生医学科技(广州)有限公司
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