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Method for the differentiation of human adult stem cells into insulin-secreting cells

A technology of adult stem cells and insulin secretion, applied in the directions of insulin, chemical instruments and methods, biochemical equipment and methods, etc., can solve problems such as difficulty in using embryonic stem cells to treat diabetes

Inactive Publication Date: 2010-06-09
孔喜淑
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As mentioned above, racial issues and cancer development make it difficult to use embryonic stem cells to treat diabetes

Method used

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  • Method for the differentiation of human adult stem cells into insulin-secreting cells
  • Method for the differentiation of human adult stem cells into insulin-secreting cells
  • Method for the differentiation of human adult stem cells into insulin-secreting cells

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Experimental program
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Effect test

Embodiment 1

[0039] Example 1 Separation of Stem Cells

[0040] The adipose tissue surgically dissected from the subcutaneous layer around the eyes in a plastic surgery hospital was treated with 0.075% type 1 collagenase solution at 37°C for 30 min, and then supplemented with the same volume of collagenase solution with Medium with fetal bovine serum terminates enzyme activity. After centrifugation, the cell pellet thus formed was washed twice with medium and incubated in DMEM-LG supplemented with 10% fetal bovine serum.

Embodiment 2

[0041] Embodiment 2 stem cell culture

[0042] The obtained stem cells were cultured for 21 days in a differentiation induction medium. For this, first, the cells were divided into 5 × 10 3 Cells / well density were seeded in collagen-coated 48-well plates to which was added 25 mM glucose supplemented with 10% fetal calf serum, 4 nM activin prior to incubation (for 7 days). A. Medium (DMEM-HG) with 10 nM glucagon-like peptide-1 and 20 ng / ml fibroblast growth factor-2. Thereafter, the cells were incubated with 5.5 mM glucose supplemented with 10% fetal bovine serum, 10 mM nicotinamide, 4 nM activin A, 10 nM glucagon-like peptide-1, and 50 ng / ml insulin-like growth factor-1 or insulin-like Growth factor-2 medium (DMEM-LG) was further cultured for another 14 days.

[0043] During the incubation period, the medium was replaced with fresh medium every 3-4 days. After a total of 3 weeks of incubation, cells were examined for differentiation as follows.

Embodiment 3

[0044] Morphological changes of cells after differentiation in embodiment 3

[0045] Stem cells were isolated from the subcutaneous fat around the eyes and cultured to monitor their morphological changes. Although morphologically unchanged after incubation in growth factor and cytokine-free media, when they were treated with growth factors and / or cytokines such as nicotinamide, activin A and glucagon-like peptide-1 or When treated with IGF-1 or -2, it was observed that the stem cells underwent a morphological change into a round shape ( figure 1 ). After 3 weeks of differentiation induction, the cells became rounded in morphology and formed cell clusters.

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Abstract

Disclosed is a method for differentiating human adult stem cells into insulin-secreting cells. Human adult stem cells, isolated from the subcutaneous adipose tissues around the eyes, are induced to differentiate into insulin-secreting cells in a medium in the presence of cytokines and growth factors including B27 supplement, fibroblast growth factor-2, epidermal growth factor, nicotinamide, glucagon-like peptide-1, activin A, insulin-like growth factor, betacellulin, etc., with a glucose shift from a high concentration to a low concentration. Having the ability to producing insulin and C-peptide at high levels, the insulin-secreting cells can be excellent curatives for type 1 diabetes mellitus.

Description

technical field [0001] The present invention relates to inducing the differentiation of human adult stem cells into insulin-secreting cells. Background technique [0002] Type 1 diabetes, also known as insulin-dependent diabetes, is characterized by the loss of insulin-producing beta cells, usually resulting from an autoimmune attack of the islets in the pancreas, resulting in a deficiency of insulin. Insulin plays a key role in the homeostasis of blood sugar levels in the body. When insulin is deficient or produced in low amounts, blood sugar levels increase, leading to complications in various organs such as kidneys, nerves, retina, and the like. Treatment of type 1 diabetes relies mostly on the use of insulin, which needs to be injected throughout the patient's lifetime. Insulin use is not a fundamental treatment for type 1 diabetes. On the other hand, transplantation of exogenous pancreas or beta cells is used to treat type 1 diabetes. However, this method suffers fr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08A61K35/12A61K35/39C12N5/07C12N15/09C12Q1/68
CPCC12N2533/54C12N2501/335C12N5/0676C12N2501/16C12N2501/115C12N2501/105C12N2500/38C12N2506/1384C12N2500/34A61P3/10C12N5/0602C12N5/00C07K14/62
Inventor 金海权姜贤美
Owner 孔喜淑
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