Culture additive for in-vitro induction of stem cells to insulin-secretion cells differentiation maturation and purpose thereof

A technology of insulin secretion and cell differentiation, applied in the field of stem cells, can solve the problems of low differentiation efficiency and low cell maturity, and achieve the effect of high induction differentiation efficiency and good cell function maturity.

Inactive Publication Date: 2018-03-16
CHINA JAPAN FRIENDSHIP HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are still problems in the process of induction and differentiation of stem cells into insulin-secreting cells, such as low differentiation efficiency and low cell maturity after differentiation.

Method used

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  • Culture additive for in-vitro induction of stem cells to insulin-secretion cells differentiation maturation and purpose thereof
  • Culture additive for in-vitro induction of stem cells to insulin-secretion cells differentiation maturation and purpose thereof
  • Culture additive for in-vitro induction of stem cells to insulin-secretion cells differentiation maturation and purpose thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Inducing the differentiation of pancreatic adult stem cells into insulin-secreting cells

[0022] Adult pancreatic stem cells (the source and isolation and culture steps can be obtained from the content disclosed in the prior art, and can also be referred to Chinese Journal of Diabetes, 2007, 15(3): 171-175). The isolated and cultured adult pancreatic stem cells were divided into 3×10 5 The density was inoculated in T25 cell culture flasks, using M199 medium, adding 10% fetal bovine serum, and culturing supplement [GLP-1 (final concentration 10 -8 M), nicotinamide (final concentration 5×10 -3 M), 1,25-(OH) 2 D. 3 (final concentration 10 -8 M)]. The culture medium was changed every 3 days, and the cells were subcultured at a ratio of 1:3 after growing to confluency. Taking the M199 medium culture group containing 10% fetal bovine serum without adding the culture supplement of the present invention as a control, the culture was continuously cultured for 4 ...

Embodiment 2

[0023] Example 2 Inducing the differentiation of iPS cells into insulin-secreting cells

[0024] For the establishment steps of iPS cells, please refer to the reference (Cell Res. 2009, 19(4): 429-438). The established iPS cells were divided into 3×10 5 Inoculated in T25 cell culture flasks at a density of 10% fetal calf serum, using M199 medium, culture supplement [Exenatide (final concentration 10 -7 M), nicotinamide (final concentration 1×10 -2 M), 1,25-(OH) 2 D. 3 (final concentration 10 - 7 M)]. The culture medium was changed every 3 days, and the cells were subcultured at a ratio of 1:3 after growing to confluency. Taking the M199 medium culture group containing 10% fetal bovine serum without adding the culture supplement of the present invention as a control, the culture was continuously cultured for 4 weeks. Using standard techniques in the art, real-time PCR was used to detect insulin expression, and ELISA was used to detect glucose-stimulated insulin release....

Embodiment 3

[0025] Example 3 Inducing the differentiation of liver tissue-derived stem cells into insulin-secreting cells

[0026] Stem cells were isolated from liver tissue using published techniques. Hepatic stem cells were divided into 3×10 5 Inoculate in the T25 cell culture flask with the density of M199 medium, add 10% fetal bovine serum, culture supplement [liraglutide (final concentration 10 -9 M), nicotinamide (final concentration 2×10 -3 M), 1,25-(OH) 2 D. 3 (final concentration 10 -9 M)]. The culture medium was changed every 3 days, and the cells were subcultured at a ratio of 1:3 after growing to confluency. Taking the M199 medium culture group containing 10% fetal bovine serum without adding the culture supplement of the present invention as a control, the culture was continuously cultured for 4 weeks. Using standard techniques in the art, real-time PCR was used to detect insulin expression, and ELISA was used to detect glucose-stimulated insulin release. The result i...

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Abstract

The invention discloses a culture supplement for inducing differentiation and maturation of stem cells to insulin-secreting cells in vitro and its use in inducing differentiation of stem cells. The supplement can promote the differentiation of stem cells from various sources into insulin-secreting cells in vitro, including adult stem cells derived from pancreatic tissue, induced pluripotent stem cells (iPS cells) or stem cells derived from other tissues. The main components of the culture supplement of the present invention are glucagon-like peptide 1 or its analogue, nicotinamide, and 1,25‑(OH)2D3. The culture supplement can be used in conjunction with any basal medium to induce stem cells to differentiate into insulin-secreting cells with good efficiency and the maturity of differentiated cells. The culture supplement disclosed by the invention can be used to induce differentiation of stem cells into insulin-secreting cells, so as to prepare stem cell products that can replace the function of pancreatic islets.

Description

technical field [0001] The invention relates to the technical field of stem cells, in particular to a culture supplement for inducing differentiation of adult stem cells or induced pluripotent stem cells (induced pluripotent stem cells, iPS) into insulin-secreting cells in vitro. And the application of the additive in the differentiation of stem cells to insulin-secreting cells. Background technique [0002] Diabetes is expected to become a major disease that threatens human health with high morbidity and serious complications. At present, the use of hypoglycemic drugs or insulin therapy can improve the patient's glucose metabolism disorder to a certain extent, but it cannot effectively prevent or reverse the microvascular disease caused by diabetes. disease and its complications. Pancreatic islet transplantation is an effective method for treating insulin-dependent diabetes mellitus, and experimental data show that islet transplantation can not only correct the disorder of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/074
CPCC12N5/0676C12N5/0602C12N2500/35C12N2500/46C12N2501/335C12N2506/03C12N2506/45
Inventor 娄晋宁张文健
Owner CHINA JAPAN FRIENDSHIP HOSPITAL
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