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Method and combined medium for preparing insulin-like secreting cells

An insulin-like, culture medium technology, applied in the fields of biology and medicine, can solve the ethical limitations of embryonic stem cells, immune rejection of allogeneic transplantation, limited resources, etc., and achieve the effects of prolonging survival time, improving diabetes symptoms, and increasing body weight.

Inactive Publication Date: 2009-11-04
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] It has been reported that mouse and human embryonic stem cells can be induced to differentiate into insulin-secreting cells, but embryonic stem cells have ethical restrictions and limited resources, and allograft immune rejection still exists after transplantation

Method used

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  • Method and combined medium for preparing insulin-like secreting cells
  • Method and combined medium for preparing insulin-like secreting cells
  • Method and combined medium for preparing insulin-like secreting cells

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Embodiment 1. A combined medium for preparing insulin-like secreting cells, which consists of independent medium I, medium II and medium III;

[0048] Medium I is: DMEM high-glucose medium, 1% FBS, 10ng / ml bFGF, 1% DMSO, 23mmol / l glucose;

[0049] Medium II is: serum-free DMEM / F12 medium, 17.5mmol / l glucose, 10mmol / l nicotinamide, 20ng / ml EGF, 20ng / ml bFGF, 10nmol / l exendin-4, 1% B27 supplement and 1% N2 additive;

[0050] Medium III is: RPMI 1640 medium, 11.1mmol / l glucose, 10mmol / l nicotinamide, 10mmol / l Hepes, 100pmol / l HGF, 2nmol / l activin-A, 10nmol / l exendin-4.

[0051] The production methods of medium I, medium II and medium III are all conventional mixing; medium I, medium II and medium III can be stored independently at 4°C, and the effective storage period is 30 days.

Embodiment 2

[0052] Embodiment 2. A combined medium for preparing insulin-like secreting cells, which consists of independent medium I, medium II and medium III;

[0053] Medium I is: DMEM high-glucose medium, 1.5% FBS, 11ng / ml bFGF, 1% DMSO, 25mmol / l glucose;

[0054] Medium II is: serum-free DMEM / F12 medium, 19mmol / l glucose, 11mmol / l nicotinamide, 22ng / mlEGF, 22ng / ml bFGF, 10nmol / l exendin-4, 1% B27 additive and 1% N2 additive;

[0055] Medium III is: RPMI 1640 medium, 12mmol / l glucose, 11mmol / l nicotinamide, 11mmol / l Hepes, 110pmol / l HGF, 2.5nmol / l activin-A, 11nmol / l exendin-4.

[0056] The production method is the same as above.

Embodiment 3

[0057] Embodiment 3. A combined medium for preparing insulin-like secreting cells, consisting of independent medium I, medium II and medium III;

[0058] Medium I is: DMEM high-glucose medium, 2% FBS, 9ng / ml bFGF, 1% DMSO, 17mmol / l glucose;

[0059] Medium II is: serum-free DMEM / F12 medium, 15mmol / l glucose, 9mmol / l nicotinamide, 18ng / mlEGF, 18ng / ml bFGF, 10nmol / l exendin-4, 1% B27 additive and 1% N2 additive;

[0060] Medium III is: RPMI 1640 medium, 10mmol / l glucose, 9mmol / l nicotinamide, 9mmol / l Hepes, 90pmol / l HGF, 1.5nmol / l activin-A, 9nmol / l exendin-4.

[0061] The production method is the same as above.

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Abstract

The invention discloses a combined medium for preparing insulin-like secreting cells, which consists of a medium I, a medium II and a medium III which are independent respectively, wherein, the medium I is a DMEM (Dulbecco's modified Eagle's medium) high-glucose medium prepared from fetal calf sera, basic fibroblast growth factors (bFGF), dimethyl sulfoxide (DMSO) and glucose; the medium II is a serum-free DMEM / F12 medium prepared from glucose, niacinamide, epidermal growth factors (EGF), bFGF, exendin-4, a B27 additive and an N2 additive; and the medium III is an RPMI (Roswell Park Memorial Institute) 1640 medium prepared from glucose, niacinamide, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, hepatocyte growth factors (HGF), activin A and exendin-4. The invention further discloses a method using the combined medium for preparing insulin-like secreting cells at the same time. The invention can facilitate the differentiation of hBM-MSC (human bone marrow mesenchymal stem cell) into insulin-like secreting cells having the effect of correcting diabetes hyperglycemia.

Description

technical field [0001] The invention belongs to the technical field of biology and medicine, in particular to a method for preparing insulin-like secreting cells by using adult bone marrow mesenchymal stem cells (hBM-MSCs) and the used combination culture medium. Background technique [0002] With the improvement of living standards, diabetes has become a major social problem that threatens the health and life of all human beings, and has caused a huge economic burden to the society. Pancreatic islet transplantation is considered to be the most effective way to treat insulin-dependent diabetes mellitus. However, the widespread clinical development of islet transplantation is limited due to the severe shortage of donor pancreas. Allogeneic transplantation cannot be really used in clinical practice due to serious immune rejection problems. In recent years, the development of stem cell technology and the regulation of stem cell islet differentiation technology have become a ne...

Claims

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Application Information

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IPC IPC(8): C12N5/08
Inventor 吴育连谢秋萍
Owner ZHEJIANG UNIV
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