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Culture medium and culture method for inducing multipotentstem cells for directional differentiation of renal cells

A technology for pluripotent stem cells and directed differentiation, which is applied in the field of culture medium for inducing pluripotent stem cells directed differentiation of kidney cells. The effect of differentiation

Active Publication Date: 2017-08-18
GUANGZHOU RAINHOME PHARM&TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the medium for inducing human IPS cells to differentiate into kidney cells in vitro has a low induction rate, and most of the medium contains fetal bovine serum, which cannot meet the large needs of research and production.

Method used

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  • Culture medium and culture method for inducing multipotentstem cells for directional differentiation of renal cells
  • Culture medium and culture method for inducing multipotentstem cells for directional differentiation of renal cells
  • Culture medium and culture method for inducing multipotentstem cells for directional differentiation of renal cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 A culture medium for the directed differentiation of induced pluripotent stem cells into kidney cells

[0029]A medium for induced pluripotent stem cells to differentiate into kidney cells, consisting of the following components and their concentrations: DEME basal medium 12g / L, mannitol 0.45mM, heparin sodium 5U, L-glutamine 1.25mM, sodium pyruvate 0.68mM, β-mercaptoethanol 0.17mM, fibronectin 2.4mg / L, directed differentiation induction factor 36ng / mL and serum replacement.

[0030] The directional differentiation inducing factor is composed of the following components and concentrations: activin A 12ng / mL, human vascular endothelial growth factor 8.2ng / mL, β-glycerol phosphate 6.8ng / mL and tanshinone ⅡA 9ng / mL.

[0031] The serum substitute is composed of the following components and their concentrations: oat protein 7.5g / L, transferrin 0.45g / L, recombinant human epidermal growth factor 1.2ng / mL, fibroblast growth factor 1.75ng / mL, platelet source Growth fa...

Embodiment 2

[0037] Example 2 A culture medium for the directed differentiation of induced pluripotent stem cells into kidney cells

[0038] A medium for induced pluripotent stem cells to differentiate into kidney cells, consisting of the following components and their concentrations: DEME basal medium 10g / L, mannitol 0.1mM, heparin sodium 4U, L-glutamine 1mM, sodium pyruvate 0.5 mM, β-mercaptoethanol 0.15mM, fibronectin 1mg / L, directed differentiation induction factor 27ng / mL and serum replacement.

[0039] The directional differentiation inducing factor is composed of the following components and concentrations: activin A 11ng / mL, human vascular endothelial growth factor 7.5ng / mL, β-phosphoglycerol 3.5ng / mL and tanshinone ⅡA 5ng / mL.

[0040] The serum substitute consists of the following components and their concentrations: avenous protein 5g / L, transferrin 0.3g / L, recombinant human epidermal growth factor 0.5ng / mL, fibroblast growth factor 1ng / mL, platelet-derived growth Factor 0.75ng / ...

Embodiment 3

[0042] Example 3 A medium for the directed differentiation of induced pluripotent stem cells into kidney cells

[0043] A medium for induced pluripotent stem cells to differentiate into kidney cells, consisting of the following components and their concentrations: DEME basal medium 15g / L, mannitol 0.5mM, heparin sodium 8U, L-glutamine 1.5mM, sodium pyruvate 0.85mM, β-mercaptoethanol 0.2mM, fibronectin 3mg / L, directed differentiation induction factor 39ng / mL and serum replacement.

[0044] The directional differentiation inducing factor is composed of the following components and concentrations: activin A 12.5ng / mL, human vascular endothelial growth factor 9ng / mL, β-phosphoglycerol 7.5ng / mL and tanshinone ⅡA 10ng / mL.

[0045] The serum substitute consists of the following components and their concentrations: avenous protein 10g / L, transferrin 0.5g / L, recombinant human epidermal growth factor 1.5ng / mL, fibroblast growth factor 2ng / mL, platelet-derived growth Factor 2.5ng / mL, gl...

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Abstract

The invention relates to the field of cell engineering, and in particular relates to a culture medium and a culture method for inducing multipotent stem cells for directional differentiation of renal cells. The culture medium for inducing the multipotent stem cells for directional differentiation of the renal cellscomprises 10-15g / L of a DEME basal medium, 0.1-0.5mM of mannitol, 4-8U of heparin sodium, 1-1.5mM of L-glutamine, 0.5-0.85mM of sodium pyruvate, 0.15-0.2mM of beta-mercaptoethanol, 1-3mg / L of fibronectin, 27-39ng / mL of directional differentiation induction factors, and a serum replacement. The culture medium for inducing the multipotent stem cells for directional differentiation of the renal cells, provided by the invention, can effectively promote induction of the directional differentiation from the multipotent stem cells to the renal cells, and no fetal calf serum is added in the culture medium, thereby avoiding a risk of infection of animal pathogens.

Description

technical field [0001] The invention relates to the field of cell engineering, in particular to a culture medium and a culture method for inducing pluripotent stem cells to differentiate kidney cells. Background technique [0002] Chronic kidney disease is one of the common clinical diseases. According to the latest epidemiological survey, there are currently 120 million chronic kidney disease patients in my country, with a prevalence rate as high as 10.8%, and it is showing a growing trend year by year. Kidney transplantation is the most effective clinical treatment, but there are problems such as shortage of donors, allogeneic rejection and life-long administration of immunosuppressants. Therefore, finding seed cells with strong regeneration ability and no rejection becomes the key to solve this problem. At present, the cells used in regenerative medicine research mainly include adult stem cells and embryonic stem cells, but these two types of cells have certain disadvan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0686C12N2500/24C12N2500/30C12N2500/32C12N2500/35C12N2500/38C12N2500/44C12N2501/11C12N2501/113C12N2501/115C12N2501/119C12N2501/91C12N2501/998C12N2506/45
Inventor 车七石
Owner GUANGZHOU RAINHOME PHARM&TECH CO LTD
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