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Culture medium and application thereof and method for preparing mesenchyme stem cells through urine cells

A technology of stem cells and urine cells, applied in the field of stem cells and genetic engineering, can solve the problems of low immunity, immune response to the human body, decline in proliferation and differentiation ability, etc., and achieve the effect of wide source, high activity rate and large quantity

Active Publication Date: 2017-08-22
GUANGZHOU RAINHOME PHARM&TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the main source of mesenchymal stem cells is bone marrow and umbilical cord tissue. As bone marrow mesenchymal stem cells age, the number of stem cells decreases significantly, and the ability to proliferate and differentiate significantly declines. Immune response and greater damage to the human body
Although umbilical cord mesenchymal stem cells have low immunity, the preservation of umbilical cord blood is more expensive
The number of stem cells from other tissue sources is very rare, and it is also very difficult to isolate and culture

Method used

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  • Culture medium and application thereof and method for preparing mesenchyme stem cells through urine cells
  • Culture medium and application thereof and method for preparing mesenchyme stem cells through urine cells
  • Culture medium and application thereof and method for preparing mesenchyme stem cells through urine cells

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] 1. Preparation of culture medium

[0054] IPS induction medium (configuration medium A): REGM and MEF medium were mixed in a volume ratio (1:1).

[0055] Embryoid body induction medium (configuration medium B): 450mL DMEM, 50mL FBS, 5mML-glutamine, 0.1mM β-mercaptoethanol, 50ng / mL BMP-4, 25ng / mLVEGF, 60pM insulin.

[0056] Embryoid body medium (configuration medium C): 400mLDMEM, 100mL FBS, 5ug / ml insulin transferrin sodium selenite (ITS), 0.23mM sodium pyruvate, 1ng / ml epidermal growth factor (EGF), 2mML- Glutamine, 0.1 mM β-mercaptoethanol.

[0057] Mesenchymal stem cell induction medium (configuration medium D): 450mL DMEM basal medium, 50mL FBS, 60pM insulin, 2mML-glutamine, 75ug / LbFGF, 3×10-8mol / L dexamethasone.

[0058] Mesenchymal stem cell medium (configuration medium E): 450mLDMEM basal medium, 50mLFBS, 10ng / mLhEGF, 10ng / mL bFGF, 3ng / mL HGF, 5ng / mLPDGF, 5ng / mLTGF-β.

[0059] 2. Acquisition of urine cells

[0060] 1) Add 2mL penicillin / streptomycin double an...

Embodiment 2

[0085] 1. Preparation of culture medium

[0086] IPS induction medium (configuration medium A): REGM and MEF medium were mixed in a volume ratio (1:1).

[0087] Embryoid body induction medium (configuration medium B): 450mL MEM, 50mL FBS, 1mM-glutamine, 0.05mM β-mercaptoethanol, 10ng / mL BMP-4, 1ng / mLVEGF, 1pM insulin.

[0088] Embryoid body medium (configuration medium C): 400mLDMEM, 100mL FBS, 1ug / ml insulin transferrin sodium selenite (ITS), 0.1mM sodium pyruvate, 0.2ng / ml epidermal growth factor (EGF), 1mML - Glutamine, 0.01 mM β-mercaptoethanol.

[0089] Mesenchymal stem cell induction medium (configuration medium D): 450mL DMEM basal medium, 50mL FBS, 1pM insulin, 1mML-glutamine, 50ug / LbFGF, 2×10-8mol / L dexamethasone.

[0090] Mesenchymal stem cell medium (configuration medium E): 450mLDMEM basal medium, 50mLFBS, 1ng / mLhEGF, 1ng / mL HGF, 1ng / mL bFGF, 2ng / mLPDGF, 1ng / mLTGF-β.

[0091] 2. Acquisition of urine cells

[0092] 1) Add 2mL penicillin / streptomycin double antib...

Embodiment 3

[0117] 1. Preparation of culture medium

[0118] IPS induction medium (configuration medium A): REGM and MEF medium were mixed in a volume ratio (1:1).

[0119] Embryoid body induction medium (configuration medium B): 450mL MEM, 50mL FBS, 10mM-glutamine, 0.5mM β-mercaptoethanol, 85ng / mL BMP-4, 50ng / mLVEGF, 100pM insulin.

[0120] Embryoid body medium (configuration medium C): 400mLDMEM, 100mL FBS, 10ug / ml insulin transferrin sodium selenite (ITS), 0.65mM sodium pyruvate, 5ng / ml epidermal growth factor (EGF), 5mM L - Glutamine, 100 mM β-mercaptoethanol.

[0121] Mesenchymal stem cell induction medium (configuration medium D): 450mL DMEM basal medium, 50mL FBS, 100pM insulin, 10mML-glutamine, 150ug / LbFGF, 5×10 -8 mol / L dexamethasone.

[0122] Mesenchymal stem cell medium (configuration medium E): 450mLDMEM basal medium, 50mLFBS, 100ng / mLhEGF, 100ng / mL bFGF, 50ng / mL HGF, 20ng / mLPDGF, 25ng / mLTGF-β.

[0123] 2. Acquisition of urine cells

[0124] 1) Add 2mL penicillin / streptom...

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Abstract

The invention relates to the technical field of stem cells and genetic engineering, in particular to a culture medium and application thereof and a method for preparing mesenchyme stem cells through urine cells. According to the method capable of inducing the urine cells to become the mesenchyme stem cells in the culture medium, the urine cells are wide in source, and are not limited by ethic, and through the adoption of the method, the urine cells form IPS cells embryoid bodies-mesenchyme stem cells in sequence. All induction can be completed in 20-25 days, and the obtained cells conform to characteristics of the mesenchyme stem cells after being detected, are large in quantity, and high in motility rate.

Description

technical field [0001] The invention relates to the technical field of stem cells and genetic engineering, in particular to a culture medium and its application and a preparation method of mesenchymal stem cells. Background technique [0002] With the rise of tissue engineering and clinical transplantation medicine, the research on seed cells has attracted more and more attention. Mesenchymal stem cells (MSC, mesenchymal stem cells) are a type of pluripotent stem cells that exist in various tissues (such as bone marrow, umbilical cord blood and umbilical cord tissue, placental tissue, adipose tissue, etc.). It has the ability of high proliferation and self-renewal, and has the potential of multi-lineage differentiation. It can be differentiated into cartilage, bone, skeletal muscle, tendon, fat, dermis, nerve and kidney parenchymal cells under different induction conditions. Therefore, mesenchymal stem cells are often used as seed cells. [0003] Clinical research on mese...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N5/0775
CPCC12N5/0662C12N2500/25C12N2500/32C12N2500/44C12N2501/11C12N2501/115C12N2501/12C12N2501/135C12N2501/15C12N2501/155C12N2501/165C12N2501/39C12N2501/602C12N2501/603C12N2501/604C12N2501/605C12N2501/608C12N2506/25C12N2506/45C12N2510/00C12N2533/54
Inventor 黄燕飞车七石赵澎刘少辉
Owner GUANGZHOU RAINHOME PHARM&TECH CO LTD
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