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Medium, purpose thereof and method of inducing multipotent stem cells to be transformed to mesenchymal stem cells

A technology of stem cells and cell transformation, applied in the field of stem cells and genetic engineering, can solve the problems of low immunity, immune response to the human body, and the decline of proliferation and differentiation ability, and achieve the effects of wide source, high viability and large quantity

Active Publication Date: 2017-09-08
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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  • Medium, purpose thereof and method of inducing multipotent stem cells to be transformed to mesenchymal stem cells
  • Medium, purpose thereof and method of inducing multipotent stem cells to be transformed to mesenchymal stem cells
  • Medium, purpose thereof and method of inducing multipotent stem cells to be transformed to mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Preparation of culture medium

[0039] IPS induction medium (configuration medium A): REGM and MEF medium were mixed according to volume ratio.

[0040] Mesenchymal stem cell induction medium (configuration medium B): 450mLDMEM basal medium, 50mLFBS, 80pM insulin, 5mM L-glutamine, 85ug / LbFGF, 15ug / LSCF, 3×10 -8 mol / L dexamethasone.

[0041] Mesenchymal stem cell medium (configuration medium C): 450mLDMEM basal medium, 50mLFBS, 10ng / mLhEGF, 10ng / mL bFGF, 3ng / mL HGF, 5ng / mLPDGF, 5ng / mLTGF-β.

[0042] 2. Acquisition of urine cells

[0043] 1) Add 2mL penicillin / streptomycin double antibody to each collection cup.

[0044] 2) Collect the urine. If the follow-up operation is not performed immediately, store the urine in a 4°C refrigerator and complete the follow-up operation on the same day.

[0045] 3) One well of a six-well plate was prepared for each urine, and the well was coated with 0.1% gelatin for more than 20 minutes, and the liquid in the well was blotted be...

Embodiment 2

[0063] 1. Preparation of culture medium

[0064] IPS induction medium (configuration medium A): REGM and MEF medium were mixed according to volume ratio.

[0065] Mesenchymal stem cell induction medium (configuration medium B): 450mLDMEM basal medium, 50mLFBS, 1pM insulin, 1mM L-glutamine, 50ug / LbFGF, 5ug / L SCF, 2×10 -8 mol / L dexamethasone.

[0066] Mesenchymal stem cell medium (configuration medium C): 450mLDMEM basal medium, 50mLFBS, 1ng / mLhEGF, 1ng / mL bFGF, 1ng / mL HGF, 2ng / mLPDGF, 1ng / mLTGF-β.

[0067] 2. Acquisition of urine cells

[0068] 1) Add 2mL penicillin / streptomycin double antibody to each collection cup.

[0069] 2) Collect the urine. If the follow-up operation is not performed immediately, store the urine in a 4°C refrigerator and complete the follow-up operation on the same day.

[0070] 3) One well of a six-well plate was prepared for each urine, and the well was coated with 0.1% gelatin for more than 20 minutes, and the liquid in the well was blotted befor...

Embodiment 3

[0088] 1. Preparation of culture medium

[0089] IPS induction medium (configuration medium A): REGM and MEF medium were mixed according to volume ratio.

[0090] Mesenchymal stem cell induction medium (configuration medium B): 450mLDMEM basal medium, 50mLFBS, 160pM insulin, 10mML-glutamine, 200ug / L bFGF, 50ug / L SCF, 5×10 -8 mol / L dexamethasone.

[0091] Mesenchymal stem cell medium (configuration medium C): 450mLDMEM basal medium, 50mLFBS, 100ng / mLhEGF, 100ng / mL bFGF, 50ng / mL HGF, 20ng / mLPDGF, 25ng / mLTGF-β.

[0092] 2. Acquisition of urine cells

[0093] 1) Add 2mL penicillin / streptomycin double antibody to each collection cup.

[0094] 2) Collect the urine. If the follow-up operation is not performed immediately, store the urine in a 4°C refrigerator and complete the follow-up operation on the same day.

[0095] 3) One well of a six-well plate was prepared for each urine, and the well was coated with 0.1% gelatin for more than 20 minutes, and the liquid in the well was b...

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Abstract

The invention relates to the technical field of cell and gene engineering, in particular to a medium, a purpose thereof and a preparation method of mesenchymal stem cells. The invention provides the medium and a method of using the medium to induce urinary cells into the mesenchymal stem cells. The urinary cells are wide in source and are not limited by ethic, so that by adopting the method provided by the invention, the urinary cells are induced into the mesenchymal stem cells after forming IPS cells; and all induction can be finished within 12-15 days, the obtained cells subjected to detection accord with the characteristics of the mesenchymal stem cells and have a larger amount and higher 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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IPC IPC(8): C12N5/0775C12N5/10
CPCC12N5/0662C12N2500/32C12N2501/11C12N2501/115C12N2501/12C12N2501/125C12N2501/135C12N2501/15C12N2501/33C12N2501/39C12N2506/45
Inventor 黄燕飞车七石赵澎刘少辉
Owner GUANGZHOU RAINHOME PHARM&TECH CO LTD
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