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Culture medium and method for inducing fibroblast differentiation into sweat gland cells

A technology of fibroblasts and sweat gland cells, applied in the field of stem cell differentiation, can solve the problems of restricting the rapid development of research work and increasing the workload of research, and achieve the effect of avoiding ethical concerns, consistent differentiation stages, and good growth

Active Publication Date: 2020-09-15
GUANGZHOU RAINHOME PHARM&TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, in the traditional preparation method of sweat gland cells, sweat gland cells and stem cells need to be co-cultured, which not only increases the workload of research, but also limits the rapid development of research work due to the limited source of sweat gland cells

Method used

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  • Culture medium and method for inducing fibroblast differentiation into sweat gland cells
  • Culture medium and method for inducing fibroblast differentiation into sweat gland cells
  • Culture medium and method for inducing fibroblast differentiation into sweat gland cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Preparation of culture medium:

[0039] Medium A (fibroblast medium): the cell base medium is H-DMEM, added with FGF with a final concentration of 55ng / mL, EGF with a final concentration of 25ng / mL, and a final concentration of 5% (g / mL) Glutamine (glutamine), insulin with a final concentration of 45ng / mL, and HGF with a final concentration of 10ng / mL, a total of 500mL.

[0040] Medium B (sweat gland cell induction medium and maintenance medium): the cell base medium is DMEM, the final concentration of KGF is 30 μg / mL, the final concentration of HGF is 35 ng / mL, the final concentration of insulin is 5 ng / mL, and the final concentration is 5 ng / mL. EGF at a concentration of 10 μg / mL, NGF at a final concentration of 30 μg / mL, and a final concentration of 5×10 -7 mol / L of triiodothyronine, the final concentration is 2.5×10 -4 mol / L adenine, a total of 500mL.

[0041] 2. Isolation and culture of fibroblasts

[0042] (1) Take the foreskin discarded after circumcision ...

Embodiment 2

[0053] 1. Preparation of culture medium:

[0054] Medium A (fibroblast medium): the cell base medium is H-DMEM, added with FGF at a final concentration of 20ng / mL, EGF at a final concentration of 1ng / mL, and a final concentration of 1% (g / mL) Glutamine, insulin with a final concentration of 25ng / mL, and HGF with a final concentration of 2ng / mL, totaling 500mL.

[0055] Medium B (Sweat Gland Cell Induction Medium and Maintenance Medium): Cell basal medium is DMEM, supplemented with KGF at a final concentration of 12 μg / mL, HGF at a final concentration of 5 ng / mL, and insulin at a final concentration of 1 ng / mL , EGF with a final concentration of 1 μg / mL, NGF with a final concentration of 18 μg / mL, and a final concentration of 1×10 -7 mol / L of triiodothyronine, the final concentration is 2×10 -4 mol / L adenine, a total of 500mL.

[0056] 2. Isolation and culture of fibroblasts

[0057] (1) Take the discarded foreskin after circumcision of 3-6-year-old healthy children, and wa...

Embodiment 3

[0068] 1. Preparation of culture medium:

[0069] Medium A (fibroblast medium): the cell base medium is H-DMEM, added with FGF with a final concentration of 100ng / mL, EGF with a final concentration of 50ng / mL, and a final concentration of 10% (g / mL) Glutamine, insulin with a final concentration of 80ng / mL, and HGF with a final concentration of 15ng / mL, totaling 500mL.

[0070] Medium B (sweat gland cell induction medium and maintenance medium): DMEM as the cell base medium, KGF with a final concentration of 55 μg / mL, HGF with a final concentration of 70 ng / mL, insulin with a final concentration of 10 ng / mL, and EGF at a concentration of 20 μg / mL, NGF at a final concentration of 55 μg / mL, and a final concentration of 7.5×10 -7 mol / L triiodothyronine, the final concentration is 5×10 -4 mol / L adenine, a total of 500mL.

[0071] 2. Isolation and culture of fibroblasts

[0072] (1) Take the discarded foreskin after circumcision of 3-6-year-old healthy children, and wash it thre...

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Abstract

The invention provides a culture medium and a method for inducing fibroblasts to be differentiated into sweat gland cells. The culture medium is prepared by adding keratinocyte growth factor (KGF) with a final concentration of 12-55mug / mL, hepatocyte growth factor (HGF) with a final concentration of 5-70ng / mL, insulin with a final concentration of 1-10ng / mL, epidermal growth factor (EGF) with a final concentration of 1-20mug / mL, nerve growth factor (NGF) with a final concentration of 18-55mug / mL, triiodothyronine with a final concentration of 1-7.5*10<-7>mol / L, and adenine with a final concentration of 2-5*10<-4>mol / L into a basic cell culture medium. By adopting the culture medium with specific components and concentrations, the single type of fibroblasts can be directly induced to be differentiated into the sweat gland cells; the method is quick and efficient, and overcomes the defect that in the traditional technology, the sweat gland cells need to be co-cultured with cells such asstem cells, so that the yield of the sweat gland cells is limited.

Description

technical field [0001] The invention relates to the technical field of stem cell differentiation, in particular to a culture medium and a method for inducing fibroblast differentiation into sweat gland cells. Background technique [0002] The skin is the largest organ of the human body and plays an important role in defending against microbial invasion, ultraviolet radiation, preventing water loss, regulating body temperature, and is also one of the components of the immune system. The regeneration and repair of skin after trauma is mainly realized by the division and proliferation of adjacent and healthy cells. However, severe skin defects caused by large-area burns, extensive scar excision, traumatic skin defects, and skin ulcers, especially large-area burns, damage the entire layer of skin and its appendages, and it is difficult to achieve skin regeneration only by the wound itself. Thus adequate skin substitutes are required for repair. Although there are many tissue e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0633C12N2500/40C12N2501/11C12N2501/117C12N2501/12C12N2501/13C12N2501/33C12N2501/395C12N2506/1307
Inventor 黄燕飞车七石刘少辉
Owner GUANGZHOU RAINHOME PHARM&TECH CO LTD
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