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Culture medium of sweat gland cells and preparation method thereof

A sweat gland cell and culture medium technology, applied in cell culture active agents, biochemical equipment and methods, tissue culture, etc., can solve the problems that affect the safety of clinical application and the low probability of sweat gland cell differentiation, achieve stable passage and increase proliferation Speed, the effect of increasing the induction

Active Publication Date: 2019-02-19
GUANGZHOU RAINHOME PHARM&TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The current special medium for sweat gland cells has a low differentiation probability, which is difficult to meet the actual needs, and contains serum, which will bring the risk of animal-derived pathogens and affect the safety of clinical application

Method used

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  • Culture medium of sweat gland cells and preparation method thereof
  • Culture medium of sweat gland cells and preparation method thereof
  • Culture medium of sweat gland cells and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1. Experimental group and control group settings:

[0033] 1. Culture medium of the experimental group:

[0034] Experimental group culture medium comprises basal culture fluid and supplement, and described basal culture fluid is SGDM culture fluid, and volume is 500mL; In terms of the addition amount in described basal culture fluid, described supplement comprises the following components:

[0035] ingredient name

working concentration

NGF

10ng / mL

TGF-β

10ng / mL

EGF

10ng / mL

heparan sulfate glycoprotein

10ng / mL

bone morphogenetic protein 4

10μg / L

[0036] Preparation method: Add 5 μg of NGF, 5 μg of TGF-β, 5 μg of EGF, 5 μg of heparan sulfate glycoprotein, and 45 μg of bone morphogenetic protein into 500 mL of basal culture medium according to the above ratio.

[0037] 2. Control medium

[0038] The control group used 500 mL of the existing SGDM culture medium, and added 10 mL of FBS to it.

[00...

Embodiment 2

[0050] In this embodiment, a method for inducing sweat gland cells by induced pluripotent stem cells is the same as step 2 in Example 1, the difference is that the culture medium of the experimental group includes basal culture fluid and additives, and the basal culture fluid is SGDM The culture fluid has a volume of 500 mL; based on the amount added in the basal culture fluid, the additive includes the following components:

[0051] ingredient name

[0052] The identification method of the cultured sweat gland cells is the same as Step 3 in Example 1, and the expression of K14, K18, CEA and K8 is similar to that observed under a fluorescent microscope figure 2 , count the cells emitting fluorescence, the results are as follows Figure 4 shown.

Embodiment 3

[0054]In this embodiment, a method for inducing sweat gland cells by induced pluripotent stem cells is the same as step 2 in Example 1, the difference is that the culture medium of the experimental group includes basal culture fluid and additives, and the basal culture fluid is SGDM The culture fluid has a volume of 500 mL; based on the amount added in the basal culture fluid, the additive includes the following components:

[0055]

[0056]

[0057] The identification method of the cultured sweat gland cells is the same as Step 3 in Example 1, and the expression of K14, K18, CEA and K8 is similar to that observed under a fluorescent microscope figure 2 , count the cells emitting fluorescence, the results are as follows Figure 5 shown.

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PUM

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Abstract

The invention relates to a culture medium for sweat gland cells and a preparation method of the culture medium. The culture medium comprises a basic culture solution and an additive, wherein the basic culture solution is an SGDM culture solution; the additive comprises the following components, by the addition amount in the basic culture solution, nerve growth factors, transforming growth factors-beta, epidermal growth factors, HEPG (heparin sulfate proteoglycan) and bone morphogenetic protein 4. The culture medium can increase the probability at which ips cells are induced into sweat gland cells, stable passage can be guaranteed, the cell proliferation speed is increased, and a source of seed cells is provided for skin tissue engineering. Additionally, no serum is used in the whole process, risks caused by animal origin pathogens are effectively avoided, and the clinical application safety is improved.

Description

technical field [0001] The invention relates to a cell culture medium, in particular to a sweat gland cell culture medium and a preparation method thereof. Background technique [0002] Skin tissue engineering is a research hotspot in recent years. However, related skin cells are all terminally differentiated cells, and it is difficult to perform large-scale culture and expansion in vitro. At the same time, the source of seed cells also hinders the development of skin tissue engineering. [0003] Due to the limited sources of autologous seed cells, rejection of allogeneic seed cells, and ethical issues with stem cells, urine cells are collected from urine and induced to become induced pluripotent stem cells (ips cells), ips cells Then induce keratinocytes, fibroblasts, sweat gland cells, dermal papilla cells, etc., to solve the problem of the source of seeds for skin tissue engineering, and obtain cells from urine, which is convenient and quick, without trauma, and has a wi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0633C12N2501/11C12N2501/13C12N2501/15C12N2501/155C12N2501/998C12N2506/03
Inventor 车七石
Owner GUANGZHOU RAINHOME PHARM&TECH CO LTD
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