Culture medium for sweat gland cells and preparation method of culture medium
A sweat gland cell and culture medium technology, which is applied to cell culture active agents, biochemical equipment and methods, animal cells, etc., can solve the problems of low sweat gland cell differentiation probability and affect the safety of clinical application, and achieve increased proliferation speed and stable Passage, improve the effect of induction
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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.
[0039] 2. Induction of iP...
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 working concentration NGF 5ng / mL TGF-β 5ng / mL EGF 5ng / mL heparan sulfate glycoprotein 5ng / mL bone morphogenetic protein 4 5μg / L
[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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