Preparation for promoting fibroblasts to secrete extracellular matrix components and preparation method thereof
A fibroblast, extracellular matrix technology, applied in the directions of medical preparations containing active ingredients, peptide/protein ingredients, preparations for skin care, etc., to inhibit or kill microorganisms, improve cell growth, and have a wide range of applications.
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Embodiment 1
[0019] Example 1. Preparation and RT-PCR detection of preparations for promoting fibroblasts to secrete extracellular matrix components
[0020] Take the egg white of fresh eggs (Bailaihang, Shouguang or Bailock) and let it stand overnight at 4°C, centrifuge at 4000r / min for 10min to get 2mg of the supernatant, dilute it with ultrapure water at a volume ratio of 1:10, and add 200mg / ml of egg white lysozyme and 20 mg / ml of sodium pyruvate were mixed to obtain the preparation for promoting fibroblasts to secrete extracellular matrix components provided by the present invention.
[0021] Take a Kunming white mouse with a gestational age of 14.5 days, remove the limbs and viscera, take the back skin, and cut it into pieces.
[0022] Soak 600U / ml collagenase and 400U / ml hyaluronidase overnight at 4°C, digest at 37°C for 1 hour, pass through a 200-mesh cell sieve to obtain mouse skin fibroblast suspension, add 10% fetal bovine serum to DMEM medium, and pass passage After purificat...
Embodiment 2
[0037] Example 2. Preparation of a preparation for promoting fibroblasts to secrete extracellular matrix components and the effect of the preparation on the content of hydroxyproline in mouse skin fibroblasts
[0038] Take the egg white of fresh eggs (both Bailaihang, Shouguang and Bailock) and let it stand overnight at 4°C, centrifuge at 4000r / min for 10min to get 4mg of the supernatant, dilute it with ultrapure water at a volume ratio of 1:10, and add 400mg / ml of egg white lysozyme and 5 mg / ml of sodium pyruvate were mixed to obtain the preparation provided by the invention for promoting the secretion of extracellular matrix components by fibroblasts.
[0039] Take a Kunming white mouse with a gestational age of 14.5 days, remove the limbs and viscera, take the back skin, and cut it into pieces.
[0040]Soak 600U / ml collagenase and 400U / ml hyaluronidase overnight at 4°C, digest at 37°C for 1 hour, pass through a 200-mesh cell sieve to obtain mouse skin fibroblast suspension...
Embodiment 3
[0044] Example 3. Preparation of a preparation for promoting the secretion of extracellular matrix components by fibroblasts and the effect of the preparation on the content of hydroxyproline in human skin fibroblasts
[0045] Take fresh eggs (Bailaihang, Shouguang or Bailock can all be used), take the egg white and let it stand overnight at 4°C, centrifuge at 4000r / min for 10min to get 0.75mg of the supernatant, dilute it with ultrapure water at a volume ratio of 1:10, and add 15mg Egg white lysozyme / ml and 0.2 mg / ml sodium pyruvate were mixed to obtain the preparation for promoting fibroblasts to secrete extracellular matrix components provided by the present invention.
[0046] Culture of human skin fibroblasts: DMEM medium, add 10% fetal bovine serum, add 5% of the preparation obtained by the above preparation method in the medium, as the preparation group; in the control group, the culture medium is DMEM medium, add 10% Fetal bovine serum, culture medium was changed every...
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