Human adipose-derived stem cell secreted polypeptide ADSCP2 and application thereof
A technology of human adipose stem cells and compositions, applied in the application, animal cells, animal/human peptides, etc., to achieve the effect of reducing the amount of myofibroblasts and reducing the content of collagen
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Embodiment 1
[0021] Entrust Shanghai Keyept Biotechnology Co., Ltd. to synthesize the following sequence shown in SEQ ID NO.1 by solid-phase method:
[0022] Asp Glu Asn Arg Glu Lys Val Asn Asp Gln Ala Lys Leu
[0023] This is a polypeptide composed of thirteen amino acids. The main biological parameters of the aforementioned polypeptide sequence obtained through online tools are as follows:
[0024] The isoelectric point (PI) is 4.78, and the molecular mass (Mw) is 1558.67Da.
[0025] The above sequence is a natural sequence derived from the culture supernatant of human adipose stem cells. In addition to obtaining by chemical solid-phase synthesis, it can also be obtained by enzymatic digestion of long-chain polypeptides, and can also be cloned and expressed by host microorganisms or cells. The DNA fragment carrying the nucleotide sequence encoding one of the polypeptides is prepared by existing recombinant DNA technology. The nucleotide sequence encoding the above polypeptide is: gatga...
Embodiment 2
[0028] 1. Test materials
[0029] The RNA reverse transcription kit and RNA detection kit were purchased from Nanjing Novozyme Company, and the fibroblast culture medium was purchased from Sciencell Company of the United States.
[0030] 2. Synthesis and dilution of ADSCP2 polypeptide
[0031] The polypeptide ADSCP2 obtained by solid-phase synthesis in Example 1 of this experiment is taken as an example. Take 10mg of polypeptide, dilute it with sterile double distilled water to 50mM-40°C and store it for later use.
[0032] 3. Real-time quantitative PCR (RT-PCR) detection of the effect of polypeptide ADSCP2 on the mRNA expression of collagen genes COL1A1, COL1A2, COL3A1 in human scar dermal fibroblasts and the main marker of myofibroblasts ACTA2
[0033] At 37°C, 5% CO 2 In the incubator, cultivate human scar dermal fibroblasts, press 10 5 Seed in 6-well plates at a density of 1 / mL, and when the cell confluency reaches about 60%, add polypeptides (final concentrations of 1...
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