Application of Dickkopf-1 protein in preparation of medicament for treating X-linked hypophosphatemia rickets
A technology for hypophosphatemia and rickets, applied in the field of biomedicine, can solve problems such as no clinical treatment methods, and achieve the effect of treating X-linked hypophosphatemia rickets
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Embodiment 1
[0018] Mouse Osteoblast (MC3T3-E1) Cell Culture and Measurement of Extracellular Collagen Deposition
[0019] Experimental method: Mouse osteoblasts (MC3T3-E1) were obtained from Yuchun Biotechnology Co., Ltd. (Yuchun Biology, Shanghai), cultured in α-basic medium (α-MEM), supplemented with 10% fetal bovine serum and 1 % penicillin, streptomycin, cultured in 5% carbon dioxide, 95% oxygen, and 37°C, and replaced with fresh culture medium every 3-5 days to keep the culture medium in a subsaturated state. In preparation for the mineralization experiment, count the cells with a cell counter, then add 20,000 cells to each well of a 24-well culture plate and culture to 80% saturation. Then add 5mMβ-glycerophosphate and 100μg / ml ascorbic acid (called mineralization medium) to the medium, and change the medium every 3 days. In order to prove that the cells have the ability to secrete extracellular matrix mineralization, the cells cultured in the normal medium (normal medium) and the ...
Embodiment 2
[0022] Experimental method: Dkk1 protein was purchased from R&D Biosystems, USA. The cells cultured in Example 1 were divided into four groups: the first group was normal medium cultured cells without adding any reagents; the second group was mineralization medium control, added 0.2 μg / ml, 0.4 μg / ml and 0.8 Normal saline at the dose of μg / ml; the third group was the first day of the experiment, and 0.2 μg / ml, 0.4 μg / ml and 0.8 μg / ml of Dkk1 were added to the mineralization medium; the fourth group was the tenth day of the experiment Day, 0.2 μg / ml, 0.4 μg / ml and 0.8 μg / ml doses of Dkk1 were added to the mineralization medium. On the fourteenth day of the experiment, the medium for the experiment was collected, and the cultured cells were also collected in TRIzol reagent for future use. ELISA kits were used to detect the levels of mouse P1NP (procollagen I-type n-terminal propeptide, Elabscience, China) and mouse OCN (osteocalcin, Elabscience, China), and the experimental resu...
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