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Application of Wnt signal pathway activator to preparation of products for improving abnormal osteogenic differentiation ability of hypophospatasia stem cells

A signaling pathway and osteogenic differentiation technology, which is applied in the field of preparation of products that improve the abnormal osteogenic differentiation ability of stem cells in hypoalkaline phosphatase disease, to achieve the effect of improving osteogenic differentiation ability

Pending Publication Date: 2019-11-26
XIAN TISSUE ENG & REGENERATIVE MEDICINE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Existing treatment options can only slow down the onset time of HPP, but cannot effectively reverse the fact of bone loss, so they cannot fundamentally solve the primary problem of defects in bone formation and bone regeneration

Method used

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  • Application of Wnt signal pathway activator to preparation of products for improving abnormal osteogenic differentiation ability of hypophospatasia stem cells
  • Application of Wnt signal pathway activator to preparation of products for improving abnormal osteogenic differentiation ability of hypophospatasia stem cells
  • Application of Wnt signal pathway activator to preparation of products for improving abnormal osteogenic differentiation ability of hypophospatasia stem cells

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Experimental program
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Effect test

Embodiment 1

[0093] 1. Acquisition and detection of mesenchymal stem cells with abnormal differentiation ability in hypoalkaline phosphatase disease

[0094] 1. Acquisition of mesenchymal stem cells with abnormal differentiation ability in hypoalkaline phosphatase disease

[0095] Buffy coat cells were isolated from clinically discarded bone marrow tissues using Percoll separation medium. The steps are as follows: mix the diluted Percoll separation solution with the bone marrow sample, centrifuge at 2000rpm for 20min; after centrifugation, the liquid appears stratified, use a gun to suck the buffy coat cells into a new centrifuge tube, add PBS buffer to wash, and centrifuge at 1000rpm for 10min Discard the supernatant after centrifugation and wash the cells again with PBS buffer, centrifuge at 8000rpm for 5min; suspend the cells with cell culture medium after centrifugation, inoculate into α-MEM medium containing 5-10% fetal bovine serum for adherent culture , the cultivation environment ...

Embodiment 2

[0116] 1. Acquisition and detection of mesenchymal stem cells with abnormal differentiation ability in hypoalkaline phosphatase disease

[0117] 1. Acquisition of mesenchymal stem cells with abnormal differentiation ability in hypoalkaline phosphatase disease

[0118] Collect clinically discarded patients’ teeth, remove the connective tissue on the surface of the tooth in an ultra-clean workbench, rinse several times with PBS containing double antibodies, smash the tooth with a hammer, and separate the pulp tissue to an appropriate degree. Culture medium, blow gently with a gun to disperse the pulp tissue into small cell clumps or uniform single cell suspension, in α-MEM medium (gibco, 11900-024) containing 5-10% fetal bovine serum Adhesive culture, each passage time is 2-3 days, to obtain 1-5 passages of dental pulp stem cells with abnormal differentiation ability. The cultivation environment is: CO 2 Concentration is 5%, O 2 The concentration was 20%, and the temperature ...

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Abstract

The invention provides application of a Wnt signal pathway activator to preparation of products for improving the abnormal osteogenic differentiation ability of hypophospatasia stem cells. Cell experiments reveal that the osteogenic differentiation ability of the hypophosphatase dental pulp stem cells is lowered, a key regulatory factor of an intracellular classical Wnt signal pathway is inhibited, the activator LiCl of the classical Wnt signal pathway is used for acting on the dental pulp stem cells with the abnormal osteogenic differentiation ability, and differentiation characteristics of the dental pulp stem cells with the abnormal osteogenic differentiation ability are improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of a Wnt signaling pathway activator in the preparation of products for improving the abnormal osteogenic differentiation ability of stem cells in hypoalkaline phosphatase disease. Background technique [0002] Hypophosphatasia (HPP) is caused by a mutation in the liver / bone / kidney alkaline phosphatase gene (ALPL) that causes a defect in the activity of the encoded tissue-nonspecific alkaline phosphatase (TNAP) protein A genetic disorder that causes a decrease in the bone mass of the bones and teeth. The incidence of hypoalkaline phosphatase is high, ranging from 4 / 1000 in mild to moderate cases, and 1 / 100,000 in severe cases. In severe cases, bone deformities, hypercalcemia, severe epilepsy, and respiratory system complications may occur, resulting in perinatal death , a serious threat to human health and quality of life. At present, the clinical treatment of hypoa...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0663C12N2500/12
Inventor 张立强刘文佳
Owner XIAN TISSUE ENG & REGENERATIVE MEDICINE RES INST
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