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Novel kit containing stem cell

A kit and stem cell technology, applied in the field of biomedicine, can solve problems to be developed, and achieve the effects of promoting proliferation, speeding up repair, and regulating cell proliferation and differentiation

Inactive Publication Date: 2018-05-22
FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, current treatments and drugs for skin damage are still to be developed

Method used

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  • Novel kit containing stem cell
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Obtaining and Identification of Exosomes

[0056] 1. Obtaining exosomes

[0057] (1) Provide mesenchymal stem cells

[0058] The morphology of umbilical cord mesenchymal stem cells figure 1 As shown, the mesenchymal stem cells in a good state of expansion and proliferation were taken, and the cells were digested into single cells with TrypLEExpress digestive enzyme; then, after washing with PBS containing 1% BSA, the preparation containing 5×10 6A single cell suspension of umbilical cord mesenchymal stem cells was labeled with CD90, CD73, CD105, CD34, CD45, CD14, CD19, and HLA-CR flow-type antibodies (BD Company, the United States), and the amount added to each tube was 5 μl at room temperature. Incubate with light for 20 minutes; wash 2 times with PBS containing 1% BSA, resuspend the cells to 400 μl, and perform flow cytometry, the results are as follows: figure 2 As shown, wherein (A) to (H) are HLA-CR, CD14, CD19, CD105, CD34, CD45, CD73, CD90 antibodie...

Embodiment 2

[0067] Embodiment 2 Preparation of the second reagent

[0068] (1) Soak fresh oxtail in 75% alcohol for 10 minutes, then rinse repeatedly with sterile saline;

[0069] (2) peel off the tail tendon, remove the sarcolemma and fascia, and place them in a sterile plate;

[0070] (3) Shred the tail tendon, immerse in 0.05mol / L acetic acid solution, and shake at 4°C for 48 hours;

[0071] (4) Move it into a pulverizer for pulverization, and then put it into a refrigerator at 4°C for secondary expansion;

[0072] (5) After 24 hours, filter with a 200-mesh sieve, collect the filtrate, and obtain a collagen solution;

[0073] (6) In a 6-well plate, add 5ml of collagen solution to each well to spread it in the well;

[0074] (7) Pre-freeze the orifice plate in a -30°C refrigerator for 6 hours, and then vacuum-dry it for 18-24 hours to obtain a collagen sponge;

[0075] (8) Place the collagen sponge in a 10 cm petri dish, add 30 mL of 0.25% glutaraldehyde solution to the petri dish f...

Embodiment 3

[0081] Example 3 in vitro induction of mesenchymal stem cells to differentiate into epidermal cells

[0082] 1. Differentiation of mesenchymal stem cells into epidermal cells in vitro

[0083] Cells were planted at a density of 6×10 on a 6-well plate pre-coated with Matrigel (BD Company, USA). 4 cells / cm 2 For umbilical cord mesenchymal stem cells, add 2ml of low-sugar DMEM medium containing 10% fetal bovine serum to each well, at 37°C, 5% CO 2 The cells were cultured in an incubator until the cells reached 50% confluency, and then 2 ml of epidermis induction conditioned medium was used for induction for 7 days, and the medium was changed every other day in half.

[0084] Epidermal induction conditioned medium formula: (DMEM:DF12=1:1 (purchased from Sigma), 20ng / ml epidermal growth factor (purchased from R&D), 15ng / ml basic fibroblast growth factor (purchased from R&D), 1% insulin-transferrin-selenium complex (purchased from Sigma), 0.1 μM dexamethasone (purchased from Sigm...

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Abstract

The invention discloses a kit. The kit includes: a first reagent including an exosome and a cytokine; and a second reagent including a biological scaffold and a stem cell. The kit can effectively promote the repairing of skin injuries and the healing of skin wounds.

Description

technical field [0001] The invention relates to the field of biomedicine. In particular, the present invention relates to a novel kit containing stem cells. Background technique [0002] As the largest organ of the human body, the skin has the functions of barrier, protection, temperature regulation and sensation. Skin defects caused by factors such as inflammation, ulcers, scalds, and burns can be life-threatening in severe cases. [0003] However, the current treatment methods and drugs for skin damage are still to be developed. Contents of the invention [0004] The present invention aims at solving one of the technical problems existing in the prior art at least to a certain extent. [0005] It should be noted that the present invention is based on the inventor's following findings: [0006] At present, autologous skin flaps or skin grafts are usually used to treat defect wounds in clinical practice, but there are problems such as new wound defects in the donor sit...

Claims

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Application Information

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IPC IPC(8): A61L27/24A61L27/38A61L27/54A61L27/60
CPCA61L27/60A61L27/24A61L27/3834A61L27/54A61L2300/414
Inventor 裴雪涛何丽娟张博文岳文王思涵房芳
Owner FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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