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Method for preparing stem cell patch for treating severe skin diseases and injuries

A skin disease and stem cell technology, applied in medical science, bandages, absorbent pads, etc., can solve the problems of difficult to maintain cell polarity, limited survival time, dislocation of growth, etc., to prolong the survival time, repair broken skin, and repair scars Effect

Inactive Publication Date: 2014-12-24
奥思达干细胞有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the survival time of mesenchymal stem cells directly transplanted to the wound surface is relatively limited, and they are prone to apoptosis, growth dislocation and extremely difficult to maintain the polarity of cells after contact with the environment

Method used

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  • Method for preparing stem cell patch for treating severe skin diseases and injuries
  • Method for preparing stem cell patch for treating severe skin diseases and injuries
  • Method for preparing stem cell patch for treating severe skin diseases and injuries

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 The preparation of a simple polymer polyglycol (PCL)-collagen composite film, that is, a biofilm, the preparation method includes the following steps:

[0033] 1. Preparation of electrospinning solution: weigh 0.50g of PCL particles, dissolve in 4ml of chloroform; weigh another 0.025g of collagen, dissolve in 1ml of hexafluoroisopropanol (HFIP), and stir overnight with a magnetic stirrer (more than 12h). The two dissolved solutions were mixed and stirred with a magnetic stirring bar for more than 12 hours to obtain an electrospinning solution.

[0034] 2. Electrospinning process: put the electrospinning solution into the syringe, and the syringe is equipped with a blunt rotor with a diameter of 0.6mm. At a voltage of 10kv, the collection distance, that is, the distance between the nozzle and the receiving plate, is 15cm, and the liquid flow velocity is Electrospun under the condition of 0.75ml / h.

[0035] 3. Post-processing: After the electrospinning is comp...

Embodiment 2

[0037] Example 2 Culture and Screening of Mesenchymal Stem Cells

[0038] 1. Culture of mesenchymal stem cells

[0039] Transfer the culture medium containing mesenchymal stem cells into a centrifuge tube and centrifuge at 1000 rpm for 5 minutes; discard the supernatant, add DMEM culture medium containing 10% fetal bovine serum and 1% penicillin and streptomycin to the cells Mix well; transfer to a culture flask and place at 37°C, 5% CO 2 Incubator incubation. The medium was changed the next day, and the bottom of the bottle was covered in 4 to 7 days, and the cells were passaged when the degree of cell confluence reached about 80%. Take the cultured 3rd-5th passage cells for later use.

[0040] The mesenchymal stem cell suspension was inoculated in a 100mm culture dish. After 12 hours, the cells adhered to the wall. After the attachment, triangular and spindle-shaped cells could be seen growing on the wall. The medium was changed to remove unattached cells. After 4-6 days...

Embodiment 3

[0044] Example 3 Growth of mesenchymal stem cells on PCL-collagen composite membrane

[0045] 1. Fluorescein diacetate (FDA) staining of cells

[0046] Spread a PCL-collagen composite membrane with a cut diameter of 1.56cm in a 24-well culture plate, and seed the mesenchymal stem cell suspension evenly on the PCL-collagen composite membrane at a cell seeding density of 8000 / cm 2 , make it adsorb for 2 hours and then add the medium, the cell culture medium is DMEM medium (containing 10% fetal bovine serum + 1% penicillin and streptomycin). Dissolve fluorescein diacetate (FDA) in acetone at a concentration of 5 mg / ml, and dilute to 10 μg / ml with culture medium when used. Before staining, the cells grown on the membrane were washed twice with PBS buffer, then covered with FDA solution, placed in the incubator for half an hour, and observed under an inverted fluorescent microscope. Only living cells can be observed bright green. After the cells were inoculated into the pores of...

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Abstract

The invention discloses a stem cell patch for treating severe skin diseases and injuries and a preparation method thereof, and belongs to the field of regenerative medicine and tissue engineering. The patch is constructed by combination of an electrospinning biological membrane and a mesenchymal stem cell, so that the mesenchymal stem cell can be transplanted to a skin burst position to promote adhesion and proliferation of the mesenchymal stem cell, and the electrospinning biological membrane is capable of protecting cell growth to promote formation of microstructure and appearance of the mesenchymal stem cell and promoting healing of skin wounds, and a novel thought is provided to improvement of transplanting efficacy of the mesenchymal stem cell. According to the preparation method disclosed by the invention, materials are convenient to obtain, and the preparation is simple.

Description

technical field [0001] The invention belongs to the fields of regenerative medicine and tissue engineering, and in particular relates to a stem cell patch for treating severe skin diseases and injuries and a preparation method thereof. Background technique [0002] Various severe skin ulceration diseases and burns and scalds are common accidental injuries in life. Squamous cell carcinoma is a common malignant skin tumor, which can be ulcerated in the early stage, and often secondary to chronic ulcers or chronic sinus openings, or ulcers in scars that persist for a long time, resulting in local spread and distant metastasis. Effective clinical treatment should be carried out as soon as possible. Burning with boiling water, hot oil, and hot steam is a common occurrence, especially if a large area of ​​damage invades the dermis, which may cause prolonged healing; even if it heals, it will leave scar contractures and cause dysfunction; Can cause infection and even be life thre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L15/40A61L15/32A61L15/26A61L15/42
Inventor 周萱
Owner 奥思达干细胞有限公司
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