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A wound-adhesive artificial active tissue constructed with serum-free and bovine pituitary extract culture fluid and its construction method

A technology of bovine pituitary gland and culture fluid, which is applied in the fields of tissue engineering and medical wound repair, which can solve the problems of unsatisfactory product timeliness, and achieve the effects of skin regeneration, avoiding immune rejection, and high cell activity

Active Publication Date: 2017-08-15
交发合锐生物技术(日照)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For autologous skin transplantation, it usually takes 15 to 20 days to separate epidermal cells from autologous skin until keratinization, which cannot meet the urgent needs of clinical skin grafting such as scalds and burns in terms of product timeliness.

Method used

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  • A wound-adhesive artificial active tissue constructed with serum-free and bovine pituitary extract culture fluid and its construction method
  • A wound-adhesive artificial active tissue constructed with serum-free and bovine pituitary extract culture fluid and its construction method
  • A wound-adhesive artificial active tissue constructed with serum-free and bovine pituitary extract culture fluid and its construction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1 cell culture fluid preparation

[0046] Cell culture medium A (artificial tissue fibroblast culture medium): use basic culture medium DMEM (HyClone, USA), add penicillin and streptomycin dual-antibody 1~5000U / ml in the basic culture medium DMEM;

[0047] Cell culture medium B (artificial tissue epidermal cell culture medium): composed of the following substances by volume percentage: basic culture medium DMEM 60% to 85%; basic culture medium HAM's F-12 (ScienCell, USA) 10% to 40%; Composition 5%~15%: penicillin, streptomycin double antibody 1~5000U / ml; amphotericin B 0.5~6000ng / ml; adenine 0.06~370ng / ml; insulin 0.02~530ug / ml; hydrocortisone 0.05 ~490ug / ml; triiodothyronine 0.03~280ng / ml; transferrin 0.07~460ug / ml; the sum of the three volume percentages is 100%, and the above culture medium DMEM and HAM's F-12 are added according to the formula Corresponding volume of liquid, penicillin, streptomycin double antibody, amphotericin B, adenine, insulin, hydroc...

Embodiment 2

[0048] Example 2 Separation and cultivation of dermal fibroblasts

[0049] 1) Materials:

[0050] Human fibroblasts can be derived from a variety of tissues including, but not limited to, male neonatal foreskin, dermis, tendon, lung, umbilical cord, cartilage, urethra, corneal stroma, oral mucosa, and intestine. Cell donors can vary in development and age, and cells can be derived from neonatal or older individuals, including adult donor tissues, such as mesenchymal stem cells, that can be used in the present invention and induced to differentiate into the desired tissue. Fibroblasts are preferably male neonatal foreskin and autologous skin.

[0051] The epithelial source isolated by surgery was placed in sterile saline and sent to the cell culture room.

[0052] 2) digestion

[0053] Carefully remove subcutaneous fat tissue, fascia and capillaries with scissors, rinse twice with phosphate-buffered saline (1×PBS), cut into about 0.5 cm strips, and soak in 0.05% trypsin at 4...

Embodiment 3

[0058] The separation of embodiment 3 epidermal cells

[0059] 1) Materials:

[0060] Human epidermal cells can be derived from a variety of tissues including, but not limited to, male neonatal foreskin, dermis, tendon, lung, umbilical cord, cartilage, urethra, corneal stroma, oral mucosa, and intestine. Cell donors can vary in development and age, and cells can be derived from neonatal or older individuals, including adult donor tissues, such as mesenchymal stem cells, that can be used in the present invention and induced to differentiate into the desired tissue. Human epidermal cells can be selected from body skin or allogeneic skin.

[0061] The epithelial source isolated by surgery was placed in sterile saline and sent to the cell culture room.

[0062] 2) digestion

[0063] Carefully cut off the subcutaneous fat tissue, fascia and capillaries with scissors, rinse twice with 1×PBS, cut into strips of about 0.5 cm, and soak in 0.05% trypsin at 4°C overnight.

[0064] 3)...

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Abstract

The invention discloses a wound-adhesive artificial active tissue constructed by using serum-free and bovine pituitary extract culture fluid and a construction method thereof. The present invention adopts the cell culture fluid without adding bovine serum and pituitary extract, and through special treatment of the biofilm carrier, a micro-environment (niche) suitable for cell interaction is formed on the biofilm to promote the development of human fibroblasts and epidermal cells. Potential, proliferation, division and secretion of growth factors, 4 to 7 days to form an artificial tissue with active cells on the biofilm. The artificial tissue cells have high activity, strong ability to divide and migrate, and can homing to the wound surface, as seed cells for healing burns and chronic ulcers, and promoting tissue regeneration. The invention completely avoids the immune rejection of animal-derived products such as serum, pituitary extract and collagen, and the risk of zoonotic virus infection, forms a thin layer of cell tissue with biofilm as a scaffold and infection barrier, and truly realizes a biosafe wound dressing artificial living tissue.

Description

technical field [0001] The invention relates to the technical fields of tissue engineering and medical wound repair, in particular to a serum-free and bovine pituitary extract cell culture fluid, a "band-aid" artificial living tissue formed on a medical biofilm using the culture fluid, and a construction method thereof. Background technique [0002] The skin is the largest organ of the human body and acts as a barrier to contact with the external environment. The skin is mainly composed of epidermis, dermis and subcutaneous tissue from the inside to the outside. There is a basement membrane between the epidermis and the dermis, and the subcutaneous tissue is rich in blood vessels and nerves. Skin defects can be caused by various reasons, such as burns, chronic diseases, trauma, etc. Although skin tissue has a strong regenerative ability, in the case of a large area of ​​full-thickness skin defect, the regenerative ability of the skin is significantly weakened, and a large n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A61L27/38A61L27/60
Inventor 孙涛
Owner 交发合锐生物技术(日照)有限公司
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