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Adhesive bandage type artificial active tissue constructed by using culture solution without serum or bovine pituitary extracts and construction method of adhesive bandage type artificial active tissue

A bovine pituitary and culture medium technology, which is applied in the field of tissue engineering and medical wound repair, can solve the problems of unsatisfactory product timeliness, etc., and achieves the effects of being beneficial to skin regeneration, long shelf life, and easy to prepare.

Active Publication Date: 2015-10-14
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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  • Adhesive bandage type artificial active tissue constructed by using culture solution without serum or bovine pituitary extracts and construction method of adhesive bandage type artificial active tissue
  • Adhesive bandage type artificial active tissue constructed by using culture solution without serum or bovine pituitary extracts and construction method of adhesive bandage type artificial active tissue
  • Adhesive bandage type artificial active tissue constructed by using culture solution without serum or bovine pituitary extracts and construction method of adhesive bandage type artificial active tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1 cell culture fluid preparation

[0045] Cell culture medium A (artificial tissue fibroblast culture medium): use the basic culture medium DMEM (HyClone, USA), add penicillin and streptomycin dual-antibody 1~5000U / ml in the basic culture medium DMEM;

[0046] Cell culture medium B (artificial tissue epidermal cell culture medium): It is composed of the following substances by volume percentage: basic culture medium DMEM60%~85%; basic culture medium HAM's F-12 (ScienCell, USA)10%~40%; the following components 5% to 15%: penicillin and streptomycin double-antibody 1-5000U / ml; amphotericin B0.5-6000ng / 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 volume fractions of the three is 100%. The above culture medium DMEM and HAM's F-12 are all added with the corresponding volume of liquid according to the formula, penicillin, streptomycin double antibody, amphot...

Embodiment 2

[0047] Example 2 Separation and cultivation of dermal fibroblasts

[0048] 1) Materials:

[0049] 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, cells can be derived from embryonic, neonatal or older individuals, including adult donor tissue, embryonic progenitor cells, such as mesenchymal stem cells, can be used in the present invention and induced to differentiate, develop into the desired tissue. Fibroblasts are preferably male neonatal foreskin and autologous skin.

[0050] The epithelial source isolated by surgery was placed in sterile saline and sent to the cell culture room.

[0051] 2) digestion

[0052] Carefully remove subcutaneous fat tissue, fascia and capillaries with scissors, rinse twice with phosphate-buffered saline (1×PBS), cut into about 0.5 cm...

Embodiment 3

[0057] The separation of embodiment 3 epidermal cells

[0058] 1) Materials:

[0059] 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, cells can be derived from embryonic, neonatal or older individuals, including adult donor tissue, embryonic progenitor cells, such as mesenchymal stem cells, can be used in the present invention and induced to differentiate, develop into the desired tissue. Human epidermal cells can be selected from body skin or allogeneic skin.

[0060] The epithelial source isolated by surgery was placed in sterile saline and sent to the cell culture room.

[0061] 2) digestion

[0062] 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%...

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Abstract

The invention discloses an adhesive bandage type artificial active tissue constructed by using a culture solution without serum or bovine pituitary extracts and a construction method of the adhesive bandage type artificial active tissue. A cell culture solution without the serum or bovine pituitary extracts is adopted, and a biological film carrier is specially treated to ensure that a niche suitable for cell interaction is formed on a biological film, and promote human fibroblasts and epidermal cells to give play to self potentials, proliferate, disintegrate and excrete growth factors, so that an artificial tissue with active cells can be formed on the biological film in 4-7 days. The artificial tissue is high in cell activity and strong in division and migration ability, can home back to wound surfaces, and can promote tissue regeneration by serving as seed cells for curing burns and chronic ulcer. The adhesive bandage type artificial active tissue disclosed by the invention completely avoids immunological rejection of animal derived products including serum, pituitary extracts and collagen and the risk of infection of viruses transmitted to humans from animals, and a thin cell tissue taking the biological film as a scaffold and an infection barrier is formed, so that the adhesive bandage type artificial active tissue with biological safety is truly realized.

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