Humanized heterogenous cell epimatrix material and preparation method thereof

An exogenous matrix and humanized technology, applied in medical science, prosthesis, etc., can solve the problems of foreign body reaction, immune rejection, use restriction, etc., and achieve the effect of promoting healing, accelerating healing, and extending the shelf life

Active Publication Date: 2009-02-18
SHAANXI RUISHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the components of the heterogeneous extracellular matrix are mainly secreted and synthesized by animal cells, which are different from those of the human body. Therefore, it

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0018] 1) Acquisition of fibroblasts and epidermal cells: Aseptically obtain human skin tissue, remove the fat layer, cut into a size of 5mm×5mm, digest with neutral protease, separate the epidermis and dermis tissue; digest the dermis with 300U / ml collagenase Tissue, after digestion, the dermal tissue was blown away to collect the cell fluid, and the supernatant was discarded after centrifugation to obtain fibroblasts, which were inoculated into culture bottles to supplement the fibroblast culture medium, expanded and cultured to the required number; the isolated epidermis was treated with 0.25% After the trypsin digestion solution is digested, blow off and collect the cell fluid, centrifuge and discard the supernatant to obtain epidermal cells, inoculate into culture flasks to supplement the epidermal cell culture medium, and amplify and culture to the required number;

[0019] 2), the preparation of decellularized small intestinal submucosa: the porcine jejunum was rinsed wi...

example 2

[0025] 1) Adipose stem cell acquisition: Aseptically obtain human adipose tissue, cut into 1mm 3 Size, after digestion with 400U / ml collagenase solution, the adipose tissue was blown away to collect the cell solution, and the supernatant was discarded after centrifugation to obtain adipose stem cells; after adding the cell culture solution, the cells were suspended by blowing, inoculated into the culture bottle, and expanded to the desired size quantity;

[0026] 2) Preparation of acellular pigskin matrix: Cut the pig skin with the fat layer removed into 5cm×10cm thin slices, rinse it with distilled water, put it in an aqueous solution containing 0.2% peracetic acid and 3% ethanol for 1.5 hours and sterilize it ;After rinsing with phosphate buffer, freeze at -80°C for 40 minutes to make the temperature of the inside and outside of the pigskin reach the same level, take it out and thaw it naturally, repeat the freeze-thaw process 3 times, so that the cells are completely broken...

example 3

[0032] 1), the acquisition of fibroblasts: same as Example 1;

[0033] 2), the preparation of decellularized small intestinal submucosa: same as Example 1;

[0034] 3), preparation of special culture medium, each component by volume percentage is: commercial minimum necessary culture medium DMEM 67.5%, commercial culture medium F12 22.5%, fetal calf serum 10%, insulin 20 μ g / ml, basic fibroblast growth Factor 10ng / ml, hydrocortisone 200ng / ml, adenine 0.25mM, transferrin 10μg / ml, vitamin C 50μg / ml.

[0035] 4), preparation of humanized heterogeneous extracellular matrix material: suspend fibroblasts in cell culture medium, press 10 6 piece / cm 2 The density was dropped on the surface of the decellularized small intestinal submucosa in 5% CO 2 Stand still at 37°C for 30 minutes in the environment, add special culture medium to culture for 3 days, discard the culture medium, put the side of the decellularized small intestinal submucosa that is not planted with cells upward, and...

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Abstract

The invention relates to a humanized heterogenous extracellular matrix material, which is a dried cellular foam material made by compounding an extracellular matrix and a cell growth factor which are synthesized and excreted by human body cells on a heterogenous extracellular matrix. Cells and natural antigen components are removed from the prepared humanized heterogenous extracellular matrix material, and when the humanized heterogenous extracellular matrix material is applied to the surface of wound, the humanized heterogenous extracellular matrix material can directly take part in the repair of the surface of wound, and the cell growth factor contained in the humanized heterogenous extracellular matrix material can guide the ingrowth of the cells around the wound and creation of blood vessel, and induce the differentiation of stem cells to skin cells, thereby obviously promoting the healing of the surface of wound; besides, the prepared product is a dried product, which greatly prolongs storage life, and not only has part of the characteristics of human body tissues and good mechanical property of the heterogenous extracellular matrix, but also has higher biocompatibility to human body and obviously reduces the immune exclusive reaction; at the same time, the prepared product can cover the surface of wound, fill the defection of soft tissues, promote the growth and proliferation of the cells around the wound, repair the defection of the soft tissue organs, and promote the wound healing.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering biomaterials, in particular to a humanized heterogeneous extracellular matrix material and a preparation method thereof. Background technique [0002] The behavior of cells in organs and tissues not only depends on the genetic sequence inside the cells, but also is largely influenced by external environmental factors, including the interaction between cells and the extracellular matrix. The extracellular matrix not only provides support and protection for cell growth, but more importantly, the interaction between cells and extracellular matrix can regulate the morphogenetic process of cells and affect cell survival, migration, proliferation and functional metabolism. Therefore, it is very important and urgent to prepare extracellular matrix materials that are conducive to seed cell adhesion, proliferation and differentiation in tissue engineering research. [0003] Extracellular matrix...

Claims

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Application Information

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IPC IPC(8): A61L27/40A61L27/38A61L27/26A61L27/56
Inventor 王爱军
Owner SHAANXI RUISHENG BIOTECH
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