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Preparation method and application of novel modified egg white cell culture scaffold material

A cell culture and scaffold material technology, which is applied in the field of preparation of new modified egg white cell culture scaffold materials, and can solve problems such as poor stability

Active Publication Date: 2016-02-03
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the stability of genipin at high temperature is not very good, so it is recommended to dissolve genipin in water at about 37°C, and its solubility at this time is about 2%.

Method used

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  • Preparation method and application of novel modified egg white cell culture scaffold material
  • Preparation method and application of novel modified egg white cell culture scaffold material
  • Preparation method and application of novel modified egg white cell culture scaffold material

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preparation example Construction

[0033] A preparation method for a novel modified egg white cell culture support material, comprising the following steps:

[0034] (1) Add geninpin into the ultrapure water solvent and prepare 6 concentration gradients of 10%, 5%, 1%, 0.5%, 0.25%, and 0.125% by mass percentage respectively. For higher concentration of genipin (greater than 4%), the solution can be preheated to about 50° C. before adding genipin, stirring rapidly and then dissolving. After stirring, ultrasonically oscillate for 10-30min, put into 37℃ to fully dissolve, then take 0.2mL each and add to a 6-well plate with 2mL fresh egg white in each well, stir and mix well;

[0035] Other containers (such as beakers, petri dishes, etc.) can be selected during preparation, not limited to 6-well plates. After the preparation is completed, it can be added to the corresponding cell culture plate, bottle, dish, etc. The concentration of genipin is 1%, 0.5%, 0.25%, and 0.125% as the most suitable range, and 0 concent...

Embodiment 1

[0043] A preparation method for a novel modified egg white cell culture support material, comprising the following steps:

[0044] 1) Add 0.0125g of genipin to 9.9875mL of ultrapure water, prepare it to a concentration of 0.125%, stir, oscillate ultrasonically for 10-30min, put it in 37°C to fully dissolve, then add 0.2mL to each well and add 2mL to each well In a 6-well plate with fresh egg whites, stir well;

[0045] 2) Put the egg white added with the cross-linking agent genipin into a 37°C incubator for 24 hours to make it fully cross-linked;

[0046] 3) The modified egg white culture scaffold material obtained in step (2) was dripped into the PBS solution drop by drop with a pipette gun, and stood still for 12 hours, so that the excess genipin molecules therein were sucked out and removed, and repeated three times;

[0047] 4) After the reaction, the obtained modified egg white cell culture scaffold material was placed on a cell ultra-clean workbench, and irradiated with...

Embodiment 2

[0050] A preparation method for a novel modified egg white cell culture support material, comprising the following steps:

[0051] 1) Add 0.025g of genipin to 9.975mL of ultrapure water, prepare to a concentration of 0.25%, stir, oscillate ultrasonically for 10-30min, put it in 37°C to fully dissolve, then add 0.2mL to each well to contain 2mL In a 6-well plate with fresh egg whites, stir well;

[0052] 2) Put the egg white added with the cross-linking agent genipin into a 37°C incubator for 24 hours to make it fully cross-linked;

[0053] 3) The modified egg white culture scaffold material obtained in step (2) was dripped into the PBS solution drop by drop with a pipette gun, and stood still for 12 hours, so that the excess genipin molecules therein were sucked out and removed, and repeated three times;

[0054] 4) After the reaction, the obtained modified egg white cell culture scaffold material was placed on a cell ultra-clean workbench, and irradiated with ultraviolet lig...

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Abstract

The invention relates to a preparation method and application of a novel modified egg white cell culture scaffold material. The method includes: step one, adding genipin into sterile ultrapure water, stirring, oscillating ultrasonically and completely, thoroughly dissolving at 25-40 DEG C, adding 0.2ml of the genipin solution into a reaction vessel containing 2ml of fresh egg white, and well mixing; step two, putting the egg white with the genipin serving as a crosslinking agent into a thermostat to realize complete crosslinking; step three, using a pipette for dripping a modified egg white cell culture scaffold material obtained at the step two into a PBS solution drop by drop to guarantee immersion of materials without overflowing, standing for more than 12 hours to remove surplus genipin molecules, and repeating for several times; step four, putting a modified egg white cell culture scaffold material obtained after reaction on a cell clean bench, and performing ultraviolet radiation for sterilization to obtain the novel modified egg white cell culture scaffold material. The hardness gradient of the novel modified egg white cell culture scaffold material is easy to control, and safety and avoidance of toxic and side effects are realized.

Description

technical field [0001] The invention relates to a preparation technology of a novel modified egg white cell culture support material, in particular to a method for preparing a novel modified egg white cell culture support material with multiple hardness gradients by using a genipin crosslinking agent crosslinking method. Background technique [0002] Scaffold materials for tissue engineering are generally divided into natural materials and synthetic materials. Pure synthetic materials can easily induce inflammation, and toughness is difficult to guarantee. Natural materials such as collagen, chitosan, and hyaluronic acid are easy to degrade and have certain biological functions. According to different research purposes, natural materials can be chemically modified to change and optimize their material morphology and mechanical properties for tissue engineering needs. [0003] As a natural polymer, protein has good biodegradability and biocompatibility, and has wider applic...

Claims

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Application Information

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IPC IPC(8): C12N5/07C12N5/09
Inventor 张天柱郭振超杨新明房坤田吉来刘培党顾宁
Owner SOUTHEAST UNIV
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