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Preparation method for temperature sensing cornea epithelial cell culture material and application

A corneal epithelial cell, sensitive technology, applied in the direction of animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of high cost of temperature-sensitive cell culture plates, to avoid cell function damage, broad application prospects, growth effect in good condition

Inactive Publication Date: 2010-05-05
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this technology has reached the international advanced level, the graft rejection and complications caused by the degradation of scaffold materials in vivo will limit its further development.
[0007] Nishida et al. are the pioneers of foreign carrier-free cell culture technology, but the cost of preparing temperature-sensitive cell culture plates by their research group using ion grafting is high

Method used

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  • Preparation method for temperature sensing cornea epithelial cell culture material and application
  • Preparation method for temperature sensing cornea epithelial cell culture material and application
  • Preparation method for temperature sensing cornea epithelial cell culture material and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The structural characterization of the azidoaniline derivative of embodiment 1N-polyisopropylacrylamide / acrylic acid

[0040] The reaction for preparing N-isopropylacrylamide / acrylic acid is a free radical polymerization reaction, once initiated, it goes through more thoroughly, and the residual amount of monomer is very small. In addition, during the experiment, the reaction ratio of the monomers was strictly controlled, and refined by methanol dilution and ether precipitation to remove unreacted monomers, initiators and chain transfer agents, so the influence of reactant residues on the test results can be basically eliminated . Use Japan Shimadzu LC-6A high performance liquid chromatograph to detect reactant (N-isopropylacrylamide monomer, acrylic acid, 3-mercaptopropionic acid, azobisisobutyronitrile) and product (N-isopropylacrylamide acrylamide / acrylic acid). High-performance liquid chromatography analysis conditions: at room temperature, use a 7125 manual injec...

Embodiment 2

[0044] Temperature-sensitive properties and turbidity of the azidoaniline derivative material of embodiment 2N-isopropylacrylamide / acrylic acid

[0045] image 3 It reflects the phase transition process of N-isopropylacrylamide in the range of 0-60℃. The aqueous solution of N-isopropylacrylamide is basically transparent below 30°C, and becomes turbid as the temperature rises, that is, phase separation occurs. As the temperature further increases, the degree of turbidity gradually increases until it becomes turbid around 40°C reached stability. When the temperature drops to 30°C again, the cloudy solution can become transparent again, it can be seen that its L°CST is in the range of 30-35°C. Since the main chain of the N-isopropylacrylamide molecule is connected with a carboxyl group, it has strong hydrophilicity, so that the solution is only turbid after the phase separation of the N-isopropylacrylamide side chain, and no precipitation occurs. The result of chain phase sepa...

Embodiment 3

[0050] Example 3 Thermosensitivity test of culture plates immobilized with N-isopropylacrylamide / acrylic acid azidoaniline derivatives

[0051] When preparing temperature-sensitive polystyrene culture plates, add the N-isopropylacrylamide / acrylic acid azidoaniline derivative solution at 20 μl / well and dry at 10°C. During the whole process of the ultraviolet photochemical grafting reaction, it is necessary to ensure that the experimental temperature is always lower than the LCST (21.5° C.) of the azidoaniline derivative of N-isopropylacrylamide / acrylic acid. The low concentration of sample addition is because the monomer concentration is too high, which will lead to increased homopolymerization and chain termination reactions, which will reduce the grafting rate. The reaction after drying is due to the strong absorption of ultraviolet light by the monomer containing azo chromophore, and the grafted film cannot be obtained by the usual liquid phase grafting method. Keep the rea...

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Abstract

The invention discloses a preparation method for temperature sensing cornea epithelial cell culture material and application. The method is as follows: combining nitrine-aniline derivative of N- isopropyl acrylamide / acrylic acid, and then fixing the nitrine aniline derivative of N-isopropyl acrylamide / acrylic acid on a polystyrene board used for tissue culture by a UV-light chemical graft method, thus obtaining the culture material. The invention also discloses the application of the temperature sensing cornea epithelial cell culture material in culturing human corneal epithelial cells. The temperature sensing cornea epithelial cell culture material prepared by the preparation method has good plasticity and biocompatibility, can support the good growth of the cornea epithelial cells, does not cause inflammatory reaction or cell functional damage, and has good clinical application prospect.

Description

technical field [0001] The invention relates to the field of artificial cornea preparation, in particular to a preparation method and application of a temperature-sensitive corneal epithelial cell culture material. Background technique [0002] Corneal disease is the second leading cause of blindness after cataracts. Penetrating keratoplasty is the main treatment method, but it has a poor prognosis for severe dry eye, Stevens-Johnson syndrome, severe chemical burns, etc., and faces problems such as graft rejection and insufficient donors. The application of artificial corneas and research on tissue engineering corneas have brought hope to patients with corneal diseases, but corneal construction materials are still important factors that limit the development of these two corneas (Shao Chunyi, Fu Yao, Fan Xianqun. Tissue engineered corneas and artificial corneal materials Research progress [J]. International Survey of Ophthalmology, 2007, 31(1): 29-33). [0003] Artificial ...

Claims

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Application Information

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IPC IPC(8): C12N5/071C08F220/54C08F220/06C08F8/32C08G81/02
Inventor 关燕清刘俊明
Owner SOUTH CHINA NORMAL UNIVERSITY
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