CN101524063A - Cornea mid-term preservation liquid - Google Patents
Cornea mid-term preservation liquid Download PDFInfo
- Publication number
- CN101524063A CN101524063A CN200910134005A CN200910134005A CN101524063A CN 101524063 A CN101524063 A CN 101524063A CN 200910134005 A CN200910134005 A CN 200910134005A CN 200910134005 A CN200910134005 A CN 200910134005A CN 101524063 A CN101524063 A CN 101524063A
- Authority
- CN
- China
- Prior art keywords
- cornea
- liquid
- mid
- term
- preservation liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to a cornea mid-term preservation liquid. 1000ml of preservation liquid contains 0.5-1.5g of DMEM culture medium, 1-10g of hydroxypropyl methylcellulose, 0.1-10ml of 2-mercaptoethanol, 10-20g of dextran (with the molecular weight of 40000), 1-3ml of nonessential amino acid, 10-32mmol of Hepes buffer solution and 0.01-0.05mmol of dexamethasone, 50-70mg of gentamicin. The pH value of the preservation liquid is 7.2-8.0, and the permeation pressure is 350-400mmol/LH2O. The cornea mid-term preservation liquid not only has good cornea storage quality, but also has the advantage of good cost performance and is the cornea mid-term preservation liquid which is widely applicable to the eye bank at home and abroad.
Description
Technical field
The present invention relates to a kind of cornea and preserve liquid mid-term.
Background technology
The active store method of existing cornea according to the difference of holding time, can be divided into short-term preservation, preservation in mid-term and long preservation.It is after the mattress processing is killed in the whole eyeball sterilization, to preserve under the 4 ℃ of conditions in wet room that short-term is preserved, and the advantage of the method is that method is easy, and is effective and feasible, is particularly suitable for basic hospital and carries out corneal transplantation.Shortcoming is that the holding time is shorter, generally all in 48 hours effectively, sometimes because of can not in time finding the patient, material is out of date and abandon.Long preservation is through freezing preservation in-196 ℃ liquid nitrogen after the specially treated with corneal film.The great advantage of freezing preservation is to provide high-quality active cornea at any time for clinical, but this method needs special installation, technical difficulty height.Preserve the general corneal film tissue that adopts mid-term and preserve 4 ℃ of preservations of liquid, this method not only can provide the cornea of better quality, and corneal graft is improved aspect arrangement of time.At present, it is K-SOK that the most frequently used several cornea is preserved liquid mid-term, CSM, Dexsol and Optisol.It is similar that these preserve the liquid basic function, but composition and the concentration difference formed.They exist a common problem to be: they are imported product all, and it costs an arm and a leg.In recent years; for being provided to society, inexpensive cornea protects liquid mid-term; Chinese scholar has been done many researchs in this respect, has all solved this problem preferably as Chinese patent literature CN1162389, CN1262866 and the disclosed corneal storage medium of CN1631138.
Summary of the invention
The objective of the invention is to overcome the problems referred to above, provide a kind of new inexpensive cornea to protect liquid mid-term to society.
Technical scheme of the present invention is: provide a kind of cornea to preserve liquid mid-term, it is to contain in the preservation liquid with 1000ml: DMEM medium 0.5-1.5g, hydroxypropyl methylcellulose 1-10g, 2 mercapto ethanol 0.1-10ml, dextran (molecular weight 40000) 10-20g, dispensable amino acid 1-3ml, Hepes buffer solution 10-32mmol, dexamethasone 0.01-0.05mmol, gentamicin 50-70mg; This pH value of preserving liquid is 7.2-8.0, and osmotic pressure is 350-400mmol/LH
2O.
Described gentamicin 50-70mg can replace with the 50-70mg streptomycin.
Described gentamicin 50-70mg can replace with 20-40mg streptomycin and gentamicin 20-40mg.
The present invention is by the DMEM medium, hydroxypropyl methylcellulose, and 2 mercapto ethanol, dextran, composition adding distil waters such as dispensable amino acid are formulated.It is different from other cornea and preserves liquid mid-term, wherein hydroxypropyl methylcellulose can protect cornea not to be subjected to the injury of external force, and hydroxypropyl methylcellulose itself is a kind of inert substance, can reduce cellular metabolism speed, pair cell nonhazardous effect can improve colloid osmotic pressure; The 2 mercapto ethanol cell growth plays a very important role, it is equivalent to hyclone, the proliferation function that direct irritation cell is arranged, the active part of 2 mercapto ethanol can make the glutathione of preserving oxidation in the liquid become reduced glutathione, thus inducing cell propagation.The present invention is owing to adopted the hydroxypropyl methylcellulose and the 2 mercapto ethanol of less expensive, make the present invention not only have good preservation of cornea quality, the advantage that also has good cost performance simultaneously is that a kind of cornea that is widely used in that domestic and international eye bank uses is preserved liquid mid-term.
Description of drawings
Fig. 1 is human corneal endothelial cells trypan blue-alizarin red dyeing * 100 that cornea of the present invention is preserved liquid preservation 5d mid-term.
Fig. 2 is human corneal endothelial cells trypan blue-alizarin red dyeing * 100 that Optisol-GS liquid is preserved 5d.
Fig. 3 is human corneal endothelial cells trypan blue-alizarin red dyeing * 100 that cornea of the present invention is preserved liquid preservation 8d mid-term.
Fig. 4 is human corneal endothelial cells trypan blue-alizarin red dyeing * 100 that Optisol-GS liquid is preserved 8d.
Fig. 5 is human corneal endothelial cells trypan blue-alizarin red dyeing * 100 that cornea of the present invention is preserved liquid preservation 14d mid-term.
Fig. 6 is human corneal endothelial cells trypan blue-alizarin red dyeing * 100 that Optisol-GS liquid is preserved 14d.
Fig. 7 is people's cornea tissue section that cornea of the present invention is preserved liquid preservation 8d mid-term.
Fig. 8 is people's cornea tissue section that cornea of the present invention is preserved liquid preservation 14d mid-term.
Fig. 9-Figure 10 is human corneal endothelial cells central authorities and neighboring scan electron microscopic observation * 1000 that cornea of the present invention is preserved liquid preservation 10d mid-term.
Figure 11-Figure 12 is human corneal endothelial cells central authorities and neighboring scan electron microscopic observation * 1000 that Optisol-GS liquid is preserved 10d.
Figure 13 is human corneal endothelial cells scanning electron microscopic observation * 1000 that cornea of the present invention is preserved liquid preservation 14d mid-term.
Figure 14 is human corneal endothelial cells scanning electron microscopic observation * 1000 that Optisol-GS liquid is preserved 14d.
Figure 15 is human corneal endothelial cells transmission electron microscope observing * 5200 that cornea of the present invention is preserved liquid preservation 14d mid-term.
Figure 16 is human corneal endothelial cells transmission electron microscope observing * 5200 that Optisol-GS liquid is preserved 14d.
Embodiment
The DMEM medium (liquid) that is adopted among the present invention is produced by U.S. GIBCO company, it contains 1000mg/L glucose, the 584mg/L-glutamine, the 110mg/L pyruvic acid, the 3700mg/L sodium bicarbonate, the pH value is 7.2-7.4, and it is a kind of medium that many mammalian cells are cultivated that is very suitable for.Obviously, above-mentioned DMEM medium also can after the weight of calculating its component, directly replace the DMEM medium of liquid with the DMEM culture medium dry powder with identical or close component with dry powder DMEM medium.
(Hydroxypropyi methyleeilulose HPMC) is produced by U.S. DOW company hydroxypropyl methylcellulose among the present invention.
2 mercapto ethanol among the present invention, dextran and HEPES buffer solution are produced by U.S. Sigma company.
Dispensable amino acid among the present invention is the dispensable amino acid solution of being produced by U.S. HyClone company (10mM, 100 times), and it contains seven kinds of dispensable amino acid, every kind of each 10mM (dissolving in the MEM medium).2-8 ℃ of preservation.
Gentamicin, streptomycin, dexamethasone, hydrocortisone are homemade.
Table 1 is preserved the prescription of liquid mid-term for cornea of the present invention:
Respectively each cornea in the table 1 is preserved liquid and Optisol-GS liquid mid-term, respectively with 8 people's corneas, respectively with preserving test in 5 days, 8 days and 14 days, carry out penetrating keratoplasty at Shenzhen's ophthalmologic hospital, found that working as with the Optisol-GS liquid phase with interior every technology indication in 8 days preserving among the 1-12 embodiment of the present invention, Optisol-GS liquid that fully can substituting import one.When preserving the 14th day, the data of the 4th, 5,6 and No. 12 embodiment wherein also slightly are better than Optisol-GS liquid.
Through test, gentamicin in the table 1 or streptomycin 50-70mg are replaced with 20-40mg streptomycin and gentamicin 20-40mg, can play same antibacterial effect.
The cut into slices result of comparison of people's cornea that the 6th bugle film in the table 1 is preserved that liquid preserves mid-term and people's cornea of Optisol-GS liquid preservation is as follows.
1, preserves the endothelial cell activity rate: see Table 2.Learn by statistics and handle, preserve at two kinds and preserve different time endothelial cell activity rate in the liquid relatively, difference does not have significance (P＞0.05) (seeing Fig. 1～6).
Preservation different time human corneal endothelial cells activity rate comparison in two kinds of preservations of table 2 liquid (x ± s, %)
2, corneal thickness measurement result:
See Table 3.The average corneal central thickness of sheet of planting of preserving preservation 14d in the liquid at cornea of the present invention mid-term is 649.2 μ m, and the average corneal central thickness of sheet of planting of preserving 14d in Optisol-GS liquid is 655.1 μ m.Two groups of comparing differences do not have significance (p＞0.05).
Preservation different time people corneal thickness variation in two kinds of preservations of table 3 liquid (μ m, x ± s)
3, histological observation:
Preserve that 5,8,12,14 days corneal film institutional frameworks are clear (preserved inter-lamination of cornea clear in structure, complete, the no oedema of endodermis 8 days.Preserve and rose in 12 days, visible corneal stroma light-water is swollen, but epithelium, endodermis remain intact (seeing Fig. 7～8).
4, scanner uni transmission electron microscope observing
ESEM is checked: 10 days endothelial cell hexagon mosaic texture of two groups of preservation liquid preservations is normal, and cell connects comparatively tight, and the Y type connects non-cracking, and cell space is bigger, and parts of fine karyon projection is obvious; The endothelial cell of preserving 14 days connects still tightr, and indivedual small ceasmas appear being dispersed in cell membrane.Transmission electron microscope observing: 14 days endothelial cell film of two groups of preservation liquid preservations is still complete, and the part mitochondrial swelling has cavity (seeing Fig. 9-16) in the cell.
5, microorganism is cultivated and observes
Preserve liquid in the preservation process for two groups, the liquid clarification, the censorship sample, the bacterium inspection is all negative.
Claims (3)
1, a kind of cornea is preserved liquid mid-term, it is characterized in that: it is to contain in the preservation liquid of 1000ml: DMEM medium 0.5-1.5g, hydroxypropyl methylcellulose 1-10g, 2 mercapto ethanol 0.1-10ml, dextran (molecular weight 40000) 10-20g, dispensable amino acid 1-3ml, Hepes buffer solution 10-32mmol, dexamethasone 0.01-0.05mmol, gentamicin 50-70mg; This pH value of preserving liquid is 7.2-8.0, and osmotic pressure is 350-400mmol/LH
2O.
2, cornea according to claim 1 is preserved liquid mid-term, it is characterized in that: described gentamicin 50-70mg can replace with the 50-70mg streptomycin.
3, cornea according to claim 1 is preserved liquid mid-term, it is characterized in that: described gentamicin 50-70mg can replace with 20-40mg streptomycin and gentamicin 20-40mg.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101340052A CN101524063B (en) | 2009-03-23 | 2009-03-23 | Cornea mid-term preservation liquid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101340052A CN101524063B (en) | 2009-03-23 | 2009-03-23 | Cornea mid-term preservation liquid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101524063A true CN101524063A (en) | 2009-09-09 |
| CN101524063B CN101524063B (en) | 2012-06-13 |
Family
ID=41092271
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009101340052A Active CN101524063B (en) | 2009-03-23 | 2009-03-23 | Cornea mid-term preservation liquid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101524063B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102132697A (en) * | 2010-01-21 | 2011-07-27 | 周海华 | Amniotic membrane long-term preserving fluid and preparation method thereof |
| CN105284788A (en) * | 2015-11-20 | 2016-02-03 | 厦门大学 | Cornea metaphase preservation liquid and preparation method thereof |
| CN106900695A (en) * | 2017-04-01 | 2017-06-30 | 北京焕生汇生物技术研究院 | A kind of mankind's inductive pluripotent stem cells are special to freeze reagent and cryopreservation methods |
| WO2020019516A1 (en) * | 2018-07-26 | 2020-01-30 | 姚晓明 | Cornea medium-term preserving fluid |
| CN111802378A (en) * | 2020-07-24 | 2020-10-23 | 镇江雷音再生医学科技有限公司 | SMILE (Small Scale Integrated Circuit) -derived protective solution for human corneal lens and preparation method thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5013714A (en) * | 1988-12-15 | 1991-05-07 | Lindstrom Richard L | Viscoelastic solution |
| US6153582A (en) * | 1998-11-05 | 2000-11-28 | Bausch & Lomb Surgical, Inc. | Defined serumfree medical solution for ophthalmology |
| CN1262866A (en) * | 1999-02-12 | 2000-08-16 | 山东省医学科学院眼科研究所 | Cornea activity preservative fluid |
-
2009
- 2009-03-23 CN CN2009101340052A patent/CN101524063B/en active Active
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102132697A (en) * | 2010-01-21 | 2011-07-27 | 周海华 | Amniotic membrane long-term preserving fluid and preparation method thereof |
| CN105284788A (en) * | 2015-11-20 | 2016-02-03 | 厦门大学 | Cornea metaphase preservation liquid and preparation method thereof |
| CN105284788B (en) * | 2015-11-20 | 2017-11-07 | 厦门大学 | A kind of cornea middle term preserving fluid and preparation method thereof |
| CN106900695A (en) * | 2017-04-01 | 2017-06-30 | 北京焕生汇生物技术研究院 | A kind of mankind's inductive pluripotent stem cells are special to freeze reagent and cryopreservation methods |
| WO2020019516A1 (en) * | 2018-07-26 | 2020-01-30 | 姚晓明 | Cornea medium-term preserving fluid |
| CN111802378A (en) * | 2020-07-24 | 2020-10-23 | 镇江雷音再生医学科技有限公司 | SMILE (Small Scale Integrated Circuit) -derived protective solution for human corneal lens and preparation method thereof |
| CN111802378B (en) * | 2020-07-24 | 2022-02-08 | 镇江雷音再生医学科技有限公司 | SMILE (Small Scale Integrated Circuit) -derived protective solution for human corneal lens and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101524063B (en) | 2012-06-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chhatri et al. | Cryogenic fabrication of savlon loaded macroporous blends of alginate and polyvinyl alcohol (PVA). Swelling, deswelling and antibacterial behaviors | |
| ES2778375T3 (en) | Polymer-based hydrogel | |
| Chirife et al. | In vitro antibacterial activity of concentrated polyethylene glycol 400 solutions | |
| CN101524063B (en) | Cornea mid-term preservation liquid | |
| JP2017522315A (en) | Layered corneal preservation solution | |
| CN105230604A (en) | Marrow mesenchymal stem cell preservation solution and application thereof | |
| RU2184448C2 (en) | Solution to store corneal tissues including hyaluronic acid | |
| CN111407774B (en) | Ophthalmic composition and preparation method thereof | |
| CN103053511A (en) | Cornea metaphase preservation solution, and preparing and using methods thereof | |
| KR101816802B1 (en) | Cryoprotective Agent Containing Exopolysaccharide from Pseudoalteromonas sp. CY01 | |
| US10701928B2 (en) | Method for animal tissue material disinfection and sterilization and corresponding animal tissue soak solution | |
| CN105284788A (en) | Cornea metaphase preservation liquid and preparation method thereof | |
| Giurgola et al. | Antimycotic efficacy and safety of a new cold corneal storage medium by Time–Kill and toxicity studies | |
| CN1270605C (en) | Cornea middle term preserving fluid | |
| CN109258622A (en) | A kind of autoserum cornea middle term preserving fluid for animals and preparation method thereof | |
| CN103190392B (en) | A kind of cornea middle term preserving fluid and preparation method thereof | |
| CN111955455B (en) | Preservation solution for maintaining cornea activity | |
| KR20110104367A (en) | Ophthalmic compositions without preservatives | |
| CN1262866A (en) | Cornea activity preservative fluid | |
| Van Horn et al. | Corneal preservation: recent advances | |
| Ritterband et al. | Efficacy and safety of moxifloxacin as an additive in Optisol-GS a preservation medium for corneal donor tissue | |
| CN111001010B (en) | External eye operation flushing fluid and preparation method thereof | |
| RU2475218C1 (en) | Method for separation and organotypic preservation of allogenic limb transplant | |
| CN112545917A (en) | Disinfecting gel and preparation method thereof | |
| Fong et al. | Cryopreservation of the rabbit cornea: freezing with dimethyl sulphoxide in air or in medium |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230412 Address after: 518000, No. 102, West Ring South Road, Buxin Community, Dapeng Street, Dapeng New District, Shenzhen, Guangdong Province Patentee after: Shenzhen Duanli Biological Group Co.,Ltd. Address before: 518034 Guangdong city of Shenzhen province Futian District honey Lake Road and Nonglin Road intersection of Shenzhen City Eye Hospital Patentee before: Yao Xiaoming |