WO2020019516A1 - 一种角膜中期保存液 - Google Patents

一种角膜中期保存液 Download PDF

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WO2020019516A1
WO2020019516A1 PCT/CN2018/108742 CN2018108742W WO2020019516A1 WO 2020019516 A1 WO2020019516 A1 WO 2020019516A1 CN 2018108742 W CN2018108742 W CN 2018108742W WO 2020019516 A1 WO2020019516 A1 WO 2020019516A1
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preservation solution
corneal
cornea
corneal preservation
preservation
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PCT/CN2018/108742
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English (en)
French (fr)
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姚晓明
王明武
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姚晓明
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0205—Chemical aspects
    • A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0205—Chemical aspects
    • A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
    • A01N1/0215—Disinfecting agents, e.g. antimicrobials for preserving living parts

Definitions

  • the invention relates to a corneal medium-term preservation solution, in particular to a preservation solution for human cornea preservation before transplantation.
  • the current corneal preservation methods can be divided into long-term preservation, medium-term preservation, and short-term preservation according to the length of time.
  • corneal tissue preservation solution is stored at 4 ° C for intermediate-term preservation. This method can not only provide higher-quality cornea, but also Corneal transplant surgery has improved timing.
  • commonly used intermediate preservation solutions include Optisol, K-SOK, etc., which have similar functions but different components, but the protective effects of each preservation solution on the corneal endothelium are also different.
  • the purpose of the present invention is to provide a corneal medium-term preservation solution with better preservation effect and beneficial effect on the corneal endothelium.
  • the technical solution provided by the present invention is as follows:
  • a corneal preservation solution includes the following components at each concentration:
  • the corneal preservation solution includes the following components at various concentrations:
  • the corneal preservation solution further includes a refrigerated component with the following concentration for refrigerated at about 4 ° C:
  • the corneal preservation solution further includes a frozen component of the following concentration for freezing at about -20 ° C:
  • each 100 ml of the corneal medium-term preservation solution further comprises 1 ml of a lipid-concentrated component
  • the lipid-concentrated component includes the components with the following concentrations:
  • the Lipid Concentrate is generally used in the process of culturing suspension cells to produce recombinant proteins in large quantities. This component can promote the healthy growth of cells under serum-free conditions and secrete recombinant proteins. Therefore, compared with other existing preservation Liquid, adding lipid concentrated ingredients is more conducive to preservation of cell survival.
  • Planic F-68 in the lipid concentration component is an inorganic surface-active substance, which can promote the dissolution of the lipid concentration component and also promote the survival of cells in suspension culture.
  • the corneal preservation solution further includes cefazolin.
  • the cefazolin used in the present invention has a broader antibacterial spectrum, covering most Gram-positive and some Gram-negative bacteria.
  • the beneficial effect of the massage is that, compared with the existing preservation solution, the corneal medium-term preservation solution provided by the present invention has better antibacterial effect, better cell preservation effect, and better preservation effect on the corneal endothelium.
  • Free Zinc in the above zinc sulfide 1.4x 10-12M at 36 °C; 1.7x 10-12M at 4 °C; the stock solution is 1mM (used after 1000-fold dilution, 200ul / 200ml), and the pH is adjusted with 1M KOH To about 7.4, add 1M KOH slowly to make the salt completely dissolved.
  • the lipid concentrated ingredients include:

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Dentistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
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Abstract

一种角膜中期保存液，包括如下各组分：K 2SO 4、L-天冬氨酸、硫酸镁、氢氧化钙、磷酸钾、羟乙基哌嗪乙硫磺酸、EDTA、牛磺酸、硫化锌、N-(叔丁基)盐酸羟胺等。有益效果在于，本角膜中期保存液和现有的保存液相比，抗菌效果更好，细胞保存效果更好，特别是对角膜内皮的保护效果更好。

Description

一种角膜中期保存液 技术领域
本发明涉及一种角膜中期保存液，特别涉及一种移植术前，供人体角膜保存的保存液。
背景技术
现在角膜保存的方法，根据时间的长短，可以分为长期保存、中期保存和短期保存，中期保存一般采用角膜片组织保存液4℃保存，这一方法不仅可以提供较高质量的角膜，而且是角膜移植手术在时间安排方面得以改善。目前常用的中期保存液有Optisol、K-SOK等，其功能相似，但是成分不同，但是各个保存液对角膜内皮的保护效果也不尽相同。
发明内容
本发明的目的在于提供一种保存效果更好，有益是对角膜内皮保护效果更好的角膜中期保存液，为实现该目的，本发明提供的技术方案如下：
一种角膜中期保存液，所述的角膜保存液包括如下各浓度的组分：
Figure PCTCN2018108742-appb-000001
进一步的，所述的角膜保存液包括如下各浓度的组分：
Figure PCTCN2018108742-appb-000002
Figure PCTCN2018108742-appb-000003
进一步的，所述的角膜保存液还包括如下浓度的冷藏组分，用于4℃左右冷藏：
葡聚糖            4-6g/100ml
n-乙酰半胱氨酸    0.4-0.6mM。
进一步的，所述的角膜保存液还包括如下浓度的冷冻组分，用于-20℃左右冷冻：
磷酸肌酸         4-6mM。
进一步的，每100ml所述的角膜中期保存液还包括有1ml的脂质浓缩成分，所述的脂质浓缩成分包括有如下各浓度的组分：
Figure PCTCN2018108742-appb-000004
Figure PCTCN2018108742-appb-000005
脂质浓缩成分(The Lipid Concentrate)一般用在培养悬浮细胞以大批生产重组蛋白质的过程中，该成分能促进细胞在无血清条件下的健康生长，分泌重组蛋白质，因而，比起其他现有保存液，加入脂质浓缩成分更有利于保存细胞的生存。
脂质浓缩成分中的普兰尼克F-68是一种无机表面活性物质，可以促进脂质浓缩成分的溶解，同时还有促进悬浮培养细胞存活的作用。
进一步的，所述的角膜保存液还包括有头孢唑林。
本发明采用的头孢唑林具有更广泛的抗菌谱，覆盖大多数革兰氏阳性及一些革兰氏阴性细菌。
本按摩的有益效果在于，本发明提供的角膜中期保存液和现有的保存液相比，抗菌效果更好，细胞保存效果更好，本保存液对角膜内皮的保护效果更好。
具体实施方式
以200ml的角膜中期保存液为例，首先，准备如下各组分的物质：
Figure PCTCN2018108742-appb-000006
Figure PCTCN2018108742-appb-000007
上述硫酸镁中的自由Mg 2+:1.2mM at 4℃；1.17mM at 36℃。
上述氢氧化钙中的自由Ca 2+:1mM at 36℃。
上述硫化锌中的自由Zinc:1.4x 10-12M at 36℃；1.7x 10-12M at 4℃；储备液是1mM(稀释1000倍后使用，200ul/200ml)，同时用1M KOH将pH值调至7.4左右，加1M KOH时要慢，以使盐分彻底溶解。
还准备如下组分：
Figure PCTCN2018108742-appb-000008
Figure PCTCN2018108742-appb-000009
将上述序号1-13的组分按照200mL的用量来混合各物质，在室温下搅拌1h左右即可，然后再将容量加至190ml,再添加1mol/L KOH将pH值调至7.4,不断调整使得最终体积达到200ml，此时测渗透压应该为314mOsmoles，用0.2μ大小的过滤器除菌后，分装到角膜保存瓶中，得到保存液1，然后每100ml保存液1中加1ml的脂质浓缩成分，所述的脂质浓缩成分包括有：
Figure PCTCN2018108742-appb-000010
Figure PCTCN2018108742-appb-000011
最终得到所述的角膜中期保存液。

Claims (6)

  1. 一种角膜中期保存液，其特征在于，所述的角膜保存液包括如下各浓度的组分：
    Figure PCTCN2018108742-appb-100001
  2. 如权利要求1所述的角膜中期保存液，其特征在于，所述的角膜保存液包括如下各浓度的组分：
    Figure PCTCN2018108742-appb-100002
    Figure PCTCN2018108742-appb-100003
  3. 如权利要求1或2所述的角膜中期保存液，其特征在于，所述的角膜保存液还包括如下浓度的冷藏组分：
    葡聚糖           4-6g/100ml
    n-乙酰半胱氨酸   0.4-0.6mM。
  4. 如权利要求1或2所述的角膜中期保存液，其特征在于，所述的角膜保存液还包括如下浓度的冷冻组分：
    磷酸肌酸       4-6mM。
  5. 如权利要求1或2所述的角膜中期保存液，其特征在于，每100ml所述的角膜中期保存液还包括有1ml的脂质浓缩成分，所述的脂质浓缩成分包括有如下各浓度的组分：
    Figure PCTCN2018108742-appb-100004
  6. 如权利要求1或2所述的角膜中期保存液，其特征在于，所述的角膜保存液还包括有头孢唑林。
PCT/CN2018/108742 2018-07-26 2018-09-29 一种角膜中期保存液 WO2020019516A1 (zh)

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CN109329274A (zh) * 2018-11-29 2019-02-15 镇江雷音再生医学科技有限公司 一种含重组人血清白蛋白的角膜中期保存液及其制备方法
CN112514892B (zh) * 2020-12-25 2022-06-14 广州赛莱拉干细胞科技股份有限公司 一种外泌体冻存保护液及其制备方法
CN115777696B (zh) * 2023-02-09 2023-05-30 北京中仪康卫医疗器械有限公司 一种玻璃化冷冻解冻液套装

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CN106135197A (zh) * 2016-07-04 2016-11-23 拜欧迪赛尔（北京）生物科技有限公司 一种无血清成分的眼角膜中期保存液

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US20080131866A1 (en) * 2003-12-17 2008-06-05 The Regents Of The University Of California Tissue Preservation Media
CN101524063A (zh) * 2009-03-23 2009-09-09 姚晓明 角膜中期保存液
CN103053511A (zh) * 2012-12-10 2013-04-24 山东省眼科研究所 一种角膜中期保存液及其制备和使用方法
CN103190392A (zh) * 2013-03-28 2013-07-10 北京大学第三医院 一种角膜中期保存液及其制备方法
CN105284788A (zh) * 2015-11-20 2016-02-03 厦门大学 一种角膜中期保存液及其制备方法
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