CN109329274A - A kind of cornea middle term preserving fluid and preparation method thereof containing recombination human serum albumin - Google Patents

A kind of cornea middle term preserving fluid and preparation method thereof containing recombination human serum albumin Download PDF

Info

Publication number
CN109329274A
CN109329274A CN201811441172.7A CN201811441172A CN109329274A CN 109329274 A CN109329274 A CN 109329274A CN 201811441172 A CN201811441172 A CN 201811441172A CN 109329274 A CN109329274 A CN 109329274A
Authority
CN
China
Prior art keywords
human serum
serum albumin
recombination human
sodium
hcl
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.)
Pending
Application number
CN201811441172.7A
Other languages
Chinese (zh)
Inventor
苏萍
童剑萍
许伟
吴佳宇
肖丹
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhenjiang Leiyin Regenerative Medical Technology Co Ltd
Original Assignee
Zhenjiang Leiyin Regenerative Medical Technology Co Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Zhenjiang Leiyin Regenerative Medical Technology Co Ltd filed Critical Zhenjiang Leiyin Regenerative Medical Technology Co Ltd
Priority to CN201811441172.7A priority Critical patent/CN109329274A/en
Publication of CN109329274A publication Critical patent/CN109329274A/en
Pending legal-status Critical Current

Links

Classifications

    • 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
    • A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Physiology (AREA)
  • Engineering & Computer Science (AREA)
  • Dentistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

The invention discloses a kind of cornea middle term preserving fluid and preparation method thereof containing recombination human serum albumin, the cornea middle term preserving fluid containing recombination human serum albumin contains glutamine, calcium pantothenate, choline chloride, folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl, KCl, NaCl, NaH2PO4·2H2O, glucose, Dextran 40, Sodium Pyruvate, vitamin C, sodium chondroitin sulfate, Sodium Hyaluronate, non-animal source recombination human serum albumin, gentamicin sulphate, sodium bicarbonate, 4- hydroxyethyl piperazineethanesulfonic acid, phenol red indicator, water for injection.It is an advantage of the invention that by the way that non-animal source recombination human serum albumin is added, alternative serum improves endothelial cell survival rate；Furthermore the survival of endothelial cell is ensured using non-animal source recombination human serum albumin collaboration sodium chondroitin sulfate, Sodium Hyaluronate and Dextran 40 compound system, high degree.

Description

A kind of cornea middle term preserving fluid and preparation method thereof containing recombination human serum albumin
Technical field
The present invention relates to the cornea middle term preserving fluid fields containing recombination human serum albumin, more particularly to one kind containing recombination Cornea middle term preserving fluid of human serum albumins and preparation method thereof.
Background technique
For human serum albumins (HSA) as the most abundant albumen of content in human plasma, major function includes: that (1) maintains Solution colloid osmotic pressure and pH value；(2) combination and transport of the important ligand such as lipid, metal ion, amino acid and other factors； (3) anti-oxidation function.
It is a heart-shaped molecule that the x-ray crystal structure of human serum albumins, which discloses it, has 585 amino acid with list Chain link, containing 67% alpha-helix.Human serum albumins structure it is significant wait levy be there are three homologous region city (I, II and III), often A region forms (A detains B) by two subdomains, is each threaded through variable loop connection, shares 35 cysteines in molecule Residue, 17 pairs of disulfide bond form 8 pairs of double disulphide bridges, and constitute 9 rings, and Cys34 is located in the grade seam of about 10 ° of depths, is aoxidizing It is played a crucial role in reduction reaction.
Cornea middle term preserving fluid approved has a K-Sol, CSM, Dexsol at present, Optisol liquid, Optisol-Gs, The products such as 4 DEG C of Chen Medium, Cornea Cold, Eusol-C, LIFE and Cornisol, wherein K-Sol, CSM, Dexsol, Optisol liquid, Optisol-Gs, Chen 4 DEG C of products of Medium, Cornea Cold and LIFE only pass through the U.S. FDA certification, Eusol-C product are authenticated by U.S. FDA and CE.Equal serum-free ingredient in the product of the above external approval listing, It is primarily due to animal derived serum and there is risk in use, it may be containing endotoxin, virus or mycoplasma etc. no Be easy detection and remove external source pollution, in addition, serum exist in the industrial production problem such as downstream process, purifying products, Raw material is variable, product supply respect has limitation and expensive cost.
Summary of the invention
Goal of the invention: in view of the above-mentioned problems, an object of the present invention be to provide it is a kind of containing recombination human serum albumin Cornea middle term preserving fluid, to solve the technical issues of animal derived serum bring external source pollutes.
The second object of the present invention is to provide a kind of preparation side of cornea middle term preserving fluid containing recombination human serum albumin Method.
Technical solution:
A kind of cornea middle term preserving fluid containing recombination human serum albumin, containing glutamine, calcium pantothenate, choline chloride, Folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl, KCl, NaCl, NaH2PO4·2H2O, glucose, Dextran 40, Sodium Pyruvate, vitamin C, sodium chondroitin sulfate, Sodium Hyaluronate, non-animal source recombination human serum albumin, Gentamicin sulphate, sodium bicarbonate, 4- hydroxyethyl piperazineethanesulfonic acid, phenol red indicator, water for injection, wherein water for injection For solvent, the concentration of other each components is respectively as follows: 0.2~0.8g/L of glutamine, 0.001~0.005g/L of calcium pantothenate, chlorination 0.001~0.008g/L of choline, 0.001~0.006g/L of folic acid, 0.002~0.01g/L of inositol, niacinamide 0.001~ 0.006g/L, 0.001~0.008g/L of pyridoxal HCl, 0.0001~0.001g/L of riboflavin, thiamine HCl 0.001 0.05~0.8g/L of~0.008g/L, KCl, 3.0~6.0g/L of NaCl, NaH2PO4·2H20.01~0.5g/L of O, glucose 0.05~2.0g/L, 0.5~2g/L of Dextran 40,0.001~0.5g/L of Sodium Pyruvate, 0.002~2g/L of vitamin C, 15~25g/L of sodium chondroitin sulfate, 0.1~2g/L of Sodium Hyaluronate, 0.1~10g/L of non-animal source recombination human serum albumin, 0.05~0.7g/L of gentamicin sulphate, 10~24mmol/L of sodium bicarbonate, 20~26mmol/L of 4- hydroxyethyl piperazineethanesulfonic acid, 0.01~0.05g/L of phenol red indicator.
Also contain arginine HCl, cystine, histidine HClH in one of the embodiments,2O, different bright ammonia Acid, leucine, lysine HCl, methionine, phenylalanine, threonine, tryptophan, tyrosine, valine, wherein each component Concentration be respectively as follows: 0.12~0.5g/L of arginine HCl, 0.01~0.1g/L of cystine, histidine HClH2O0.02 ~0.2g/L, 0.03~0.28g/L of isoleucine, 0.03~0.28g/L of leucine, 0.05~0.26g/L of lysine HCl, 0.01~0.06g/L of methionine, 0.03~0.13g/L of phenylalanine, 0.02~0.2g/L of threonine, tryptophan 0.005~ 0.05g/L, 0.03~0.2g/L of tyrosine, 0.04~0.3g/L of valine.
In one of the embodiments, the pH value of the cornea middle term preserving fluid containing recombination human serum albumin be 7.2~ 7.6。
A kind of preparation method of the cornea middle term preserving fluid containing recombination human serum albumin, comprising the following steps:
1) each raw material is weighed respectively according to formula ratio；
2) water for injection will be added in Agitation Tank, temperature is controlled to 38~60 DEG C, in whipping process put into glutamine, KCl、NaCl、NaH2PO4·2H2O, glucose, sodium chondroitin sulfate, Sodium Hyaluronate, Dextran 40, Sodium Pyruvate, carbonic acid Hydrogen sodium and 4- hydroxyethyl piperazineethanesulfonic acid, benefit water for injection to the 60~75% of total amount of preparation, and stir 30~120min；
3) feed liquid temperature in Agitation Tank is controlled to 25~40 DEG C, in whipping process into Agitation Tank put into calcium pantothenate, Choline chloride, folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl and vitamin C mend water for injection To the 80~85% of total amount of preparation, and 20~80min is stirred, obtains mixed liquor one；
4) feed liquid temperature in Agitation Tank is controlled to 16~40 DEG C, gentamicin sulphate, after-teeming is added into mixed liquor one 90~95% with water to total amount of preparation are penetrated, and stir 5~40min, obtain mixed liquor two；
5) feed liquid temperature in Agitation Tank is controlled to 16~40 DEG C, phenol red indicator, fluid infusion is added into mixed liquor two Constant volume simultaneously stirs 10~40min, adjusts pH value to 7.2~7.6 with hydrochloric acid and/or sodium hydroxide, obtains intermediate product and prepare liquid；
6) by intermediate product prepare liquid by prefilter carry out pre-filtering processing, after again through PES filter carry out degerming Filter, obtains pre- feed liquid；
7) non-animal source recombination human serum albumin is added in pre- feed liquid, is uniformly mixed, obtained containing recombinant human serum The cornea middle term preserving fluid of albumin.
In one of the embodiments, in step 2), further include in whipping process, by arginine HCl, cystine, Histidine HClH2O, isoleucine, leucine, lysine HCl, methionine, phenylalanine, threonine, tryptophan, junket Propylhomoserin, valine are added in Agitation Tank.
Further comprising the steps of in one of the embodiments, in step 5): obtained intermediate product prepare liquid, are taken Sample detection, detection pH value, osmotic pressure and appearance, detects qualified standard are as follows: pH value is 7.2~7.6, osmotic pressure be 270~ 340mOsmol/kg, appearance color be light rose it is orange, no suspended substance, without visible foreign matters, without precipitating.
Further comprising the steps of in one of the embodiments, in step 6): intermediate product prepare liquid and are carrying out aseptic filtration PES filter is carried out filter core integrity detection, the criterion of acceptability of filter core are as follows: bubble point value >=58.0psi by front and back.
The utility model has the advantages that compared with prior art, it is an advantage of the invention that
1) the above-mentioned cornea middle term preserving fluid containing recombination human serum albumin, first by the way that recombinant human serum albumin egg is added White, alternative serum provides required active factors and nutriment for cornea tissue and endothelial cell, maintains preservation of cornea The survival of endothelial cell in the process, and any exogenous pollution will not be brought；Furthermore use recombination human serum albumin The compound system of sodium chondroitin sulfate, Sodium Hyaluronate and Dextran 40 is cooperateed with, specific mechanism is sodium chondroitin sulfate, transparent Matter acid sodium and Dextran 40 contain a large amount of hydrophilic polar group, can form hydrogen bond association with hydrone, polymer molecule, To make cornea tissue, high molecular polymer and hydrone organically combine；Non-animal source recombination human serum albumin, sulphur The compounding combination of aching and limp ossein sodium, Sodium Hyaluronate, Dextran 40, by mutual synergy, can effectively tie up The original collagen fiber structure of cornea and the transparency are held, and the holding time is longer, protect the survival of endothelial cell as far as possible, The success rate of corneal transplantation is carried out after improving preservation of cornea, while being also provided with and trying hard to keep for the safety of ophthalmology implanted prosthetics Card.
2) preparation method of the above-mentioned cornea middle term preserving fluid containing recombination human serum albumin, due to using fed batch Mode obtains intermediate product and prepares liquid, can make to mix more evenly between each component, and feed liquid in the Agitation Tank of control different phase Temperature plays each component performance preferably, by non-animal source weight after obtained intermediate product preparation liquid progress aseptic filtration Group human serum albumins is added, and the cornea middle term preserving fluid containing recombination human serum albumin enable effectively maintains cornea Original collagen fiber structure and the transparency, and the holding time is longer, can adapt to the demand in market.
Detailed description of the invention
Fig. 1 is that the cornea middle term preserving fluid containing recombination human serum albumin of embodiment 1 is placed with different time after cornea Display image when section under microscope；
Fig. 2 is comparative example 1 without non-animal source recombination human serum albumin, sodium chondroitin sulfate, Sodium Hyaluronate, the right side Revolve the display image when cornea middle term preserving fluid of sugared acid anhydride 40 is placed with different time sections after cornea under microscope.
Specific embodiment
Drug and reagent
Glutamine, calcium pantothenate, choline chloride, folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl、KCl、NaCl、NaH2PO4·2H2O, glucose, Dextran 40, Sodium Pyruvate, vitamin C, sodium chondroitin sulfate, thoroughly Bright matter acid sodium, non-animal source recombination human serum albumin, gentamicin sulphate, sodium bicarbonate, arginine HCl, cystine, group Propylhomoserin HClH2O, isoleucine, leucine, lysine HCl, methionine, phenylalanine, threonine, tryptophan, junket ammonia Acid, valine are bulk pharmaceutical chemicals or pharmaceutic adjuvant rank.
Non-animal source recombination human serum albumin is medical rank, specially Huabei Pharmaceutic Co., Ltd's biotechnology The 201801D2001 lot number product of branch company.Non-animal source recombination human serum albumin be it is fermented by recombinant yeast pichia pastoris, point Safer selection can be provided for serum-free medium without any animal source component from highly purified acquisition.With tire ox blood (FBS), blood plasma source albumin (pHSA) are compared with bovine serum albumin(BSA) (BSA) clearly, non-animal source recombination human serum albumin With higher purity and the consistent structure of human serum albumins, better lot stability；Meanwhile non-animal source recombination weight Group human serum albumins is also widely used as auxiliary material, excipient, stabilizer and embedding medium etc. in biological medicine production.
Embodiment 1
A kind of cornea middle term preserving fluid containing recombination human serum albumin, containing glutamine, calcium pantothenate, choline chloride, Folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl, KCl, NaCl, NaH2PO4·2H2O, glucose, Dextran 40, Sodium Pyruvate, vitamin C, sodium chondroitin sulfate, Sodium Hyaluronate, non-animal source recombination human serum albumin, Gentamicin sulphate, sodium bicarbonate, 4- hydroxyethyl piperazineethanesulfonic acid, phenol red indicator, water for injection, arginine HCl, Guang Propylhomoserin, histidine HClH2O, isoleucine, leucine, lysine HCl, methionine, phenylalanine, threonine, color ammonia Acid, tyrosine, valine, wherein water for injection is solvent, and the concentration of other each components is respectively as follows: glutamine 0.6g/L, general Sour calcium 0.002g/L, choline chloride 0.002g/L, folic acid 0.002g/L, inositol 0.004g/L, niacinamide 0.001g/L, pyrrole are trembled Aldehyde HCl 0.002g/L, riboflavin 0.0002g/L, thiamine HCl 0.002g/L, KCl 0.4g/L, NaCl 4.8g/L, NaH2PO4·2H2O 0.14g/L, glucose 0.5g/L, Dextran 40 1.2g/L, Sodium Pyruvate 0.04g/L, vitamin C 0.01g/L, sodium chondroitin sulfate 22g/L, Sodium Hyaluronate 0.5g/L, non-animal source recombination human serum albumin 0.5g/L, sulphur Sour gentamicin 0.4g/L, sodium bicarbonate 16mmol/L, 4- hydroxyethyl piperazineethanesulfonic acid 25mmol/L, phenol red indicator 0.015g/L, arginine HCl0.25g/L, cystine 0.048g/L, histidine HClH2O 0.084g/L, isoleucine 0.104g/L, leucine 0.104g/L, lysine HCl 0.146g/L, methionine 0.03g/L, phenylalanine 0.066g/L, Threonine 0.096g/L, tryptophan 0.02g/L, tyrosine 0.072g/L, valine 0.094g/L.Wherein, contain recombinant human serum The pH value of the cornea middle term preserving fluid of albumin is 7.3.
A kind of preparation method of the cornea middle term preserving fluid containing recombination human serum albumin, comprising the following steps:
1) each raw material is weighed respectively according to formula ratio；
2) water for injection will be added in Agitation Tank, temperature is controlled to 60 DEG C, in whipping process put into glutamine, KCl, NaCl、NaH2PO4·2H2O, glucose, sodium chondroitin sulfate, Sodium Hyaluronate, Dextran 40, Sodium Pyruvate, bicarbonate Sodium, arginine HCl, cystine, histidine HClH2O, isoleucine, leucine, relies 4- hydroxyethyl piperazineethanesulfonic acid Propylhomoserin HCl, methionine, phenylalanine, threonine, tryptophan, tyrosine, valine mend water for injection to total amount of preparation 60%, and stir 40min；
3) feed liquid temperature in Agitation Tank is controlled to 40 DEG C, puts into calcium pantothenate, chlorination into Agitation Tank in whipping process Choline, folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl and vitamin C mend water for injection to total The 80% of amount of preparation, and 30min is stirred, obtain mixed liquor one；
4) feed liquid temperature in Agitation Tank is controlled to 40 DEG C, gentamicin sulphate is added into mixed liquor one, mend injection Water stirs 10min to the 90% of total amount of preparation, obtains mixed liquor two；
5) feed liquid temperature in Agitation Tank is controlled to 40 DEG C, phenol red indicator, fluid infusion constant volume is added into mixed liquor two And 15min is stirred, pH value is adjusted to 7.3 with hydrochloric acid and/or sodium hydroxide, is obtained intermediate product and is prepared liquid；
Wherein, the intermediate product obtained prepare liquid, are sampled detection, and detection pH value, osmotic pressure and appearance detect qualification Standard are as follows: pH value is 7.2~7.6, osmotic pressure is 270~340mOsmol/kg, appearance color is that light rose is orange, no suspension Object, without visible foreign matters, without precipitating.
6) by intermediate product prepare liquid by prefilter carry out pre-filtering processing, after again through PES filter carry out degerming Filter, obtains pre- feed liquid；
Wherein, intermediate product prepare liquid before and after carrying out aseptic filtration, and PES filter is carried out filter core integrity detection, filter The criterion of acceptability of core are as follows: bubble point value >=58.0psi.
7) non-animal source recombination human serum albumin is added in pre- feed liquid, is uniformly mixed, obtained containing recombinant human serum The cornea middle term preserving fluid of albumin carries out filling afterwards.
Embodiment 2
A kind of cornea middle term preserving fluid containing recombination human serum albumin, containing glutamine, calcium pantothenate, choline chloride, Folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl, KCl, NaCl, NaH2PO4·2H2O, glucose, Dextran 40, Sodium Pyruvate, vitamin C, sodium chondroitin sulfate, Sodium Hyaluronate, non-animal source recombination human serum albumin, Gentamicin sulphate, sodium bicarbonate, 4- hydroxyethyl piperazineethanesulfonic acid, phenol red indicator, water for injection, arginine HCl, Guang Propylhomoserin, histidine HClH2O, isoleucine, leucine, lysine HCl, methionine, phenylalanine, threonine, color ammonia Acid, tyrosine, valine, wherein water for injection is solvent, and the concentration of other each components is respectively as follows: glutamine 0.3g/L, general Sour calcium 0.002g/L, choline chloride 0.002g/L, folic acid 0.002g/L, inositol 0.004g/L, niacinamide 0.001g/L, pyrrole are trembled Aldehyde HCl 0.002g/L, riboflavin 0.0002g/L, thiamine HCl 0.002g/L, KCl 0.2g/L, NaCl 5.0g/L, NaH2PO4·2H2O 0.2g/L, glucose 1.5g/L, Dextran 40 1g/L, Sodium Pyruvate 0.2g/L, vitamin C 0.1g/ L, sodium chondroitin sulfate 20g/L, Sodium Hyaluronate 0.4g/L, non-animal source recombination human serum albumin 1.0g/L, sulfuric acid celebrating are big Mycin 0.4g/L, sodium bicarbonate 16mmol/L, 4- hydroxyethyl piperazineethanesulfonic acid 25mmol/L, phenol red indicator 0.03g/L, essence Propylhomoserin HCl
0.125g/L, cystine 0.024g/L, histidine HClH2It is O 0.044g/L, isoleucine 0.06g/L, bright Propylhomoserin 0.06g/L, lysine HCl 0.08g/L, methionine 0.02g/L, phenylalanine 0.04g/L, threonine 0.05g/L, Tryptophan 0.01g/L, tyrosine 0.04g/L, valine 0.05g/L.Wherein, the cornea for medium period containing recombination human serum albumin is protected The pH value of liquid storage is 7.3.
A kind of preparation method of the cornea middle term preserving fluid containing recombination human serum albumin, comprising the following steps:
1) each raw material is weighed respectively according to formula ratio；
2) water for injection will be added in Agitation Tank, temperature is controlled to 50 DEG C, in whipping process put into glutamine, KCl, NaCl、NaH2PO4·2H2O, glucose, sodium chondroitin sulfate, Sodium Hyaluronate, Dextran 40, Sodium Pyruvate, bicarbonate Sodium, arginine HCl, cystine, histidine HClH2O, isoleucine, leucine, relies 4- hydroxyethyl piperazineethanesulfonic acid Propylhomoserin HCl, methionine, phenylalanine, threonine, tryptophan, tyrosine, valine mend water for injection to total amount of preparation 70%, and stir 80min；
3) feed liquid temperature in Agitation Tank is controlled to 35 DEG C, puts into calcium pantothenate, chlorination into Agitation Tank in whipping process Choline, folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl and vitamin C mend water for injection to total The 85% of amount of preparation, and 40min is stirred, obtain mixed liquor one；
4) feed liquid temperature in Agitation Tank is controlled to 36 DEG C, gentamicin sulphate is added into mixed liquor one, mend injection Water stirs 15min to the 95% of total amount of preparation, obtains mixed liquor two；
5) feed liquid temperature in Agitation Tank is controlled to 35 DEG C, phenol red indicator, fluid infusion constant volume is added into mixed liquor two And 20min is stirred, pH value is adjusted to 7.3 with hydrochloric acid and/or sodium hydroxide, is obtained intermediate product and is prepared liquid；
Wherein, the intermediate product obtained prepare liquid, are sampled detection, and detection pH value, osmotic pressure and appearance detect qualification Standard are as follows: pH value is 7.2~7.6, osmotic pressure is 270~340mOsmol/kg, appearance color is that light rose is orange, no suspension Object, without visible foreign matters, without precipitating.
6) by intermediate product prepare liquid by prefilter carry out pre-filtering processing, after again through PES filter carry out degerming Filter, obtains pre- feed liquid；
Wherein, intermediate product prepare liquid before and after carrying out aseptic filtration, and PES filter is carried out filter core integrity detection, filter The criterion of acceptability of core are as follows: bubble point value >=58.0psi.
7) non-animal source recombination human serum albumin is added in pre- feed liquid, is uniformly mixed, obtained containing recombinant human serum The cornea middle term preserving fluid of albumin carries out filling afterwards.
Embodiment 3
A kind of cornea middle term preserving fluid containing recombination human serum albumin, containing glutamine, calcium pantothenate, choline chloride, Folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl, KCl, NaCl, NaH2PO4·2H2O, glucose, Dextran 40, Sodium Pyruvate, vitamin C, sodium chondroitin sulfate, Sodium Hyaluronate, non-animal source recombination human serum albumin, Gentamicin sulphate, sodium bicarbonate, 4- hydroxyethyl piperazineethanesulfonic acid, phenol red indicator, water for injection, arginine HCl, Guang Propylhomoserin, histidine HClH2O, isoleucine, leucine, lysine HCl, methionine, phenylalanine, threonine, color ammonia Acid, tyrosine, valine, wherein water for injection is solvent, and the concentration of other each components is respectively as follows: glutamine 0.6g/L, general Sour calcium 0.004g/L, choline chloride 0.004g/L, folic acid 0.004g/L, inositol 0.008g/L, niacinamide 0.002g/L, pyrrole are trembled Aldehyde HCl 0.004g/L, riboflavin 0.0004g/L, thiamine HCl 0.004g/L, KCl 0.2g/L, NaCl 4.8g/L, NaH2PO4·2H2O 0.14g/L, glucose 1g/L, Dextran 40 1g/L, Sodium Pyruvate 0.15g/L, vitamin C 0.0081g/L, sodium chondroitin sulfate 22g/L, Sodium Hyaluronate 0.2g/L, non-animal source recombination human serum albumin 1g/L, sulphur Sour gentamicin 0.4g/L, sodium bicarbonate 16mmol/L, 4- hydroxyethyl piperazineethanesulfonic acid 25mmol/L, phenol red indicator 0.05g/L, arginine HCl0.5g/L, cystine 0.09g/L, histidine HClH2O 0.16g/L, isoleucine 0.2g/L, leucine 0.2g/L, lysine HCl 0.29g/L, methionine 0.06g/L, phenylalanine 0.132g/L, threonine 0.192g/L, tryptophan 0.04g/L, tyrosine 0.144g/L, valine 0.188g/L.Wherein, contain recombination human serum albumin Cornea middle term preserving fluid pH value be 7.4.
A kind of preparation method of the cornea middle term preserving fluid containing recombination human serum albumin, comprising the following steps:
1) each raw material is weighed respectively according to formula ratio；
2) water for injection will be added in Agitation Tank, temperature is controlled to 40 DEG C, in whipping process put into glutamine, KCl, NaCl、NaH2PO4·2H2O, glucose, sodium chondroitin sulfate, Sodium Hyaluronate, Dextran 40, Sodium Pyruvate, bicarbonate Sodium, arginine HCl, cystine, histidine HClH2O, isoleucine, leucine, relies 4- hydroxyethyl piperazineethanesulfonic acid Propylhomoserin HCl, methionine, phenylalanine, threonine, tryptophan, tyrosine, valine mend water for injection to total amount of preparation 75%, and stir 100min；
3) feed liquid temperature in Agitation Tank is controlled to 40 DEG C, puts into calcium pantothenate, chlorination into Agitation Tank in whipping process Choline, folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl and vitamin C mend water for injection to total The 82% of amount of preparation, and 40min is stirred, obtain mixed liquor one；
4) feed liquid temperature in Agitation Tank is controlled to 35 DEG C, gentamicin sulphate is added into mixed liquor one, mend injection Water stirs 20min to the 92% of total amount of preparation, obtains mixed liquor two；
5) feed liquid temperature in Agitation Tank is controlled to 36 DEG C, phenol red indicator, fluid infusion constant volume is added into mixed liquor two And 25min is stirred, pH value is adjusted to 7.4 with hydrochloric acid and/or sodium hydroxide, is obtained intermediate product and is prepared liquid；
Wherein, the intermediate product obtained prepare liquid, are sampled detection, and detection pH value, osmotic pressure and appearance detect qualification Standard are as follows: pH value is 7.2~7.6, osmotic pressure is 270~340mOsmol/kg, appearance color is that light rose is orange, no suspension Object, without visible foreign matters, without precipitating.
6) by intermediate product prepare liquid by prefilter carry out pre-filtering processing, after again through PES filter carry out degerming Filter, obtains pre- feed liquid；
Wherein, intermediate product prepare liquid before and after carrying out aseptic filtration, and PES filter is carried out filter core integrity detection, filter The criterion of acceptability of core are as follows: bubble point value >=58.0psi.
7) non-animal source recombination human serum albumin is added in pre- feed liquid, is uniformly mixed, obtained containing recombinant human serum The cornea middle term preserving fluid of albumin carries out filling afterwards.
Embodiment 4
A kind of cornea middle term preserving fluid containing recombination human serum albumin, containing glutamine, calcium pantothenate, choline chloride, Folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl, KCl, NaCl, NaH2PO4·2H2O, glucose, Dextran 40, Sodium Pyruvate, vitamin C, sodium chondroitin sulfate, Sodium Hyaluronate, non-animal source recombination human serum albumin, Gentamicin sulphate, sodium bicarbonate, 4- hydroxyethyl piperazineethanesulfonic acid, phenol red indicator, water for injection, wherein water for injection For solvent, the concentration of other each components be respectively as follows: glutamine 0.3g/L, calcium pantothenate 0.002g/L, choline chloride 0.002g/L, Folic acid 0.002g/L, inositol 0.004g/L, niacinamide 0.001g/L, pyridoxal HCl 0.002g/L, riboflavin 0.0002g/ L, thiamine HCl 0.002g/L, KCl 0.2g/L, NaCl5.0g/L, NaH2PO4·2H2O 0.2g/L, glucose 1.5g/ L, Dextran 40 1g/L, Sodium Pyruvate 0.2g/L, vitamin C 0.1g/L, sodium chondroitin sulfate 20g/L, Sodium Hyaluronate 0.4g/L, non-animal source recombination human serum albumin 1.0g/L, gentamicin sulphate 0.4g/L, sodium bicarbonate 16mmol/L, 4- Hydroxyethyl piperazineethanesulfonic acid 25mmol/L, phenol red indicator 0.05g/L.
Wherein, the pH value of the cornea middle term preserving fluid containing recombination human serum albumin is 7.4.
A kind of preparation method of the cornea middle term preserving fluid containing recombination human serum albumin, comprising the following steps:
1) each raw material is weighed respectively according to formula ratio；
2) water for injection will be added in Agitation Tank, temperature is controlled to 60 DEG C, in whipping process put into glutamine, KCl, NaCl、NaH2PO4·2H2O, glucose, sodium chondroitin sulfate, Sodium Hyaluronate, Dextran 40, Sodium Pyruvate, sodium bicarbonate With 4- hydroxyethyl piperazineethanesulfonic acid, fluid infusion water for injection to the 60% of total injection water, and 40min is stirred；
3) feed liquid temperature in Agitation Tank is controlled to 40 DEG C, puts into calcium pantothenate, chlorination into Agitation Tank in whipping process Choline, folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl and vitamin C mend water for injection to total The 80% of amount of preparation, and 30min is stirred, obtain mixed liquor one；
4) feed liquid temperature in Agitation Tank is controlled to 40 DEG C, gentamicin sulphate is added into mixed liquor one, mend injection Water stirs 10min to the 90% of total amount of preparation, obtains mixed liquor two；
5) feed liquid temperature in Agitation Tank is controlled to 40 DEG C, phenol red indicator, fluid infusion constant volume is added into mixed liquor two And 15min is stirred, pH value is adjusted to 7.4 with hydrochloric acid and/or sodium hydroxide, is obtained intermediate product and is prepared liquid；
6) by intermediate product prepare liquid by prefilter carry out pre-filtering processing, after again through PES filter carry out degerming Filter, obtains pre- feed liquid；
7) the pre- solution of non-animal source recombination human serum albumin is added in pre- feed liquid, is uniformly mixed, obtained containing recombination The cornea middle term preserving fluid of human serum albumins carries out filling afterwards.
Comparative example 1
This comparative example 1 and the distinctive points of embodiment 1 are, in this comparative example 1, do not contain sodium chondroitin sulfate, hyalomitome Sour sodium, non-animal source recombination human serum albumin and Dextran 40.
Performance test
The preservation effect for selecting cornea middle term preserving fluid of the porcine cornea verifying containing recombination human serum albumin, saves step such as Under:
1) pig eyeball is won at slaughterhouse scene, is rinsed with antibiotic sterile phosphate buffer.
2) eyeball sclera portion is wrapped up with the wet yarn block containing antibiotic solution, to maintain to save the saturated with moisture environment in bottle Cornea is put into eyeball bottle upwardly, drips antibiotic solution to cornea face, cover tightly by (gauze piece must not contact cornea)；
3) it is put into the transport case of ice cube, it need to be every avoiding ice cube temperature is too low from making cornea with material between eyeball bottle and ice cube Frostbite.Temperature maintains 2~8 DEG C.
4) after transporting laboratory back, eyeball is taken out on superclean bench, pincers is fixed, with 10mm trepan in anterior corneal surface It rotates left and right, does vertical drilling；After confirmation corneoscleral junction is kept completely separate with choroid below, the sclera of corneal film is clamped Edge gently takes out.It is rinsed with antibiotic sterile phosphate buffer.
5) corneal film after flushing is put into the vial of the cornea middle term preserving fluid containing recombination human serum albumin, completely It is immersed in the cornea middle term preserving fluid containing recombination human serum albumin, immediately bottle cover for locking, after posting label, is put into 2~8 DEG C of ice It is saved in case.
Respectively by the cornea middle term preserving fluid of the recombination human serum albumin containing non-animal source of embodiment 1, as control The angle without non-animal source recombination human serum albumin, sodium chondroitin sulfate, Sodium Hyaluronate, Dextran 40 of comparative example 1 Film mid-term preservation liquid saves under the conditions of 4 DEG C respectively, samples respectively at 0 day, 7 days and 14 days, carries out trypan blue-alizarin red joint Dyeing detection endothelial cell survival rate, the result is shown in Figure 1, Fig. 2 and table 1.
Ultrasonic pachymetry number counts in the cornea middle term preserving fluid of 1 embodiment 1 of table and comparative example 1
In Fig. 1, Fig. 1 (a) is the cornea for medium period guarantor containing recombination human serum albumin that fresh porcine cornea is placed into embodiment 1 State diagram when liquid storage；Fig. 1 (b) is the cornea for medium period preservation containing recombination human serum albumin that porcine cornea is placed into embodiment 1 State diagram in liquid after 7 days；Fig. 1 (c) is the cornea for medium period guarantor containing recombination human serum albumin that porcine cornea is placed into embodiment 1 State diagram in liquid storage after 14 days；
Fig. 2 (a) be porcine cornea be placed into comparative example 1 without non-animal source recombination human serum albumin, chondroitin sulfate Sodium, Sodium Hyaluronate, Dextran 40 cornea middle term preserving fluid in state diagram after 7 days；Fig. 2 (b) is placed into for porcine cornea The angle without non-animal source recombination human serum albumin, sodium chondroitin sulfate, Sodium Hyaluronate, Dextran 40 of comparative example 1 State diagram in film mid-term preservation liquid after 14 days；
By Fig. 1 (a) it is found that fresh cornea dyeing corneal endothelial damage following cellular morphology keeps hexagonal structure complete substantially, carefully Intercellular is completely embedded, without obvious cellular swelling, missing and cell membrane fusion, seedless blue dye.
By Fig. 1 and table 1 it is found that using the cornea middle term preserving fluid of the invention containing recombination human serum albumin, porcine cornea When saving 7 days, endothelial cell form rule, arrangement is close, and no obvious tumefaction, nuclear staining is few, and cellular damage is unobvious；It protects When depositing 14 days, endothelial cell form rule, arrangement is close, and nuclear staining slightly increases, and illustrates of the invention containing recombinant human The cornea middle term preserving fluid of pure albumen can effectively maintain the original collagen fiber structure of cornea and the transparency, and holding time It is longer, the survival of endothelial cell is protected as far as possible, improves the success rate of progress corneal transplantation after preservation of cornea, while Safety for ophthalmology implanted prosthetics provides strong guarantee.
By Fig. 2 and table 1 it is found that when porcine cornea saves 7 days in the cornea middle term preserving fluid of comparative example 1, angle in comparative example 1 Film endothelial cell obvious tumefaction, missing, nuclear staining are more, and when saving 14 days, endothelial cell connection relaxation, nuclear staining is more, ties Structure is substantially unclear.

Claims (7)

1. a kind of cornea middle term preserving fluid containing recombination human serum albumin, which is characterized in that containing glutamine, calcium pantothenate, Choline chloride, folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl, KCl, NaCl, NaH2PO4· 2H2O, glucose, Dextran 40, Sodium Pyruvate, vitamin C, sodium chondroitin sulfate, Sodium Hyaluronate, non-animal source recombination Human serum albumins, gentamicin sulphate, sodium bicarbonate, 4- hydroxyethyl piperazineethanesulfonic acid, phenol red indicator, water for injection, Wherein, water for injection is solvent, the concentration of other each components be respectively as follows: 0.2~0.8g/L of glutamine, calcium pantothenate 0.001~ 0.005g/L, 0.001~0.008g/L of choline chloride, 0.001~0.006g/L of folic acid, 0.002~0.01g/L of inositol, nicotinoyl 0.001~0.006g/L of amine, 0.001~0.008g/L of pyridoxal HCl, 0.0001~0.001g/L of riboflavin, thiamine 0.001~0.008g/L of HCl, 0.05~0.8g/L of KCl, 3.0~6.0g/L of NaCl, NaH2PO4·2H2O 0.01~ 0.5g/L, 0.05~2.0g/L of glucose, 0.5~2g/L of Dextran 40,0.001~0.5g/L of Sodium Pyruvate, vitamin C 0.002~2g/L, 15~25g/L of sodium chondroitin sulfate, 0.1~2g/L of Sodium Hyaluronate, non-animal source recombinant human serum albumin egg White 0.1~10g/L, 0.05~0.7g/L of gentamicin sulphate, 10~24mmol/L of sodium bicarbonate, 4- hydroxyethyl piperazineethanesulfonic acid 20~26mmol/L, 0.01~0.05g/L of phenol red indicator.
2. a kind of cornea middle term preserving fluid containing recombination human serum albumin according to claim 1, which is characterized in that also Contain arginine HCl, cystine, histidine HClH2O, isoleucine, leucine, lysine HCl, methionine, benzene Alanine, threonine, tryptophan, tyrosine, valine, wherein the concentration of each component be respectively as follows: arginine HCl 0.12~ 0.5g/L, 0.01~0.1g/L of cystine, histidine HClH20.02~0.2g/L of O, 0.03~0.28g/ of isoleucine L, 0.03~0.28g/L of leucine, 0.05~0.26g/L of lysine HCl, 0.01~0.06g/L of methionine, phenylalanine 0.03~0.13g/L, 0.02~0.2g/L of threonine, 0.005~0.05g/L of tryptophan, 0.03~0.2g/L of tyrosine, figured silk fabrics ammonia 0.04~0.3g/L of acid.
3. a kind of cornea middle term preserving fluid containing recombination human serum albumin according to claim 1, which is characterized in that contain The pH value of the cornea middle term preserving fluid of recombination human serum albumin is 7.2~7.6.
4. a kind of preparation method of the cornea middle term preserving fluid according to claim 1 containing recombination human serum albumin, It is characterized in that, comprising the following steps:
1) each raw material is weighed respectively according to formula ratio；
2) water for injection will be added in Agitation Tank, temperature is controlled to 38~60 DEG C, in whipping process put into glutamine, KCl, NaCl、NaH2PO4·2H2O, glucose, sodium chondroitin sulfate, Sodium Hyaluronate, Dextran 40, Sodium Pyruvate, sodium bicarbonate With 4- hydroxyethyl piperazineethanesulfonic acid, benefit water for injection to the 60~75% of total amount of preparation, and 30~120min of stirring；
3) feed liquid temperature in Agitation Tank is controlled to 25~40 DEG C, puts into calcium pantothenate, chlorination into Agitation Tank in whipping process Choline, folic acid, inositol, niacinamide, pyridoxal HCl, riboflavin, thiamine HCl and vitamin C mend water for injection to total The 80~85% of amount of preparation, and 20~80min is stirred, obtain mixed liquor one；
4) feed liquid temperature in Agitation Tank is controlled to 16~40 DEG C, gentamicin sulphate is added into mixed liquor one, mend injection Water stirs 5~40min to the 90~95% of total amount of preparation, obtains mixed liquor two；
5) feed liquid temperature in Agitation Tank is controlled to 16~40 DEG C, phenol red indicator, fluid infusion constant volume is added into mixed liquor two And 10~40min is stirred, pH value is adjusted to 7.2~7.6 with hydrochloric acid and/or sodium hydroxide, is obtained intermediate product and is prepared liquid；
6) by intermediate product prepare liquid by prefilter carry out pre-filtering processing, after again through PES filter carry out aseptic filtration, obtain To pre- feed liquid；
7) non-animal source recombination human serum albumin is added in pre- feed liquid, is uniformly mixed, obtains egg containing recombinant human serum albumin White cornea middle term preserving fluid.
5. a kind of preparation method of cornea middle term preserving fluid containing recombination human serum albumin according to claim 4, It is characterized in that, further includes in whipping process, by arginine HCl, cystine, histidine HClH2O, different in step 2) Leucine, leucine, lysine HCl, methionine, phenylalanine, threonine, tryptophan, tyrosine, valine are added to and match In flow container.
6. a kind of preparation method of cornea middle term preserving fluid containing recombination human serum albumin according to claim 4, It is characterized in that, further comprising the steps of in step 5): obtained intermediate product prepare liquid, are sampled detection, and detection pH value is seeped Pressure and appearance thoroughly detect qualified standard are as follows: pH value is 7.2~7.6, osmotic pressure is 270~340mOsmol/kg, appearance color It is orange for light rose, no suspended substance, without visible foreign matters, without precipitating.
7. a kind of preparation method of cornea middle term preserving fluid containing recombination human serum albumin according to claim 4, Be characterized in that, further comprising the steps of in step 6): intermediate product prepare liquid before and after carrying out aseptic filtration, by PES filter into Row filter core integrity detection, the criterion of acceptability of filter core are as follows: bubble point value >=58.0psi.
CN201811441172.7A 2018-11-29 2018-11-29 A kind of cornea middle term preserving fluid and preparation method thereof containing recombination human serum albumin Pending CN109329274A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201811441172.7A CN109329274A (en) 2018-11-29 2018-11-29 A kind of cornea middle term preserving fluid and preparation method thereof containing recombination human serum albumin

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201811441172.7A CN109329274A (en) 2018-11-29 2018-11-29 A kind of cornea middle term preserving fluid and preparation method thereof containing recombination human serum albumin

Publications (1)

Publication Number Publication Date
CN109329274A true CN109329274A (en) 2019-02-15

Family

ID=65319388

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201811441172.7A Pending CN109329274A (en) 2018-11-29 2018-11-29 A kind of cornea middle term preserving fluid and preparation method thereof containing recombination human serum albumin

Country Status (1)

Country Link
CN (1) CN109329274A (en)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110476954A (en) * 2019-09-11 2019-11-22 广东博与再生医学有限公司 Collagen tissue material preservation liquid and its application
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
WO2023282338A1 (en) * 2021-07-07 2023-01-12 株式会社セルージョン Cell seeding agent and substrate for cell transplantation use
WO2023190941A1 (en) * 2022-03-31 2023-10-05 学校法人同志社 Corneal endothelial cell preparation containing no culture-derived component, and method for producing same
EP4368021A1 (en) * 2022-11-10 2024-05-15 AL.CHI.MI.A. S.r.l. Preservation solution, preservation system and method for preserving bioological tissues in vitro, in particular corneal tissues
EP4368020A1 (en) * 2022-11-10 2024-05-15 AL.CHI.MI.A. S.r.l. Preservation solution, preservation system and method for peserving bioological tissues
JP7504518B2 (en) 2021-07-07 2024-06-24 株式会社セルージョン Cell seeding agent and cell transplantation substrate

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101502257A (en) * 2009-03-13 2009-08-12 厦门大学 Corneal midterm preservation solution and preparation method thereof
CN105284788A (en) * 2015-11-20 2016-02-03 厦门大学 Cornea metaphase preservation liquid and preparation method thereof
CN106135197A (en) * 2016-07-04 2016-11-23 拜欧迪赛尔（北京）生物科技有限公司 A kind of cornea mid-term preservation liquid of serum-free composition
US20170203002A1 (en) * 2013-06-25 2017-07-20 James H. FORSELL Method for preparing mammalian tissue for storage, implant, and transplant
CN108849858A (en) * 2018-07-26 2018-11-23 姚晓明 A kind of cornea middle term preserving fluid

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101502257A (en) * 2009-03-13 2009-08-12 厦门大学 Corneal midterm preservation solution and preparation method thereof
US20170203002A1 (en) * 2013-06-25 2017-07-20 James H. FORSELL Method for preparing mammalian tissue for storage, implant, and transplant
CN105284788A (en) * 2015-11-20 2016-02-03 厦门大学 Cornea metaphase preservation liquid and preparation method thereof
CN106135197A (en) * 2016-07-04 2016-11-23 拜欧迪赛尔（北京）生物科技有限公司 A kind of cornea mid-term preservation liquid of serum-free composition
CN108849858A (en) * 2018-07-26 2018-11-23 姚晓明 A kind of cornea middle term preserving fluid

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
李青旺主编: "《动物细胞工程与实践》", 30 June 2005, 化学工业出版社教材出版中心 *
杨扬等: "自制人血清中期角膜保存液对兔角膜的保存效果", 《第三军医大学学报》 *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110476954A (en) * 2019-09-11 2019-11-22 广东博与再生医学有限公司 Collagen tissue material preservation liquid and its application
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
WO2023282338A1 (en) * 2021-07-07 2023-01-12 株式会社セルージョン Cell seeding agent and substrate for cell transplantation use
JP7504518B2 (en) 2021-07-07 2024-06-24 株式会社セルージョン Cell seeding agent and cell transplantation substrate
WO2023190941A1 (en) * 2022-03-31 2023-10-05 学校法人同志社 Corneal endothelial cell preparation containing no culture-derived component, and method for producing same
EP4368021A1 (en) * 2022-11-10 2024-05-15 AL.CHI.MI.A. S.r.l. Preservation solution, preservation system and method for preserving bioological tissues in vitro, in particular corneal tissues
EP4368020A1 (en) * 2022-11-10 2024-05-15 AL.CHI.MI.A. S.r.l. Preservation solution, preservation system and method for peserving bioological tissues

Similar Documents

Publication Publication Date Title
CN109329274A (en) A kind of cornea middle term preserving fluid and preparation method thereof containing recombination human serum albumin
US4443432A (en) Ophthmalic irrigating solution
US4550022A (en) Tissue irrigating solution
NO328567B1 (en) Acidic insulin preparations with improved stability
US20100209532A1 (en) Targeted oxygen delivery via intravenous or intra-arterial infusion of oxygenated polymerized hemoglobin solutions
CN1367703A (en) Freeze dried HGF preparations
CN110563836A (en) preparation method of cross-linked hemoglobin and organ perfusate containing hemoglobin
CN110495450B (en) Cell cryopreservation transport solution and application thereof
WO2002045508A1 (en) Use of alpha-keratose as a blood plasma expander
US4837021A (en) Two part tissue irrigating solution
CN105288627A (en) Cardioplegic solution with oxygen carrying and supplying function and preparation method and application of cardioplegic solution
CN114146163A (en) Preparation method of semaglutide preparation
ZA200504202B (en) Prekallikrein depleted plasma derived albumin fraction.
CN104189891A (en) Recombinant human erythropoietin preparation without human albumin
CN111657269B (en) Method for protecting and treating fibrin glue before preservation of SMILE-derived human corneal lens
US11077193B2 (en) Nerve growth factor composition and powder injection
CN105543167B (en) The preservation and transportation resources of stem cell
EP4368020A1 (en) Preservation solution, preservation system and method for peserving bioological tissues
CN103800323A (en) Compound amino acid injection solution 18AA-V composition and preparation method thereof
RU2815501C1 (en) Solution for pre-transplant preparation of donor lungs
Li et al. Application of polymerized porcine hemoglobin in the ex vivo normothermic machine perfusion of rat livers
CN111802378B (en) SMILE (Small Scale Integrated Circuit) -derived protective solution for human corneal lens and preparation method thereof
EP4368021A1 (en) Preservation solution, preservation system and method for preserving bioological tissues in vitro, in particular corneal tissues
ES2893239T3 (en) Composition of nerve growth factor and injection powder
US20180110833A1 (en) Nerve growth factor composition and powder injection

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20190215