CN110810397A - Hematopoietic stem cell cryopreservation liquid and preparation method and application method thereof - Google Patents
Hematopoietic stem cell cryopreservation liquid and preparation method and application method thereof Download PDFInfo
- Publication number
- CN110810397A CN110810397A CN201810901210.6A CN201810901210A CN110810397A CN 110810397 A CN110810397 A CN 110810397A CN 201810901210 A CN201810901210 A CN 201810901210A CN 110810397 A CN110810397 A CN 110810397A
- Authority
- CN
- China
- Prior art keywords
- myosin
- hematopoietic stem
- stem cell
- frozen stock
- stock solution
- 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
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
- A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
-
- 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)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Biophysics (AREA)
- Physiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to a hematopoietic stem cell frozen stock solution and a preparation method and a using method thereof, wherein the hematopoietic stem cell frozen stock solution comprises lecithin, ovalbumin, myosin, platelet lysate and buffer solution; wherein, in every 100ml of the frozen stock solution, the mass range of the lecithin is 1g to 4g, the mass range of the ovalbumin is 0.8g to 1.5g, the mass range of the myosin is 0.8g to 1.5g, the mass of the platelet lysate is at least 7.5 times of the mass of the myosin, and the balance is the buffer solution. The invention has the beneficial effects that: the invention solves the problems of toxic and side effect of the components of the traditional cell freezing solution and low cell freezing survival rate, and improves the survival rate of cells during freezing by changing the balance of electrolytes inside and outside the cells, enhancing the tension of cell membranes and armed cell membrane structure.
Description
Technical Field
The invention relates to the technical field of hematopoietic stem cell cryopreservation, in particular to a hematopoietic stem cell cryopreservation liquid and a preparation method and a using method thereof.
Background
The freezing solution is a substance capable of protecting cells from freezing damage, the cells can be protected from solution damage and ice crystal damage by adding the freezing solution into the cell suspension, and the concentration of electrolyte in unfrozen solution inside and outside the cells can be reduced by maintaining a certain molar concentration inside and outside the cells, so that the cells are protected from damage of solute.
When the traditional cell freezing solution is stored in a refrigerator at the temperature of minus 80 ℃, low molecular or high molecular chemicals are added, and the cell survival rate of the cells during low-temperature storage is stabilized by changing the balance of electrolytes inside and outside the cells and osmotic pressure; for example, the conventional cell freezing solution is added with 10% of dimethyl sulfoxide, albumin and hydroxyethyl starch composition, and some low molecular substances such as dimethyl sulfoxide have cytotoxicity and other toxic and side effects, the survival rate of frozen cells is not very high, and the conventional cell freezing protective solution has many defects in cell freezing on the whole.
Disclosure of Invention
The invention aims to solve the technical problem of the prior art and provides a hematopoietic stem cell frozen stock solution, a preparation method and a use method thereof.
The technical scheme for solving the technical problems is as follows:
according to one aspect of the present invention, there is provided a hematopoietic stem cell cryopreserved comprising lecithin, ovalbumin, myosin, platelet lysate and buffer;
wherein, in every 100ml of the frozen stock solution, the mass range of the lecithin is 1g to 4g, the mass range of the ovalbumin is 0.8g to 1.5g, the mass range of the myosin is 0.8g to 1.5g, the mass of the platelet lysate is at least 7.5 times of the mass of the myosin, and the balance is the buffer solution.
The invention has the beneficial effects that: the heavy chain of myosin can transport the ovalbumin into the cell, so that the ovalbumin is combined with water molecules in advance to change the viscosity in the cell, thereby preventing the formation of ice crystals when cooling, in addition, the combined action of myosin and lecithin can enhance the tension and the compressive resistance of cell membranes, thereby ensuring that the cell is not easy to be punctured by the ice crystals, thereby ensuring that the cell can be directly frozen and stored in a refrigerator at the temperature of-80 ℃ without program control cooling when being frozen and being convenient to operate; the platelet lysate can provide required ATP and nutrient substances for the transportation of myosin, and also can be used for regulating the electrolyte balance together with a buffer solution; the invention solves the problems of toxic and side effect of the components of the traditional cell freezing solution and low cell freezing survival rate, and improves the survival rate of cells during freezing by changing the balance of electrolytes inside and outside the cells, enhancing the tension of cell membranes and armed cell membrane structure.
On the basis of the technical scheme, the invention can be further improved as follows.
Further: the mass ratio of the lecithin to the myosin ranges from 4:1 to 1:1, and the mass ratio of the ovalbumin to the myosin ranges from 8:10 to 11:10, per 100ml of the frozen stock solution.
Further: the mass ratio of the lecithin to the myosin was 3:1 per 100ml of the frozen stock solution.
The beneficial effects of the further scheme are as follows: the efficiency of the cooperation of lecithin and myosin is improved, the formation of ice crystals is reduced as far as possible, and the cell cryopreservation does not need program control cooling and is convenient to operate.
Further: the mass ratio of the ovalbumin to the myosin is 1:1 per 100ml of the frozen stock solution.
The beneficial effects of the further scheme are as follows: the combined action efficiency of the ovalbumin and the myosin is improved, the tension and the compressive resistance of the cell membrane are enhanced as much as possible, the cells are prevented from being punctured by ice crystals, and the survival rate of the cells is improved.
Further: the buffer solution is Bomaili A injection or PBS buffer solution.
The beneficial effects of the further scheme are as follows: the functions of constant volume and electrolyte balance adjustment are achieved.
According to another aspect of the present invention, there is provided a method for preparing a frozen stock solution of hematopoietic stem cells, each 100ml of the frozen stock solution being prepared, comprising the steps of:
s01: uniformly mixing 1g to 4g of lecithin, 0.8g to 1.5g of ovalbumin and 0.8g to 1.5g of myosin in 2ml of Bomai A injection buffer solution to obtain a mixed solution;
s02: standing the mixed solution obtained in the step S01 for 15min at normal temperature in the dark;
s03: adding platelet lysate with the mass at least 7.5 times that of the myosin into the mixed solution, and adding a certain amount of Bomai force A injection buffer solution for constant volume to ensure that the volume of the frozen stock solution reaches 100 ml.
The invention has the beneficial effects that: lecithin, ovalbumin and myosin are evenly mixed in 2ml of Bomaili A injection buffer solution and are kept stand in the dark at normal temperature, so that biological action time is provided for the mutual combination of the lecithin, the myosin and the ovalbumin, and the cooperation efficiency of the lecithin, the myosin and the ovalbumin is improved.
On the basis of the technical scheme, the invention can be further improved as follows.
Further: in the step S01, the mass ratio of the lecithin to the myosin is 3: 1.
The beneficial effects of the further scheme are as follows: the efficiency of the cooperation of lecithin and myosin is improved, the formation of ice crystals is reduced as far as possible, and the cell cryopreservation does not need program control cooling and is convenient to operate.
Further: in the step S01, the mass ratio of the ovalbumin to the myosin is 1: 1.
The beneficial effects of the further scheme are as follows: the combined action efficiency of the ovalbumin and the myosin is improved, the tension and the compressive resistance of the cell membrane are enhanced as much as possible, the cells are prevented from being punctured by ice crystals, and the survival rate of the cells is improved.
According to another aspect of the present invention, there is provided a method for using a hematopoietic stem cell frozen stock solution, comprising the steps of adding the prepared hematopoietic stem cell suspension to the frozen stock solution, and directly storing in a refrigerator at-80 ℃.
The invention has the beneficial effects that: when the cells are frozen, the cells do not need to be cooled by program control, can be directly frozen in a refrigerator with the temperature of minus 80 ℃, and are convenient to operate.
On the basis of the technical scheme, the invention can be further improved as follows.
Further: the volume ratio of the freezing medium to the hematopoietic stem cell suspension is 1:1 or 2: 1.
The beneficial effects of the further scheme are as follows: the cells can fully contact and interact with the active ingredients of the cryopreservation liquid, so that the optimal cryopreservation effect is achieved, and the cell survival rate is improved.
Detailed Description
The principles and features of this invention are described below in conjunction with examples which are set forth to illustrate, but are not to be construed to limit the scope of the invention.
A hematopoietic stem cell cryopreserved comprising lecithin, ovalbumin, myosin, platelet lysate, and a buffer;
wherein, in every 100ml of the frozen stock solution, the mass range of the lecithin is 1g to 4g, the mass range of the ovalbumin is 0.8g to 1.5g, the mass range of the myosin is 0.8g to 1.5g, the mass of the platelet lysate is at least 7.5 times of the mass of the myosin, and the balance is the buffer solution.
The mass ratio of the lecithin to the myosin ranges from 4:1 to 1:1, and the mass ratio of the ovalbumin to the myosin ranges from 8:10 to 11:10, per 100ml of the frozen stock solution.
Preferably, the mass ratio of lecithin to myosin is 3:1 per 100ml of the frozen stock solution.
Preferably, the mass ratio of the ovalbumin to the myosin is 1:1 per 100ml of the frozen stock solution.
The buffer solution is Bo Mai Li A injection solution or PBS buffer solution, and in the embodiment, Bo Mai Li A injection buffer solution is preferred.
The Bomaili A injection buffer was purchased from Shanghai Baite medical supplies, Inc.
Preferably, the frozen stock solution contains 3g of lecithin, 1g of ovalbumin, 1g of myosin and 7.5g of platelet lysate per 100ml, and the balance is Bo Mai injection buffer.
A method for preparing a hematopoietic stem cell frozen stock solution, wherein each 100ml of the frozen stock solution is prepared, and the method comprises the following steps:
s01: uniformly mixing 1g to 4g of lecithin, 0.8g to 1.5g of ovalbumin and 0.8g to 1.5g of myosin in 2ml of Bomai A injection buffer solution to obtain a mixed solution;
s02: standing the mixed solution obtained in the step S01 for 15min at normal temperature in the dark;
s03: adding platelet lysate with the mass at least 7.5 times that of the myosin into the mixed solution, and adding a certain amount of Bomai force A injection buffer solution for constant volume to ensure that the volume of the frozen stock solution reaches 100 ml.
In step S01, the mass ratio of lecithin to myosin is 3:1, and the mass ratio of ovalbumin to myosin is 1: 1.
A method for using the frozen stock solution of hematopoietic stem cells includes such steps as adding the prepared hematopoietic stem cell suspension to said frozen stock solution, and directly storing in-80 deg.C refrigerator.
Further, the volume ratio of the freezing medium to the hematopoietic stem cell suspension is 1:1 or 2: 1. In this embodiment, the volume ratio of the cryopreservation solution to the hematopoietic stem cell suspension is preferably 2: 1.
The first embodiment is as follows: a hematopoietic stem cell frozen stock solution is prepared by 100ml according to a preparation method, wherein the hematopoietic stem cell frozen stock solution comprises 3g of lecithin, 1g of ovalbumin, 1g of myosin and 7.5g of platelet lysate, and the balance is Bo Mai injection buffer solution, 100ml of hematopoietic stem cell suspension is added into the frozen stock solution according to a using method, and then the frozen stock solution is placed into a refrigerator at the temperature of 80 ℃ below zero for storage.
Example two: a hematopoietic stem cell frozen stock solution is prepared by 100ml according to a preparation method, wherein the hematopoietic stem cell frozen stock solution comprises 3g of lecithin, 1g of ovalbumin, 1g of myosin and 7.5g of platelet lysate, and the balance is Bo Mai injection buffer solution, 50ml of hematopoietic stem cell suspension is added into the frozen stock solution according to a using method, and then the frozen stock solution is placed into a refrigerator at the temperature of 80 ℃ below zero for storage.
Example three: a hematopoietic stem cell frozen stock solution is prepared by 100ml according to a preparation method, wherein the hematopoietic stem cell frozen stock solution comprises 1g of lecithin, 1g of ovalbumin, 1g of myosin, 7.5g of platelet lysate and the balance of Bo Mai injection buffer solution, 50ml of hematopoietic stem cell suspension is added into the frozen stock solution according to a using method, and then the frozen stock solution is placed into a refrigerator at the temperature of 80 ℃ below zero for storage.
Example four: a hematopoietic stem cell frozen stock solution is prepared by 100ml according to a preparation method, wherein the frozen stock solution comprises 3g of lecithin, 0.8g of ovalbumin, 1g of myosin and 7.5g of platelet lysate, and the balance is Bo Mai injection buffer solution, 50ml of hematopoietic stem cell suspension is added into the frozen stock solution according to a using method, and then the frozen stock solution is placed into a refrigerator at the temperature of 80 ℃ below zero for storage.
Example five: a hematopoietic stem cell frozen stock solution is prepared by 100ml according to a preparation method, wherein the hematopoietic stem cell frozen stock solution comprises 3g of lecithin, 1g of ovalbumin, 1g of myosin and 6g of platelet lysate, and the balance is Bomaili A injection buffer solution, 50ml of hematopoietic stem cell suspension is added into the frozen stock solution according to a using method, and then the frozen stock solution is placed into a refrigerator at the temperature of 80 ℃ below zero for storage.
The results of the experimental data are shown in table 1, table 2 and table 3 below, respectively, after the hematopoietic stem cells of the five groups of examples were recovered before and after one month of cryopreservation of the hematopoietic stem cells, and the recovery rate, survival rate, CD34+ recovery rate and colony forming ability of the hematopoietic stem cells were measured.
TABLE 1 hematopoietic stem cell survival data Table
Among them, TNC means nucleated cells, hematopoietic stem cells are contained in the nucleated cells, and the recovery rate of hematopoietic stem cells is counted by counting the number of nucleated cells, and the survival rate of hematopoietic stem cells is counted by trypan blue staining.
TABLE 2 data sheet for CD34+ recovery of cells
TABLE 3 data sheet of colony Forming ability of cells
CFU (Colony-Forming Units), which is the total number of colonies of stem cells per unit volume, is used to express the proliferation ability of stem cells.
As is clear from the experimental result data in tables 1, 2 and 3, the survival rate of cells after cryopreservation of hematopoietic stem cells, the recovery rate of CD34+ of cells and the proliferation potency of cells were all the best under the experimental conditions in example two; reducing the amount of lecithin, ovalbumin, or platelet lysate can affect the cryopreservation effect of the cells.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A hematopoietic stem cell cryopreservation liquid is characterized in that: including lecithin, ovalbumin, myosin, platelet lysate and buffers;
wherein, in every 100ml of the frozen stock solution, the mass range of the lecithin is 1g to 4g, the mass range of the ovalbumin is 0.8g to 1.5g, the mass range of the myosin is 0.8g to 1.5g, the mass of the platelet lysate is at least 7.5 times of the mass of the myosin, and the balance is the buffer solution.
2. The hematopoietic stem cell cryopreserved fluid according to claim 1, wherein: the mass ratio of the lecithin to the myosin ranges from 4:1 to 1:1, and the mass ratio of the ovalbumin to the myosin ranges from 8:10 to 11:10, per 100ml of the frozen stock solution.
3. The hematopoietic stem cell cryopreserved fluid according to claim 2, wherein: the mass ratio of the lecithin to the myosin was 3:1 per 100ml of the frozen stock solution.
4. The hematopoietic stem cell cryopreserved fluid according to claim 2, wherein: the mass ratio of the ovalbumin to the myosin is 1:1 per 100ml of the frozen stock solution.
5. The hematopoietic stem cell cryopreserved fluid according to claim 1, wherein: the buffer solution is Bomaili A injection or PBS buffer solution.
6. A method for preparing a hematopoietic stem cell frozen stock solution, wherein each 100ml of the frozen stock solution is prepared, comprising the steps of:
s01: uniformly mixing 1g to 4g of lecithin, 0.8g to 1.5g of ovalbumin and 0.8g to 1.5g of myosin in 2ml of Bomai A injection buffer solution to obtain a mixed solution;
s02: standing the mixed solution obtained in the step S01 for 15min at normal temperature in the dark;
s03: adding platelet lysate with the mass at least 7.5 times that of the myosin into the mixed solution, and adding a certain amount of Bomai force A injection buffer solution for constant volume to ensure that the volume of the frozen stock solution reaches 100 ml.
7. The method according to claim 6, wherein the method comprises the steps of: in the step S01, the mass ratio of the lecithin to the myosin is 3: 1.
8. The method according to claim 6, wherein the method comprises the steps of: in the step S01, the mass ratio of the ovalbumin to the myosin is 1: 1.
9. A method for using a hematopoietic stem cell frozen stock solution is characterized by comprising the following steps: the method comprises the following steps of adding the prepared hematopoietic stem cell suspension into the frozen stock solution, and directly storing in a refrigerator at-80 ℃.
10. The method of using a hematopoietic stem cell cryopreserved fluid according to claim 9, wherein the method comprises the steps of: the volume ratio of the freezing medium to the hematopoietic stem cell suspension is 1:1 or 2: 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810901210.6A CN110810397A (en) | 2018-08-09 | 2018-08-09 | Hematopoietic stem cell cryopreservation liquid and preparation method and application method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810901210.6A CN110810397A (en) | 2018-08-09 | 2018-08-09 | Hematopoietic stem cell cryopreservation liquid and preparation method and application method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110810397A true CN110810397A (en) | 2020-02-21 |
Family
ID=69541332
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810901210.6A Pending CN110810397A (en) | 2018-08-09 | 2018-08-09 | Hematopoietic stem cell cryopreservation liquid and preparation method and application method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110810397A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102349500A (en) * | 2011-11-10 | 2012-02-15 | 成都清科生物科技有限公司 | Mesenchymal stem cell self-preserving liquid |
| US20150037783A1 (en) * | 2012-03-14 | 2015-02-05 | Membrane Protective Technologies, Inc. | System and Substances for Cryopreservation of Viable Cells |
| CN107027743A (en) * | 2017-06-14 | 2017-08-11 | 深圳市泰华细胞工程有限公司 | Cells frozen storing liquid and cell freezing method |
| CN107156111A (en) * | 2017-06-30 | 2017-09-15 | 陈印平 | A kind of candidate stem cell cell cryopreservation agent |
| CN108094404A (en) * | 2017-12-20 | 2018-06-01 | 北京臻惠康生物科技有限公司 | A kind of improved mesenchyme stem cell protection solution and application thereof |
| CN108617638A (en) * | 2017-03-22 | 2018-10-09 | 拜西欧斯（北京）生物技术有限公司 | Tissue and/or cell cryopreservation protection liquid and its preparation and application |
-
2018
- 2018-08-09 CN CN201810901210.6A patent/CN110810397A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102349500A (en) * | 2011-11-10 | 2012-02-15 | 成都清科生物科技有限公司 | Mesenchymal stem cell self-preserving liquid |
| US20150037783A1 (en) * | 2012-03-14 | 2015-02-05 | Membrane Protective Technologies, Inc. | System and Substances for Cryopreservation of Viable Cells |
| CN108617638A (en) * | 2017-03-22 | 2018-10-09 | 拜西欧斯（北京）生物技术有限公司 | Tissue and/or cell cryopreservation protection liquid and its preparation and application |
| CN107027743A (en) * | 2017-06-14 | 2017-08-11 | 深圳市泰华细胞工程有限公司 | Cells frozen storing liquid and cell freezing method |
| CN107156111A (en) * | 2017-06-30 | 2017-09-15 | 陈印平 | A kind of candidate stem cell cell cryopreservation agent |
| CN108094404A (en) * | 2017-12-20 | 2018-06-01 | 北京臻惠康生物科技有限公司 | A kind of improved mesenchyme stem cell protection solution and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| LEBKOWSKI J.S.等: "Serum‐free culture of hematopoietic stem cells: A Review", 《STEM CELLS JOURNALS》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chaytor et al. | Inhibiting ice recrystallization and optimization of cell viability after cryopreservation | |
| CN107027743B (en) | Cell cryopreservation solution and cell cryopreservation method | |
| Koshimoto et al. | Effect of osmolality and oxygen tension on the survival of mouse sperm frozen to various temperatures in various concentrations of glycerol and raffinose | |
| Fuller et al. | Biopreservation of hepatocytes: current concepts on hypothermic preservation, cryopreservation, and vitrification | |
| CN104839144A (en) | Vitrification freezing fluid of oocytes | |
| CN108990964B (en) | Cell cryopreservation liquid | |
| CN107996558A (en) | Cells frozen storing liquid and its application | |
| CN102771472B (en) | Freezing liquid for preserving embryo, preparation method and application thereof | |
| Wowk | How cryoprotectants work | |
| CN113490414B (en) | Preservation of Stem cells | |
| Zhang et al. | Effects of four disaccharides on nucleation and growth of ice crystals in concentrated glycerol aqueous solution | |
| Yang et al. | A hemocompatible cryoprotectant inspired by freezing-tolerant plants | |
| CN110810397A (en) | Hematopoietic stem cell cryopreservation liquid and preparation method and application method thereof | |
| Zhang et al. | Ion-specific effects on the growth of single ice crystals | |
| Uchida et al. | Trehalose uptake and dehydration effects on the cryoprotection of CHO–K1 cells expressing TRET1 | |
| CN111357739A (en) | Vitrified refrigerating fluid and production method thereof | |
| CN114190368B (en) | Serum-free immune cell cryopreservation liquid, preparation method and immune cell cryopreservation method | |
| CN115152740A (en) | Long-term pig tissue preservation solution and using method thereof | |
| JP6626225B2 (en) | Cell cryopreservation composition and cryopreservation method | |
| US10611996B2 (en) | Preservation and storage of biological specimens | |
| Nei | Mechanism of Haemolysis of Erythrocytes by Freezing, with Special Reference to Freezing at Near‐Zero Temperatures | |
| EP2611290A2 (en) | Method for cryopreservation of human spermatozoa free from seminal plasma using a fast and simple aseptic vitrification-devitrification process; portable kit for carrying out the method; and use of the same for treatment of disorders related to reproductive failures | |
| US20220354850A1 (en) | Anti-ischemic compositions | |
| Zhuang et al. | Cryopreservation of filaments of Scytosiphon lomentaria by vitrification | |
| CN106922651A (en) | A kind of cells frozen storing liquid of high activity |
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 |
Application publication date: 20200221 |
|
| RJ01 | Rejection of invention patent application after publication |