CN108739798A - A kind of cells frozen storing liquid - Google Patents
A kind of cells frozen storing liquid Download PDFInfo
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- CN108739798A CN108739798A CN201810979369.XA CN201810979369A CN108739798A CN 108739798 A CN108739798 A CN 108739798A CN 201810979369 A CN201810979369 A CN 201810979369A CN 108739798 A CN108739798 A CN 108739798A
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- Prior art keywords
- cell
- storing liquid
- stock solution
- frozen
- trehalose
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Classifications
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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
- 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
Abstract
The present invention provides a kind of cells frozen storing liquid without dimethyl sulfoxide (DMSO) and animal derived components, the frozen stock solution is made of trehalose, hydroxyethyl starch 130, sodium chloride, human serum albumin and distilled water.The advantages that frozen stock solution has high anabiosis rate, and ingredient is simple, and stability is good.
Description
Technical field
The present invention relates to technical field of cell biology, and in particular to a kind of cells frozen storing liquid.
Background technology
Stem cell and immunocyte technology extensive use at present, cell storage are the important rings in entire cell industrial chain
Section.Cell cryopreservation needs, using protective agent, to prevent cell from freezing and being damaged by ice crystal damage, dehydration and water suction in resuscitation process
Wound.Most widely used protective agent is dimethyl sulfoxide (DMSO) (DMSO), it is a kind of permeability protective agent, can reduce cell freezing point,
The formation of ice crystal is reduced, mitigates free radical to cell damage, changes biomembrane to electrolyte, drug, poisonous substance and metabolite
Permeability.But it studies have shown that there are certain toxic effects by DMSO, has an effect with protein hydrophobic group, leads to protein
Denaturation has vascular toxicity and liver renal toxicity, and there are potential carcinogenic risks.DMSO can be very good freezing and resuscitation process
Middle protection cell, but the cell after recovering is cultivated, and DMSO could be discharged by the metabolizing cells of certain time,
Cell can just carry out next step application at this time.Because of this process so that stem cell receives considerable restraint in the application.
It generally to use fetal calf serum or calf serum as protective agent in cells frozen storing liquid, protect cell membrane, improve and freeze
Deposit the cell survival rate of resuscitation process.But crazy heifer disease virus may be contained in cow's serum, endotoxin, exotoxin is also potential
Unknown virus risk.
Trehalose is by two glucose molecules with a, a, 1, and 1- glycosidic bonds constitute nonreducing sugar, and self property is very steady
It is fixed, protective film can be formed in cell surface, be effectively protected that protein molecule is indeformable, and the life process for the body that sustains life
And biological characteristic.
Invention content
The purpose of the present invention is to provide a kind of cells frozen storing liquid, the frozen stock solution does not add DMSO and animal derived components.
To achieve the goals above, present invention employs following technical solutions：
A kind of cells frozen storing liquid, the frozen stock solution include the component of following w/v：2-15% trehaloses, 1-8%
The distilled water of hydroxyethyl starch 130,0.1-1.5% sodium chloride, 0.5-1.0% human serum albumins and surplus.
Preferably, a kind of cells frozen storing liquid, the frozen stock solution include the component of following w/v：The seaweed of 4-12%
Sugar, the hydroxyethyl starch 130 of 2-6%, the sodium chloride of 0.3-1.1%, the human serum albumin of 1-8% and the distilled water of surplus.
Preferably, a kind of cells frozen storing liquid, the frozen stock solution include the component of following w/v：5-8% trehaloses,
2-5% hydroxyethyl starch 130,0.4-1.0% sodium chloride, 3-6% human serum albumins and surplus distilled water.
Preferably, a kind of cells frozen storing liquid, the frozen stock solution include the component of following w/v：5% trehalose,
2% 130,0.9% sodium chloride of hydroxyethyl starch, the distilled water of 3% human serum albumin and surplus.
Preferably, a kind of cells frozen storing liquid, the frozen stock solution include the component of following w/v：6% trehalose,
3% 130,0.9% sodium chloride of hydroxyethyl starch, the distilled water of 4% human serum albumin and surplus.
Preferably, a kind of cells frozen storing liquid, the frozen stock solution include the component of following w/v：7% trehalose,
4% 130,0.9% sodium chloride of hydroxyethyl starch, the distilled water of 6% human serum albumin and surplus.
Preferably, a kind of cells frozen storing liquid, the frozen stock solution include the component of following w/v：8% trehalose,
5% 130,0.9% sodium chloride of hydroxyethyl starch, the distilled water of 5% human serum albumin and surplus.
A kind of preparation method of cells frozen storing liquid, includes the following steps：
1) trehalose is weighed, is dissolved in distilled water；
2) addition hydroxyethyl starch 130, sodium chloride and human serum albumin into the step 1)；
3) by mixed liquor biofilter degerming, cells frozen storing liquid is obtained.
The present invention has the following advantages：
The cells frozen storing liquid of the present invention does not contain DMSO and animal derived components, using bio-safety ingredient, and frozen stock solution at
Divide simply, it is at low cost；The high recovery motility rate of cell is kept simultaneously, does not influence the characteristic of cell and the proliferative capacity of cell.
Description of the drawings：
Fig. 1 is to use recovery stem cell adhere-wall culture 4X photomicrographs in 24 hours after No. 1 frozen stock solution in embodiment 2.
Fig. 2 is to use recovery stem cell adhere-wall culture 4X photomicrographs in 24 hours after No. 2 frozen stock solutions in embodiment 2.
Fig. 3 is to use recovery stem cell adhere-wall culture 4X photomicrographs in 24 hours after No. 3 frozen stock solutions in embodiment 2.
Fig. 4 is to use recovery stem cell adhere-wall culture 4X photomicrographs in 24 hours after No. 4 frozen stock solutions in embodiment 2.
Specific implementation mode
Invention is described in further detail with specific implementation mode below.
The present invention is described with reference to specific embodiment, but it is clear that still various modifications may be made and transformation
And without prejudice to the spirit and scope of the present invention.Therefore, the description and the appended drawings of the invention are it is believed that be illustrative rather than limit
Property processed.
Specific embodiment：
Embodiment 1.
A kind of cells frozen storing liquid, including：Trehalose, hydroxyethyl starch 130, sodium chloride, human serum albumin and distilled water.
Frozen stock solution 1, content of trehalose 5%, 130 content 2% of hydroxyethyl starch, sodium chloride 0.9%, human serum albumin 3%
And distilled water.
Frozen stock solution 2, content of trehalose 6%, 130 content 3% of hydroxyethyl starch, sodium chloride 0.9%, human serum albumin 4%
And distilled water.
Frozen stock solution 3, content of trehalose 7%, 130 content 4% of hydroxyethyl starch, sodium chloride 0.9%, human serum albumin 6%
And distilled water.
Frozen stock solution 4, content of trehalose 8%, 130 content 5% of hydroxyethyl starch, sodium chloride 0.9%, human serum albumin 5%
And distilled water.
It is configured in accordance with the following steps：
1) trehalose is weighed, is dissolved in distilled water 10mL；
2) addition hydroxyethyl starch 130, sodium chloride and human serum albumin into the step 1)；
3) by mixed liquor biofilter degerming, cells frozen storing liquid is obtained.
The biofilter is 0.22 μm of bacterial filter.
Embodiment 2.
It is tested using human umbilical cord mesenchymal stem cells, the cells frozen storing liquid configured to embodiment 1 is tested.Cell
Culture and cell cryopreservation use following steps：
1) micro- sem observation cell growth to density about 95% when frozen；
2) culture solution is sucked out, 15mL physiological saline is added, gently shakes, abandons physiological saline；
3) pancreatin 3mL is added in culture bottle, and room temperature digests 5min, and micro- sem observation cell all suspends, and α-MEM trainings are added
Base (containing serum substitute 10%) 5mL is supported, digestion is terminated；
4) it draws cell suspension and is put into centrifuge tube, 1100rpm centrifuges 5min；
5) after centrifuging, supernatant is abandoned, physiological saline 45mL is added, draws a drop cell count 9 × 106A cell, cell are lived
Rate 97%, 1100rpm centrifuge 5min；
6) physiological saline is abandoned, No. 1-4 4 DEG C of precoolings of frozen stock solution of embodiment 1, adjustment cell concentration 3-9 × 10 are added6It is a thin
Born of the same parents/mL frozen stock solutions, every cryopreservation tube refinement cytosol about 1.5mL；
7) cryopreservation tube is put into programmed cooling instrument (Thermo Scientific Forma CryoMed Controlled
Rate Freezer 7451), start ready-made program, completion in about 1.5 hours freezes；
8) directly cell is transferred in liquid nitrogen and is frozen for a long time.
Cell recovery
Cell is taken out from liquid nitrogen, is put into rapidly in 37-40 DEG C of water-bath, notices that cell liquid level has to be completely immersed in
The water surface is higher than the water surface hereinafter, freezing nozzle, and quickly rocking makes cell melt in 1min, waits for the ice for having grain of rice size in liquid
Crystalline substance can take out from water-bath.Cell after recovery is added directly into corresponding culture medium and is cultivated.Second after recovery
Its micro- sem observation cell, takes pictures.It is as shown in Figures 1 to 4 using frozen stock solution 1-4 frozen stock solution photos.It is thin with the observation of 4X object lens
Born of the same parents, as shown in the photo, the adherent ratio of cell>90%, only a small amount of non-attached cell, attached cell fully stretch in each visual field
Exhibition, form is uniform, cell no abnormality seen form.
Cell count after recovery：
From the cell suspension after recovery, 20 microlitres are drawn, 20 microlitres of the 0.08% trypan blue working solution configured is added,
Gently mixing prevents bubble from generating, and inhales 20 microlitres, is added in special numeration plate, is carried out using Countstar cell counters
Cell quantity and survival rate test, are repeated twice.Experimental result is as shown in table 1.
Table 1 the result shows that, freeze before and recovery after, total number of cells, concentration, motility rate, diameter without occur significant changes,
Show the present invention frozen stock solution can during freezing effective protection stem cell.
Table 1. is using various concentration frozen stock solution the stem cell cell number and survival rate front and back in recovery
The step of cck8 cell Proliferation viability examinations, is as follows：
1) one bottle of umbilical cord mesenchymal stem cells in culture is taken, is carried out when micro- sem observation cell growth is to density about 95%
Digestion；
2) culture solution is sucked out, 15mL physiological saline is added, gently shakes, abandons physiological saline；
3) pancreatin 3mL, room temperature 5min is added in culture bottle, and micro- sem observation cell all suspends, and α-MEM culture mediums are added
(containing serum substitute 10%) 5mL, terminates digestion；
4) it draws cell suspension and is put into centrifuge tube, 1100rpm centrifuges 5min, uses brine 2 times；
5) after centrifuging, supernatant is abandoned, special culture media 1mL is added, a drop cell count is drawn, calculates adjustment cell concentration extremely
5×104A cell/100 microlitre (culture medium dilution) take 100 microlitres, repeatedly 3 samples, 96 porocyte culture plates of addition, 37 DEG C
5%CO2 cultures are supported cultivates 2 hours in case.
6) it recovers the cell that is frozen using 1-4 frozen stock solutions in embodiment 1 by above-mentioned recovery step, on cell suspension is direct
Cell counter counts, and cell concentration is adjusted to 5 × 10 with special culture media4A cell/100 microlitre (culture medium dilution), take
100 microlitres, 3 samples are repeated, 96 porocyte culture plates are added, 37 DEG C of 5%CO2 cultures are supported and cultivated 2 hours in case.
7) frozen stock solution containing DMSO (10%DMSO+90%FBS) freezing and storing umbilical mesenchymal stem cells are used, using same recovery
Method, the directly upper cell counter of cell suspension count, and cell concentration is adjusted to 5 × 10 with special culture media4A cell/100
Microlitre (culture medium dilution) takes 100 microlitres, repeats 3 samples, and 96 porocyte culture plates are added, and 37 DEG C of 5%CO2 cultures are supported in casees
Culture 2 hours.
7) in detection hole, 10 microlitres of CCK8 solution is added per hole, continues to be put into incubator and cultivate 2 hours.
8) culture plate is put into microplate reader, 450nm wavelength, detects light absorption value.
The results are shown in Table 2, compared with normally culture cell, is frozen using 1 to 4 kinds of frozen stock solutions of the embodiment of the present invention thin
Light absorption value of the born of the same parents after the detection of 450nm microplate reader is slightly lower, close with the light absorption value of the cell frozen using DMSO frozen stock solutions, shows
The frozen stock solution of the present invention can make cell keep preferable proliferative capacity.
Table 2. uses various concentration frozen stock solution stem cell cell viability before and after recovery
Note：It is 5*10 per hole cell4A, cell viability is higher, and the light absorption value of detection is higher.
Blank control：Blank well is not added with cell, only adds 100 microlitres of special culture medias, and 10 microlitres of 8 reagents of cck-are added.
Embodiment 3.
It is tested using human cord blood extraction candidate stem cell, candidate stem cell and mescenchymal stem cell belong to inhomogeneity
The cell of type, candidate stem cell are the progenitor cells of immunocyte, cell dia, cellular morphology, cell characteristics and the culture that suspends
Immunocyte is similar, and suspension cell is represented by taking candidate stem cell as an example, detects novel liquid storage.Embodiment 1 is configured No. 1-4 thin
Born of the same parents' frozen stock solution is tested.Cell detaches and cell cryopreservation uses following steps：
1) the qualified Cord blood of detection, takes 15mL and PBS (pH7.0) 15mL mixings
2) 15mL lymphocyte separation mediums are added in 50mL centrifuge tubes, are slowly added to the PBS mixed and Cord blood
Solution
3) it is put into centrifuge, 400g centrifuges 30min
4) tunica albuginea layer in the middle part of centrifuge tube is carefully drawn, PBS rinsings are added, centrifuge 2000rpm 10min, wash repeatedly 3 times
5) final umbilical cord blood hematopoietic stem cell is harvested, these cells, which are divided equally, is directly added into the example 1 of 4 DEG C of precoolings No. 1-4
In frozen stock solution, cell concentration 1-9 × 10 are adjusted7A cell/mL frozen stock solutions, every cryopreservation tube refinement cytosol about 1.5mL
6) cryopreservation tube is put into programmed cooling instrument (Thermo Scientific Forma CryoMed Controlled
Rate Freezer 7451), start ready-made program, completion in about 1.5 hours freezes；
7) directly cell is transferred in liquid nitrogen and is frozen for a long time.
Cell recovery
Cell is taken out from liquid nitrogen, is put into rapidly in 37-40 DEG C of water-bath, notices that cell liquid level has to be completely immersed in
The water surface is higher than the water surface hereinafter, freezing nozzle, and quickly rocking makes cell melt in 1min, waits for the ice for having grain of rice size in liquid
Crystalline substance can take out from water-bath, with reference to method for cell count in embodiment 2, draw 20 microlitres of cells and carry out numeration detection work
Rate (table 3).Cell is directly placed into immunocyte special culture media, carries out suspension culture.
Table 3 the result shows that, freeze before and recovery after, total number of cells, concentration, motility rate without occur significant changes, into one
Step show the present invention cells frozen storing liquid can during freezing effective protection immunocyte.
Table 3. is using various concentration frozen stock solution the stem cell cell number and survival rate front and back in recovery
Claims (8)
1. a kind of cells frozen storing liquid, the frozen stock solution includes the component of following w/v：Trehalose, the 1-8% of 2-15%
Hydroxyethyl starch 130, the sodium chloride of 0.1-1.5%, the human serum albumin of 0.5-10% and surplus distilled water.
2. cells frozen storing liquid according to claim 1, the frozen stock solution includes the component of following w/v：4-12%
Trehalose, the hydroxyethyl starch 130 of 2-6%, the sodium chloride of 0.3-1.1%, the human serum albumin of 1-8% and the distillation of surplus
Water.
3. cells frozen storing liquid according to claim 1, the frozen stock solution includes the component of following w/v：5-8%
Trehalose, 2-5% hydroxyethyl starch 130,0.4-1.0% sodium chloride, 3-6% human serum albumins and surplus distilled water.
4. cells frozen storing liquid according to claim 3, the frozen stock solution includes the component of following w/v：5%
The distilled water of trehalose, 2% 130,0.9% sodium chloride of hydroxyethyl starch, 3% human serum albumin and surplus.
5. cells frozen storing liquid according to claim 3, the frozen stock solution includes the component of following w/v：6%
The distilled water of trehalose, 3% 130,0.9% sodium chloride of hydroxyethyl starch, 4% human serum albumin and surplus.
6. cells frozen storing liquid according to claim 3, the frozen stock solution includes the component of following w/v：7%
The distilled water of trehalose, 4% 130,0.9% sodium chloride of hydroxyethyl starch, 6% human serum albumin and surplus.
7. cells frozen storing liquid according to claim 3, the frozen stock solution includes the component of following w/v：8%
The distilled water of trehalose, 5% 130,0.9% sodium chloride of hydroxyethyl starch, 5% human serum albumin and surplus.
8. a kind of preparation method of cells frozen storing liquid, includes the following steps：
1) trehalose is weighed, is dissolved in distilled water；
2) addition hydroxyethyl starch 130, sodium chloride and human serum albumin into the step 1)；
3) by mixed liquor biofilter degerming, cells frozen storing liquid is obtained.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109315386A (en) * | 2018-12-12 | 2019-02-12 | 中南大学湘雅三医院 | A kind of frozen stock solution and cryopreservation methods can be used for candidate stem cell or lymphocyte |
| CN109845726A (en) * | 2019-02-27 | 2019-06-07 | 李冲杰 | A kind of mescenchymal stem cell Cryopreservation protective agent |
| CN109907036A (en) * | 2019-02-27 | 2019-06-21 | 北京益华生物科技有限公司 | Cells frozen storing liquid, cell cryopreservation liquid and preparation method thereof and cell freezing method |
| CN112998009A (en) * | 2021-03-31 | 2021-06-22 | 北京益华生物科技有限公司 | NK cell cryopreservation liquid and preparation method and application thereof |
| CN113170779A (en) * | 2021-03-31 | 2021-07-27 | 北京益华生物科技有限公司 | Cell cryopreservation liquid, preparation method and application thereof, and cryopreservation method and feedback method of mononuclear cells |
| CN113973805A (en) * | 2021-10-25 | 2022-01-28 | 北京京蒙细胞生物科技股份有限公司 | Cell cryopreservation kit and using method thereof |
| CN114258910A (en) * | 2022-01-11 | 2022-04-01 | 华东理工大学 | Cell cryopreservation solution capable of being infused and application thereof |
| CN114762496A (en) * | 2021-01-11 | 2022-07-19 | 南京艾尔普再生医学科技有限公司 | Cardiac muscle cell freezing medium and its preparation method |
| CN114762495A (en) * | 2021-01-11 | 2022-07-19 | 南京艾尔普再生医学科技有限公司 | Myocardial cell cryopreservation method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1857312A (en) * | 2006-03-28 | 2006-11-08 | 中国人民解放军军事医学科学院野战输血研究所 | Pretreating liquid and freeze protecting liquid for cryopreservation of erythrocyte and their application |
| CN102511471A (en) * | 2011-12-15 | 2012-06-27 | 成都清科生物科技有限公司 | Mesenchymal stem cell frozen stock solution and preparation method thereof |
| CN104857022A (en) * | 2015-05-21 | 2015-08-26 | 北京青藤谷禧干细胞科技研究院有限公司 | MSC (mesenchymal stem cell) injection as well as preparation and application thereof |
| CN108029677A (en) * | 2017-12-26 | 2018-05-15 | 重庆斯德姆生物技术有限公司 | The frozen stock solution and cryopreservation methods of a kind of endothelial progenitor cells |
-
2018
- 2018-08-22 CN CN201810979369.XA patent/CN108739798A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1857312A (en) * | 2006-03-28 | 2006-11-08 | 中国人民解放军军事医学科学院野战输血研究所 | Pretreating liquid and freeze protecting liquid for cryopreservation of erythrocyte and their application |
| CN102511471A (en) * | 2011-12-15 | 2012-06-27 | 成都清科生物科技有限公司 | Mesenchymal stem cell frozen stock solution and preparation method thereof |
| CN104857022A (en) * | 2015-05-21 | 2015-08-26 | 北京青藤谷禧干细胞科技研究院有限公司 | MSC (mesenchymal stem cell) injection as well as preparation and application thereof |
| CN108029677A (en) * | 2017-12-26 | 2018-05-15 | 重庆斯德姆生物技术有限公司 | The frozen stock solution and cryopreservation methods of a kind of endothelial progenitor cells |
Non-Patent Citations (2)
| Title |
|---|
| 刘建福 等: "《细胞工程》", 30 June 2014, 华中科技大学出版社 * |
| 刘江东 等: "《细胞生物学实验教程》", 30 June 2005, 武汉大学出版社 * |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109315386A (en) * | 2018-12-12 | 2019-02-12 | 中南大学湘雅三医院 | A kind of frozen stock solution and cryopreservation methods can be used for candidate stem cell or lymphocyte |
| CN109845726A (en) * | 2019-02-27 | 2019-06-07 | 李冲杰 | A kind of mescenchymal stem cell Cryopreservation protective agent |
| CN109907036A (en) * | 2019-02-27 | 2019-06-21 | 北京益华生物科技有限公司 | Cells frozen storing liquid, cell cryopreservation liquid and preparation method thereof and cell freezing method |
| CN114762496A (en) * | 2021-01-11 | 2022-07-19 | 南京艾尔普再生医学科技有限公司 | Cardiac muscle cell freezing medium and its preparation method |
| CN114762495A (en) * | 2021-01-11 | 2022-07-19 | 南京艾尔普再生医学科技有限公司 | Myocardial cell cryopreservation method |
| CN114762496B (en) * | 2021-01-11 | 2023-11-07 | 南京艾尔普再生医学科技有限公司 | Myocardial cell frozen stock solution and preparation method thereof |
| CN114762495B (en) * | 2021-01-11 | 2023-11-07 | 南京艾尔普再生医学科技有限公司 | Myocardial cell cryopreservation method |
| CN112998009A (en) * | 2021-03-31 | 2021-06-22 | 北京益华生物科技有限公司 | NK cell cryopreservation liquid and preparation method and application thereof |
| CN113170779A (en) * | 2021-03-31 | 2021-07-27 | 北京益华生物科技有限公司 | Cell cryopreservation liquid, preparation method and application thereof, and cryopreservation method and feedback method of mononuclear cells |
| CN113973805A (en) * | 2021-10-25 | 2022-01-28 | 北京京蒙细胞生物科技股份有限公司 | Cell cryopreservation kit and using method thereof |
| CN114258910A (en) * | 2022-01-11 | 2022-04-01 | 华东理工大学 | Cell cryopreservation solution capable of being infused and application thereof |
| CN114258910B (en) * | 2022-01-11 | 2022-12-27 | 华东理工大学 | Cell cryopreservation solution capable of being infused and application thereof |
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Application publication date: 20181106 |