CN109845725A - Cell transport saves liquid and its application - Google Patents
Cell transport saves liquid and its application Download PDFInfo
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- CN109845725A CN109845725A CN201910071242.2A CN201910071242A CN109845725A CN 109845725 A CN109845725 A CN 109845725A CN 201910071242 A CN201910071242 A CN 201910071242A CN 109845725 A CN109845725 A CN 109845725A
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Abstract
It is transported the present invention provides a kind of cell and saves liquid and its application.Wherein, cell transport saves the component in liquid with densimeter, comprising: 0.01~2.5mg/ml of glucose, 0.05~10mg/ml of sodium chloride, 0.05~3mg/ml of potassium chloride, 0.05~1mg/ml of calcium chloride, 0.05~20mg/ml of sodium lactate, remaining group are divided into water.Through the invention, the cell for solving the problems, such as that cell transport preservation liquid in the related technology saves can not easily be applied to clinical application, it realizes to be transported by cell and saves the usage mode that the cell that liquid saves is similar to drug, so that facilitated application is in the technical effect of clinical application.
Description
Technical field
The present invention relates to life science and medicine technology field, transported in particular to a kind of cell save liquid and its
Using.
Background technique
It is used about Simple transportation of the cell (the especially cell of adhere-wall culture) under suspended state and short term stored
Cell transport save liquid, liquid etc. is generally saved by the cell activity of complete chemical component and is formed.Such as Publication No.
Added with pH buffer, more in the cell-preservation liquid used in the Chinese patent application of CN108770836A, CN109090104A
Kind of homeo-osmosis agent and a variety of oxygen free radical scavengers, energy substrate needed for also added cell maintenance metabolism, ingredient compared with
For complexity, cell viability maintains differ for 3-5 days.
Cell transport in the related technology saves liquid, and not only ingredient is complex, at high cost.Also, due in the related technology
Cell transport save in liquid and there is most of chemical substance that can not be directly entered in human body, therefore the transport of these cells is protected
Liquid storage acts on limited in terms of pushing the simplification of cell clinical treatment.
Summary of the invention
It is transported the present invention provides a kind of cell and saves liquid and its application, at least to solve cell transport in the related technology
Save the problem of cell that liquid saves can not easily be applied to clinical application.
Liquid is saved in a first aspect, transporting the embodiment of the invention provides a kind of cell, the cell transport saves in liquid
Component is with densimeter, comprising: 0.01~2.5mg/ml of glucose, 0.05~10mg/ml of sodium chloride, 0.05~3mg/ of potassium chloride
Ml, 0.05~1mg/ml of calcium chloride, 0.05~20mg/ml of sodium lactate, remaining group are divided into water.
Second aspect transports the application for saving liquid, packet the embodiment of the invention provides cell described in a kind of first aspect
It includes:
Cell and cell transport are saved liquid to mix, cell suspension is made；
By the cell suspension move into container in, and keep the volume ratio of the cell suspension and air be 1:(0.2~
30)；
The container is stored at a temperature of 2~15 DEG C.
The cell transport provided through the embodiment of the present invention saves liquid and its application, and cell transport saves the component in liquid
With densimeter, comprising: 0.01~2.5mg/ml of glucose, 0.05~10mg/ml of sodium chloride, 0.05~3mg/ml of potassium chloride, chlorine
Change 0.05~1mg/ml of calcium, 0.05~20mg/ml of sodium lactate, remaining group is divided into water, solves cell transport in the related technology
The cell that preservation liquid saves can not easily be applied to the problem of clinical application, realize and transport the thin of preservation liquid preservation by cell
Born of the same parents are similar to the usage mode of drug, so that facilitated application is in the technical effect of clinical application.
Detailed description of the invention
The drawings described herein are used to provide a further understanding of the present invention, constitutes part of this application, this hair
Bright illustrative embodiments and their description are used to explain the present invention, and are not constituted improper limitations of the present invention.In the accompanying drawings:
Fig. 1, Fig. 2 and Fig. 3 are to save the mesenchymal stem cell that liquid saves using the cell transport of the embodiment of the present invention
The schematic diagram of motility rate variation in seven days.
Specific embodiment
The feature and exemplary embodiment of various aspects of the invention is described more fully below, in order to make mesh of the invention
, technical solution and advantage be more clearly understood, with reference to the accompanying drawings and embodiments, the present invention is further retouched in detail
It states.It should be understood that described herein, the specific embodiments are only for explaining the present invention, is not intended to limit the present invention.For ability
For field technique personnel, the present invention can be implemented in the case where not needing some details in these details.It is right below
The description of embodiment is used for the purpose of better understanding the present invention to provide by showing example of the invention.
It should be noted that, in this document, the terms "include", "comprise" or its any other variant are intended to non-row
His property includes, so that the process, method, article or equipment for including a series of elements not only includes those elements, and
And further include other elements that are not explicitly listed, or further include for this process, method, article or equipment institute it is intrinsic
Element.In the absence of more restrictions, the element limited by sentence " including ... ", it is not excluded that including described want
There is also other identical elements in the process, method, article or equipment of element.
A kind of transport of cell is provided in the present embodiment saves liquid, cell transport saves the component in liquid with densimeter,
Including but not limited to: 0.01~2.5mg/ml of glucose, 0.05~10mg/ml of sodium chloride, 0.05~3mg/ml of potassium chloride, chlorination
0.05~1mg/ml of calcium, 0.05~20mg/ml of sodium lactate, remaining component are water in whole or in part.Preferably, cell transport is protected
Amino acid and vitamin can also be added in liquid storage.
Cell transport saves the clinically used sodium chloride being added in liquid, potassium chloride, calcium chloride, sodium lactate for remaining molten
Osmotic pressure, electrolyte and the acid-base balance of liquid.
When saving liquid preservation using above-mentioned cell transport, low temperature helps to reduce cell metabolism and extend cell cell
Holding time.Although there is still a need for certain nutriments to be metabolized however, cell is in low metabolism state.In order to maintain
The metabolism of cell, cell transport in the related art save in liquid, and the usual amino acid for selecting energy efficiency relatively low is made
For energy matter, and glucose is selected to improve energy efficiency as energy matter in embodiments of the present invention.And process pair
Cell transport saves the use verifying of liquid, and discovery uses glucose to be able to extend cell guarantor as the cell-preservation liquid of energy matter
Deposit the time.
Preferably, cell transport saves the combination that liquid includes but is not limited to one or more of following amino acid:
Essential amino acid: 0.1~20mg/ml of methionine, 0.05~30mg/ml of valine, 0.2~20mg/ of lysine
Ml, 0.05~40mg/ml of isoleucine, 0.1~20mg/ml of phenylalanine, 0.2~30mg/ml of leucine, tryptophan 0.05~
20mg/ml, 0.05~30mg/ml of threonine, and
Nonessential amino acid: 0.05~30mg/ml of glycine, 0.05~30mg/ml of alanine, serine 0.05~
30mg/ml, 0.05~20mg/ml of 0.05~20mg/ml of aspartic acid, glutamic acid and its amine, 0.05~30mg/ml of proline,
0.05~40mg/ml of arginine, 0.05~30mg/ml of histidine, 0.05~30mg/ml of tyrosine, 0.05~50mg/ of cystine
ml。
Preferably, cell transport saves the combination that liquid includes but is not limited to one or more of following vitamin: dimension
Raw 0.05~150IU/ml of element A, 0.05~25ug/ml of vitamin B2,0.05~500IU/ml of vitamin D, vitamin E
0.05~30ug/ml, 0.5~300ug/ml of vitamin C.
There is provided in the present embodiment cell transport save liquid in amino acid and vitamin be optional components, if choosing
With these amino acid or vitamin, and which amino acid or vitamin is selected to be added to the component that cell transport saves liquid,
It can be accepted or rejected for different cell types.
In the present embodiment also according to experimental result, has studied cell transport and save in liquid under the experiment condition of each component
Optimal concentration.
Preferably, cell transport saves the combination that liquid includes one or more of following amino acid: methionine
2.5mg/ml, valine 3mg/ml, lysine 5mg/ml, isoleucine 2mg/ml, phenylalanine-3,4-quinone .5mg/ml, leucine 2mg/
Ml, tryptophan 2mg/ml, threonine 4mg/ml and glycine 3mg/ml, alanine 3.5mg/ml, serine 1.5mg/ml,
Aspartic acid 2mg/ml, glutamic acid and its amine 5.5mg/ml, proline 2.2mg/ml, arginine 4.6mg/ml, histidine
2.5mg/ml, tyrosine 3.5mg/ml, cystine 8.5mg/ml.
Preferably, cell transport saves the combination that liquid includes one or more of following vitamin: vitamin
A1.25IU/ml, vitamin B2 2.5ug/ml, vitamin D 10IU/ml, vitamin E 7.5ug/ml, vitamin C 25ug/ml.
Preferably, it includes: glucose 0.2mg/ml, sodium chloride 3mg/ml, potassium chloride that the transport of above-mentioned cell, which saves liquid,
1mg/ml, calcium chloride 0.8mg/ml, sodium lactate 8mg/ml.
It saves liquid in view of the cell transport of the relevant technologies and generally uses pH buffer, a variety of homeo-osmosis agents and a variety of
Oxygen free radical scavenger, there are also the other compositions of in addition to this cell metabolism, substantially ingredient is all complex；And the present invention is real
The cell transport preservation liquid ingredient for applying example offer is more simple.In addition, the cell transport of the relevant technologies save liquid it is most of at
Dividing not can enter human body, and limited to drug direction development function to the clinical treatment of cell, especially adhere-wall culture is thin
Born of the same parents must not need professional when carrying out clinical treatment in future and carry out enzymic digestion in Cytology Lab and use centrifuge, super-clean bench
Etc. equipment, and these hinder cell as drug easily for clinical treatment significantly；And it is provided in an embodiment of the present invention
It is all the ingredient that can directly input human body that cell transport, which saves liquid ingredient, is similar to drug to convenient clinical application.
A kind of application of cell transport preservation liquid is additionally provided in the present embodiment, is included the following steps:
Step 1, cell and cell transport are saved liquid to mix, cell suspension is made；
Step 2, by cell suspension move into container in, and keep the volume ratio of cell suspension and air be 1:(0.2~
30)；
Wherein, the purpose of step 2 is the oxygen demand in order to guarantee cell under low metabolism.
Step 3, container is stored at a temperature of 2~15 DEG C.
Wherein, in normal temperature environment, intracellular various enzymes and other biological macromolecular reaction are metabolized vigorous, oxygen consumption height, point
Secretion may also be larger.The a large amount of oxygen radicals and lipid peroxide that these physiology courses can generate.They must quilt in time
It removes, environment pH and osmotic pressure is otherwise caused to change, cause cell swelling and death.And at low temperature, cell metabolism is lower, this
The available very big alleviation of a little problems.2~15 DEG C of temperature as cell preservation are selected in the present embodiment, under experimental condition
Optimum temperature be 12 DEG C, the temperature can be usually used in by day insulin preservation incubation chamber obtain.
The cell of the present embodiment can be used for clinical treatment and be related to cell collection, culture amplification, cell diversification processing, thin
Multiple processes such as born of the same parents' transplantation treatment.These processes generally comprise laboratory, hospital in enforcement place, therefore can be related to cell
It transports and uses between different location.
Preferably, cell density is 1 × 10 in cell suspension2~1 × 108/ml；In order to guarantee above-mentioned cell density,
The cell collected can also be counted in step 1.
Preferably, in step 2, after cell suspension being moved into container, the volume ratio of cell suspension and air is kept
Example is 1:5.Experiment shows the motility rate for using the volume ratio of cell suspension and air to help to maintain cell for 1:5.
Optionally, above-mentioned cell transport saves the application of liquid further include: in clinical application cell suspension, directly from container
Middle taking-up cell suspension；It is added in 100ml physiological saline, human body is inputted by intravenous injection or other injection systems.
Below by experiment and its result saves liquid to the cell transport of the embodiment of the present invention and its application is illustrated.
Fig. 1 is that Trypan Blue detection mesenchymal stem cell comes the 7th day Cell viability data under enzymic digestion,
Use whole components described in the embodiment of the present invention (including whole amino acid and vitamin) and with optium concentration and
Experiment condition such as temperature is 12 DEG C, suspension and AIR Proportional are experiment that 1:5 is carried out.In Fig. 1, (namely 0d) is thin at the beginning
Because the reason of enzymic digestion damages, Cell viability was in 96% or so, the 7th day (namely 7d) Cell viability during born of the same parents collect
It is gradually lowered to 91% or so.
The weight that various kinds of cell such as mescenchymal stem cell, fibroblast transport is saved using above-mentioned identical experiment condition
Multiple experiment, can also obtain similar experimental result, show that the cell saved is able to maintain that seven days or so vigor.
Fig. 2 is that Trypan Blue detection mesenchymal stem cell comes the 7th day Cell viability data under enzymic digestion,
Use essential component described in the embodiment of the present invention (not including amino acid and vitamin) and with optium concentration and experiment
Condition such as temperature is 12 DEG C, suspension and AIR Proportional are experiment that 1:5 is carried out.In Fig. 2, Cell viability is declined, and the 7th
It falls below 88% or so.
Fig. 3 is that Trypan Blue detection mesenchymal stem cell comes the 7th day Cell viability data under enzymic digestion,
Use essential component described in the embodiment of the present invention (not including amino acid and vitamin) and with non-optimal concentration (grape
Sugared 0.05mg/ml, sodium chloride 2mg/ml, potassium chloride 2mg/ml, calcium chloride 0.6mg/ml, sodium lactate 6mg/ml) and non-optimal reality
Testing condition, (storage temperature selects 8 DEG C, the experiment that carries out for 1:2) of suspension and AIR Proportional.In Fig. 3, Cell viability under
Drop, falls below 82% or so on the 7th day.
In conclusion
It saves liquid in view of the cell transport of the relevant technologies and generally uses pH buffer, a variety of homeo-osmosis agents and a variety of
Oxygen free radical scavenger, there are also the other compositions of in addition to this cell metabolism, substantially ingredient is all complex；And the present invention is real
The cell transport preservation liquid ingredient for applying example offer is more simple.
In addition, most of ingredient that the cell transport of the relevant technologies saves liquid not can enter human body, to the clinic of cell
Treat it is limited to drug direction development function, especially the cell of adhere-wall culture future carry out clinical treatment when must not be not required to
Professional is wanted to carry out enzymic digestion in Cytology Lab and use the equipment such as centrifuge, super-clean bench, and these hinder cell picture significantly
Drug is the same to be easily used for clinical treatment；And cell provided in an embodiment of the present invention transport save liquid ingredient be all can be direct
The ingredient of human body is inputted, is similar to drug to convenient clinical application.
Cell transport preservation liquid the maintaining 3-5 days to cell viability of the relevant technologies, and it was proved that, using this
The cell transport that inventive embodiments provide saves liquid and maintains to transport up to 7 days or so with the cell of the relevant technologies to the vigor of cell
Defeated preservation liquid phase is than having certain advantage at aspect of holding time.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.All within the spirits and principles of the present invention, made any to repair
Change, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of cell transport saves liquid, which is characterized in that the cell transport saves the component in liquid with densimeter, comprising:
0.01~2.5mg/ml of glucose, 0.05~10mg/ml of sodium chloride, 0.05~3mg/ml of potassium chloride, 0.05~1mg/ of calcium chloride
Ml, 0.05~20mg/ml of sodium lactate, remaining group are divided into water.
2. cell transport according to claim 1 saves liquid, which is characterized in that the cell transport saves liquid further include:
Amino acid and vitamin.
3. cell transport according to claim 2 saves liquid, which is characterized in that it includes following that the cell transport, which saves liquid,
The combination of one or more of amino acid: 0.1~20mg/ml of methionine, 0.05~30mg/ml of valine, lysine
0.2~20mg/ml, 0.05~40mg/ml of isoleucine, 0.1~20mg/ml of phenylalanine, 0.2~30mg/ml of leucine, color
0.05~30mg/ml of 0.05~20mg/ml of propylhomoserin, 0.05~30mg/ml of threonine and glycine, alanine 0.05~
30mg/ml, 0.05~30mg/ml of serine, 0.05~20mg/ml of 0.05~20mg/ml of aspartic acid, glutamic acid and its amine,
0.05~30mg/ml of proline, 0.05~40mg/ml of arginine, 0.05~30mg/ml of histidine, 0.05~30mg/ of tyrosine
Ml, 0.05~50mg/ml of cystine.
4. cell transport according to claim 2 saves liquid, which is characterized in that it includes following that the cell transport, which saves liquid,
The combination of one or more of vitamin: 0.05~150IU/ml of vitamin A, 0.05~25ug/ml of vitamin B2, dimension
Raw 0.05~500IU/ml of element D, 0.05~30ug/ml of vitamin E, 0.5~300ug/ml of vitamin C.
5. cell transport according to claim 3 saves liquid, which is characterized in that it includes following that the cell transport, which saves liquid,
The combination of one or more of amino acid: methionine 2.5mg/ml, valine 3mg/ml, lysine 5mg/ml, different bright
Propylhomoserin 2mg/ml, phenylalanine-3,4-quinone .5mg/ml, leucine 2mg/ml, tryptophan 2mg/ml, threonine 4mg/ml and glycine
3mg/ml, alanine 3.5mg/ml, serine 1.5mg/ml, aspartic acid 2mg/ml, glutamic acid and its amine 5.5mg/ml, dried meat ammonia
Sour 2.2mg/ml, arginine 4.6mg/ml, histidine 2.5mg/ml, tyrosine 3.5mg/ml, cystine 8.5mg/ml.
6. cell transport according to claim 4 saves liquid, which is characterized in that it includes following that the cell transport, which saves liquid,
The combination of one or more of vitamin: vitamin A 1.25IU/ml, vitamin B2 2.5ug/ml, vitamin D
10IU/ml, vitamin E 7.5ug/ml, vitamin C 25ug/ml.
7. cell transport according to any one of claim 1 to 6 saves liquid, which is characterized in that the cell transport is protected
Liquid storage includes: glucose 0.2mg/ml, sodium chloride 3mg/ml, potassium chloride 1mg/ml, calcium chloride 0.8mg/ml, sodium lactate 8mg/
ml。
8. the application that a kind of transport of cell described in any one of claims 1 to 7 saves liquid characterized by comprising
Cell and cell transport are saved liquid to mix, cell suspension is made；
The cell suspension is moved into container, and keeping the volume ratio of the cell suspension and air is 1:(0.2~30)；
The container is stored at a temperature of 2~15 DEG C.
9. the application that cell transport according to claim 8 saves liquid, which is characterized in that cell is close in the cell suspension
Degree is 1 × 102~1 × 108/ml；
After the cell suspension is moved into the container, keeping the volume ratio of the cell suspension and air is 1:5；
The container is stored at a temperature of 12 DEG C.
10. the application that cell transport according to claim 8 or claim 9 saves liquid, which is characterized in that the method also includes:
When the cell suspension described in clinical application, cell suspension directly is taken out from the container；
It is added in physiological saline, human body is inputted by intravenous injection or other injection systems.
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Cited By (4)
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| CN113100227A (en) * | 2021-03-31 | 2021-07-13 | 北京益华生物科技有限公司 | NK cell transfusion liquid capable of being directly input into human body and preparation method and application thereof |
| CN114600869A (en) * | 2022-03-14 | 2022-06-10 | 北京益华生物科技有限公司 | Cell transportation preservation solution and application thereof |
| CN115315182A (en) * | 2020-03-27 | 2022-11-08 | 上海我武干细胞科技有限公司 | Cell preservation solution and kit thereof |
| CN115885981A (en) * | 2023-03-08 | 2023-04-04 | 北京百奥益康医药科技有限公司 | Preservation solution for maintaining activity of isolated histiocyte and preparation method and application thereof |
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