CN207121593U - A kind of freeze-stored cell kit - Google Patents
A kind of freeze-stored cell kit Download PDFInfo
- Publication number
- CN207121593U CN207121593U CN201720586861.1U CN201720586861U CN207121593U CN 207121593 U CN207121593 U CN 207121593U CN 201720586861 U CN201720586861 U CN 201720586861U CN 207121593 U CN207121593 U CN 207121593U
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- CN
- China
- Prior art keywords
- cell
- kit
- container
- physiological saline
- freeze
- 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.)
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Links
- 238000005138 cryopreservation Methods 0.000 claims abstract description 29
- 239000002504 physiological saline solution Substances 0.000 claims abstract description 24
- 238000002360 preparation method Methods 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 210000004369 blood Anatomy 0.000 claims description 2
- 239000008280 blood Substances 0.000 claims description 2
- 238000000034 method Methods 0.000 abstract description 13
- 230000008569 process Effects 0.000 abstract description 9
- 238000009666 routine test Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 80
- 238000011084 recovery Methods 0.000 description 13
- 206010028980 Neoplasm Diseases 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 238000007710 freezing Methods 0.000 description 6
- 230000008014 freezing Effects 0.000 description 6
- 239000000427 antigen Substances 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 102000036639 antigens Human genes 0.000 description 5
- 238000002659 cell therapy Methods 0.000 description 5
- 238000001514 detection method Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 230000035479 physiological effects, processes and functions Effects 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 239000007788 liquid Substances 0.000 description 3
- 230000004899 motility Effects 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000010257 thawing Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000005484 gravity Effects 0.000 description 2
- 235000015110 jellies Nutrition 0.000 description 2
- 239000008274 jelly Substances 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000009087 cell motility Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 210000004986 primary T-cell Anatomy 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
Abstract
The utility model provides a kind of freeze-stored cell kit, and specifically, described kit includes：Physiological saline container, described physiological saline container are used to deposit physiological saline；Cell cryopreservation container, described cell cryopreservation bag are used to deposit freeze-stored cell preparation；Sterile connector, described disposable sterilized connector include a conduit portion, and are connected to first sting device and the second sting device at conduit portion two.Cryopreservation resuscitation process can be completed by shirtsleeve operation in Routine Test Lab using the kit.
Description
Technical field
The utility model belongs to cell therapy field, specifically, the utility model provide it is a kind of it is disposable can vein drop
The Frozen cells preparation of note, and the Frozen cells preparation is used simplification.
Background technology
When cell therapy is applied, the fresh cell of generally use enters row vein or local infusion, or thin using freezing
Born of the same parents are used after washing.Fresh cell is before Clinical practice, and the term of validity typically only less than 8 hours is unfavorable
In long-range transport, the change of the clinical patient state of an illness can not be adapted to.Therefore, cell, gesture if desired are applied in therapy field
The process for being frozen and being recovered to cell must be related to.
During the cryopreservation resuscitation of laboratory, the operation that recovery washing is carried out to the cell frozen is usually required in cleaning
Carried out in environment (such as in desinfection chamber), and need the biotechnology personnel of specialty to be operated.However, in the process of actual therapeutic
In, the process of cryopreservation resuscitation is carried out in general hospital, can not provide the environment of desinfection chamber, and also lacking has corresponding experiment
The personnel of skill, which has limited promoting the use of for cell therapy.
In summary, this area can carry out cell jelly there is an urgent need to a kind of simple to operate under routine experimentation room environmental
Deposit the method for recovery and corresponding device.
Utility model content
The utility model provides a kind of kit for being applied to carry out cell frozen preparation with disposable frozen stock solution, uses
Described kit, the operation for formula of simply making up a prescription can be used, which to carry out cell recovery and dilution, can both carry out Clinical practice, from
And effectively solves the above mentioned problem in cell therapy field.
First aspect of the present utility model, there is provided a kind of freeze-stored cell kit, the kit include：
Physiological saline container, described physiological saline container are used to deposit physiological saline；
Cell cryopreservation container, described cell cryopreservation bag are used to deposit freeze-stored cell preparation；
Sterile connector, described disposable sterilized connector includes a conduit portion, and is connected to conduit portion
First sting device and the second sting device at two.
In another preference, described sterile connector is disposable sterilized connector.
In another preference, described physiological saline container has one can puncture portion, and the described portion that punctures can be with
Punctured by the first sting device of the sterile connector.
In another preference, described cell cryopreservation container has one can puncture portion, and the described portion that punctures can be with
Punctured by the second sting device of the sterile connector.
In another preference, described physiological saline container is hermetic bag.
In another preference, described cell cryopreservation container is hermetic bag.
In another preference, the first described sting device and the second sting device are selected from the group：Puncture needle, Y types are defeated
Blood device.
In another preference, described sting device also has a sterile sleeve.
In another preference, described kit also includes water bath device.
In another preference, described water bath device is thermostat water bath.
It should be understood that in the scope of the utility model, above-mentioned each technical characteristic of the present utility model and below (such as implementation
Example) in specifically describe each technical characteristic between can be combined with each other, so as to form new or preferable technical scheme.It is limited to
Length, no longer tire out one by one state herein.
Brief description of the drawings
Fig. 1 is the schematic diagram of cell cryopreservation container；
Fig. 2 is disposable sterile connector schematic diagram；
Fig. 3 is physiology saline container schematic diagram.
Embodiment
The present inventor's in-depth study by long-term, devise a kind of easily to operate for cell cryopreservation-recovery
Kit.Described kit can simplify operating procedure so that the conventional cryopreservation resuscitation process energy carried out in an aseptic environment
Enough under routine experimentation room environmental, easily carried out by healthcare givers.Based on above-mentioned discovery, it is new that inventor completes this practicality
Type.
Frozen cells kit
The utility model provides a kind of Frozen cells kit, and described kit includes：
Physiological saline container, described physiological saline container are used to deposit physiological saline；
Cell cryopreservation container, described cell cryopreservation bag are used to deposit freeze-stored cell preparation；
Sterile connector, described disposable sterilized connector includes a conduit portion, and is connected to conduit portion
First sting device and the second sting device at two.
Described physiological saline container and cell cryopreservation container preferably have a portion that can puncture, and the described portion that punctures can be with
Punctured by the first sting device of the sterile connector and the second sting device, so that described physiological saline container and thin
The liquid that born of the same parents are frozen in container interconnects.In preferred embodiment of the present utility model, described physiological saline container and
Cell cryopreservation container is hermetic bag.
In another preference, described physiological saline container and cell cryopreservation volume of a container are equal or approximate, so as to
In to described freeze-stored cell preparation progress 1:1 dilution.
Due to needing transfer liquid, it is therefore preferable that the physiological saline container and cell cryopreservation container are respectively provided with air pressure and put down
Heng Kou.Described air pressure balance mouth can be that envelope has pellicle or the opening of other similar materials, can make physiological saline or jelly
Depositing cell preparation can not flow out, and gas can penetrate.In a kind of preferred embodiment, when container is plastic soft bag, institute
The container stated does not have air pressure balance mouth.
Described sterile connector is preferably disposable sterilized connector, to avoid introducing pollution in operation.
In the preferred embodiments of the present invention, the first described sting device and the second sting device are puncture needle.More preferably implementing
In example, described puncture needle also has a sterile sleeve.
Freeze-stored cell preparation
As used herein, term " freeze-stored cell preparation " and " Frozen cells preparation " are used interchangeably, and are referred both in low temperature
The preparation containing therapeutic activity cell preserved under (at -80 DEG C, or in liquid nitrogen environment) state.
Described freeze-stored cell preparation includes cell and frozen stock solution, and cell can be kept to live under the conditions of cryopreservation
Property.Wherein described cell can be used for the competent cell of cell therapy, and its species has no particular limits, the utility model
Preferred embodiment in, described cell is the cell being selected from the group：T cell, gene after primary T cells, amplification cultivation turn
T cell, stem cell after leading, or its combination.
Cryopreservation method
Using kit of the present utility model, the cell frozen can be recovered under conventional ambient non-sterile, at this
In the preferred embodiment of utility model, described resuscitation process includes step：
(i) described freeze-stored cell preparation is provided；
(ii) described freeze-stored cell preparation is immediately placed in 35-40 DEG C of thermostatic equipment and recovered, recovered
Cell afterwards-frozen stock solution compound；
(iii) cell in the preparation is transferred in injection carrier with syringe or disposable connector.It is excellent
In the mode of operation of choosing, after 75% alcohol clean surface, bag and physiology salt are frozen with the connection of disposable sterile connector
Water bag.
(iv) normal saline bag is raised, makes physiological saline is full to freeze bag under gravity.Normal saline bag is lowerd, is lifted
High cell cryopreservation bag, makes cell suspension flow under the influence of gravity into normal saline bag.
(v) repeat step (iv) twice, extracts connector, makes cell recovery and be diluted in normal saline bag, faced
Bed application.
In another preference, described recovery also includes step：Survival rate test is carried out to cell, if motility rate meets use
It is required that then for Clinical practice.
Described thermostatic equipment has no particular limits, and can be thermostatic equipment (such as water bath with thermostatic control commonly used in laboratory
Pot).
In another preference, described cell thawing time is 30 seconds to 3 minutes.
Compared with prior art, major advantage of the present utility model includes：
It is 1. simple to operate：Only need a thermostat water bath just can complete whole cell recovery without other equipment
Operation, it is not necessary to which professional and technical personnel is operated.Ordinary people is by simple training demonstration with regard to that can carry out cell recovery.
2. cell depletion is low：The cell of defrosting is drawn into physiology salt from freezing by traditional method using syringe in bag
In water bag.This patent can make physiology salt using disposable sterile connector connection normal saline bag and cell cryopreservation bag
Water rinses cell cryopreservation bag repeatedly, so as to reduce the loss in bag.
3. lift the motility rate of recovery cell：During traditional syringe suction of cells, syringe needle is relatively thin sharp to cause cell
Motility rate declines.The speed of operating personnel's suction is grasped the improper Cell viability that also results in and declined.
4. reduce the risk of pollution：Clinical cytology requires sterile more without other pollutions, the mode of traditional syringe suction
The secondary risk that syringe needle exposure can be increased to pollution in atmosphere.This patent uses the disposable sterile connection with double-ended needle
Device, the time of syringe needle exposure is significantly reduced, so as to reduce the risk of pollution.
With reference to specific embodiment, the utility model is expanded on further.It should be understood that these embodiments are merely to illustrate this
Utility model rather than limitation the scope of the utility model.The experimental method of unreceipted actual conditions in the following example, lead to
Often according to normal condition, or according to the condition proposed by manufacturer.Unless otherwise indicated, otherwise percentage and number be by weight
Calculate.
Embodiment 1
Frozen cells preparation freezes container to freeze bag.Cell type is CAR-T cells.Freeze specification 2 × 107/
10ml.The component of Frozen cells preparation is as shown in the table.After described CAR-T cells and 100ml are frozen into 3 months, at 37 DEG C
Thawed 2 minutes in water-bath, the cell of defrosting is diluted to by 100ml physiological saline using kit such as described in the utility model
In.The cell of dilution is taken to carry out survival rate test, tumour antigen response detection and ability of cell proliferation detection.
Cell viability changes：The cell suspension of dilution carries out survival rate test with the method for Trypan Blue, as a result as in Fig. 1
It is shown.As a result show, cell maintains higher survival rate after recovery.
Freeze the multiplication capacity of rear cell：The cell of cryopreservation resuscitation is placed in complete medium and continues to cultivate, daily note
Record the density of cell.Every 2-3 days supplementing culture mediums, growth curve (as shown in Figure 2) is drawn.It can be seen that freezing
After resuscitation process, cell still has preferable multiplication capacity.
Freeze the tumour antigen responsibility of front and rear CAR-T cells：The cell of cryopreservation resuscitation and swelling for carrying tumour antigen
Oncocyte 1:1 co-cultures the IFNr amounts for overnight, collecting culture supernatant detection CAR-T cell releases；Collect cell detection CD3 sun
Property cell expression CD137 ratio assess tumour antigen responsibility.Data before freezing and continue culture 7 days after freezing
As a comparison, comparison diagram is as shown in Figure 3 for data.As a result show, experienced freeze-resuscitation process after, the tumour of cell resists
Former responsibility is not decreased obviously.
Conclusion：The Frozen cells preparation motility rate for freezing recovery in 3 months reaches more than 90%.The frost CAR-T cells of recovery
Preparation still has competence for added value.The frost CAR-T cell preparations of recovery, the reversible decline of tumour antigen responsibility occurs,
It is restored after continuing culture.Illustrate, by saving the original efficiency of fresh cells after the cryopreservation resuscitation process, again to face
Bed application.
All it is incorporated as referring in this application in all documents that the utility model refers to, just as each document quilt
It is individually recited as with reference to such.In addition, it is to be understood that after above-mentioned instruction content of the present utility model has been read, this area skill
Art personnel can make various changes or modifications to the utility model, and these equivalent form of values equally fall within the application appended claims
Book limited range.
Claims (9)
- A kind of 1. freeze-stored cell kit, it is characterised in that including：Physiological saline container, described physiological saline container are used to deposit physiological saline；Cell cryopreservation container, described cell cryopreservation bag are used to deposit freeze-stored cell preparation；Sterile connector, described sterile connector includes a conduit portion, and is connected to the first of conduit portion two Sting device and the second sting device.
- 2. kit as claimed in claim 1, it is characterised in that described physiological saline container has one can puncture portion, and The described portion that punctures can be by the first sting device puncture of the sterile connector.
- 3. kit as claimed in claim 1, it is characterised in that described cell cryopreservation container has one can puncture portion, and The described portion that punctures can be by the second sting device puncture of the sterile connector.
- 4. kit as claimed in claim 1, it is characterised in that described physiological saline container is hermetic bag.
- 5. kit as claimed in claim 1, it is characterised in that described cell cryopreservation container is hermetic bag.
- 6. kit as claimed in claim 1, it is characterised in that described the first sting device and the second sting device is selected from The following group：Puncture needle, Y-shaped blood transfusion apparatus.
- 7. kit as claimed in claim 6, it is characterised in that described sting device also has a sterile sleeve.
- 8. kit as claimed in claim 1, it is characterised in that described kit also includes water bath device.
- 9. kit as claimed in claim 8, it is characterised in that described water bath device is thermostat water bath.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720586861.1U CN207121593U (en) | 2017-05-24 | 2017-05-24 | A kind of freeze-stored cell kit |
| PCT/CN2018/088245 WO2018214942A1 (en) | 2017-05-24 | 2018-05-24 | Cell cryopreservation kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720586861.1U CN207121593U (en) | 2017-05-24 | 2017-05-24 | A kind of freeze-stored cell kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN207121593U true CN207121593U (en) | 2018-03-20 |
Family
ID=61613909
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201720586861.1U Active CN207121593U (en) | 2017-05-24 | 2017-05-24 | A kind of freeze-stored cell kit |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN207121593U (en) |
| WO (1) | WO2018214942A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018214942A1 (en) * | 2017-05-24 | 2018-11-29 | 西比曼生物科技（上海）有限公司 | Cell cryopreservation kit |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN202170342U (en) * | 2011-08-02 | 2012-03-21 | 江苏省北科生物科技有限公司 | Cord blood separation bigeminal bag |
| CN102586104B (en) * | 2011-12-23 | 2013-09-25 | 北京瑞建天行生物技术有限公司 | Multifunctional container for freezing cells and resurrecting frozen cells |
| CN205473713U (en) * | 2016-02-26 | 2016-08-17 | 博雅干细胞科技有限公司 | Be fit for dry cell cryopreserving bag that cell feedback was used |
| CN207121593U (en) * | 2017-05-24 | 2018-03-20 | 西比曼生物科技（上海）有限公司 | A kind of freeze-stored cell kit |
-
2017
- 2017-05-24 CN CN201720586861.1U patent/CN207121593U/en active Active
-
2018
- 2018-05-24 WO PCT/CN2018/088245 patent/WO2018214942A1/en active Application Filing
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018214942A1 (en) * | 2017-05-24 | 2018-11-29 | 西比曼生物科技（上海）有限公司 | Cell cryopreservation kit |
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| Publication number | Publication date |
|---|---|
| WO2018214942A1 (en) | 2018-11-29 |
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|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20180820 Address after: Hongkong Jinzhong Chinese Cotton Tree Drive Fairmont House No. 8 Building 402 room 4 Patentee after: CELLULAR BIOMEDICINE GROUP HK Ltd. Address before: No. 5, building No. 1, No. 333, Guiping Road, Xuhui District, Shanghai Co-patentee before: CELLULAR BIOMEDICINE GROUP (WUXI) Ltd. Patentee before: CELLULAR BIOMEDICINE GROUP (SHANGHAI) Ltd. |
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| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20221025 Address after: Floor 6, Building 1, No. 333 Guiping Road, Xuhui District, Shanghai Patentee after: CELLULAR BIOMEDICINE GROUP (SHANGHAI) Ltd. Address before: Hongkong Jinzhong Chinese Cotton Tree Drive Fairmont House No. 8 Building 402 room 4 Patentee before: CELLULAR BIOMEDICINE GROUP HK Ltd. |
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| TR01 | Transfer of patent right |
Effective date of registration: 20221223 Address after: 201210 Building 3, No. 85 Faraday Road, Pudong New Area (Shanghai) Pilot Free Trade Zone, Shanghai Patentee after: SHANGHAI CELLULAR BIOPHARMACEUTICAL Group Ltd. Address before: 200233 6th floor, building 1, 333 Guiping Road, Xuhui District, Shanghai Patentee before: CELLULAR BIOMEDICINE GROUP (SHANGHAI) Ltd. |