CN106801067A - A kind of preparation method of Chimeric antigen receptor T cell - Google Patents
A kind of preparation method of Chimeric antigen receptor T cell Download PDFInfo
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- CN106801067A CN106801067A CN201611220109.1A CN201611220109A CN106801067A CN 106801067 A CN106801067 A CN 106801067A CN 201611220109 A CN201611220109 A CN 201611220109A CN 106801067 A CN106801067 A CN 106801067A
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Abstract
The invention discloses a kind of preparation method of Chimeric antigen receptor T cell, comprise the following steps：S1, centrifuge tube is transferred to by blood, with normal saline dilution, is well mixed；S2, the cardiografin of 3mL is added into Ficoll solution, obtain mixed liquor, mixed liquor is put into centrifuge tube, and centrifuge tube is placed on operating desk vertically, the blood that will have been diluted is put into 1/2nd, and be slowly stirred, after static 1 2min, remaining 1/2nd blood is added, be slowly stirred until well mixed；S3, will add the centrifuge tube of good blood, cardiografin and Ficoll solution to be placed on centrifuge in S2,25 45min are centrifuged at a temperature of 20 30 DEG C；S4, operating desk is moved to by the centrifuge tube after being centrifuged in S3, is taken the tunica albuginea layer i.e. buffy coat of centre and is transferred on culture plate.Chimeric antigen receptor T cell prepared by the present invention has certain functional activity, can provide technique direction for oncotherapy.
Description
Technical field
The present invention relates to gene engineering technology field, more particularly to a kind of preparation method of Chimeric antigen receptor T cell.
Background technology
Cancer is always to perplex the huge difficult problem of the mankind.Newly hair tumor cases are about 3,120,000 every year for China, average daily
8550 people mean it is per minute just there are 6 people to be just diagnosed as malignant tumour, these data all show that cancer is still what is posed a health risk
One big killer.For many years, the method for the treatment of of cancer is mainly operation, chemotherapy and radiation, and the clinical treatment to tumor patient is effective
Rate is still very low, and according to statistics, the effective percentage of same antineoplastic is only 25%.For same tumour, with same medicine,
The treatment method of standard dose, the curative effect risen with different patients and toxic and side effect but have very big difference.Cause this
Result main reason is that, tumour is a kind of multi-pathogenesis, heterogeneous disease.Over the past decade, individual patients are depended on to give birth to
The tumour personalized treatment of thing mark is gradually replacing according to the Therapeutic mode of Conventional wisdom, and shows superior curative effect
With wide prospect, such as Imatinib and Herceptin.
Immunotherapy is, after a kind of novel therapies occurred after operation, chemotherapy, radiotherapy and targeted therapies, to be referred to as treating cancer
" the fifth-largest therapy " of disease.It is to resist cancer using the strength of patient's self immune system；Immunotherapy for cancer is mainly wrapped
Include adoptive cell therapy, immunomodulator, tumor vaccine and immune binding site blocking treatment etc..Although relevant immunization therapy
Research just carried out early in before 30 years, nearest 2 years《Cell》、《Science》、《It is natural》Multinomial being somebody's turn to do is published etc. international top periodical
The breakthrough achievement in research in field, pharmacy giant also constantly releases the heavy pound therapy of this type.In December, 2014 No. 6 to No. 9 is old
In United States blood association annual meeting (ASH) that Kingsoft is held, immunotherapy for cancer is to have earned enough eyeball, and CAR-T therapies are even more
Receive much concern.
The concept of CART is that Israel doctor ZeligEshhar proposes the eighties in last century, 1989 develop first it is embedding
Close antigen receptor T cell.CART therapies are Chimeric antigen receptor T cell therapy, have bypassed PD therapies and have started adoptive immunity response
This difficulty, has started the medical history new page of great-leap-forward development.CART is directed to that less reliable adoptive immunity, introduces complete
New concept and method, successfully induces the adoptive immunity response of anticancer.CART first solves the problem of antigen.Since
Immune system can not find antigen under natural situation, and that just artificially specifies a kind of antigen.Investigate acute lymphoblastic B cell leukemia
CART therapies.There is a differentiation antigen CD19 on lymphocytic B cells surface, and either normal B cells or canceration B cell have table
Reach.CD19 is selected to may insure no fish that has escape the net for target antigen.Next, how to ensure that T cell can find CD19 target antigensThis
It is also the core technology of CART.Natural T cell certainly will not be by CD19 as target antigen.CD19 is expressed in normal B cells,
T cell is tolerated to CD19.One antigen receptor is fitted to T cell surface by CART.This antigen receptor essence is a kind of antibody, right
CD19 has compatibility very high.Both encounter and will combine, and do not abandon.Then, coupled with antibody protein and positioned, T cell is just
Cancer cell is pegged.So there is a metaphor, this antigen receptor gives T cell navigation just as GPS, and guarantee is provided to find target.
Antibody how is inserted into T cellThis is realized by Gene transfer techniques.Here transfect be not antibody in itself
But the gene of encoding antibody, that is to say, that transfection is DNA.Then the DNA meeting encoding antibodies of transfection are expressed in T cell table
Face.Existing Chimeric antigen receptor T cell makes nonstandard, relatively complicated, without procedure operation, be this we have proposed
A kind of preparation method of Chimeric antigen receptor T cell.
The content of the invention
Based on the technical problem that background technology is present, the present invention proposes a kind of preparation side of Chimeric antigen receptor T cell
Method.
A kind of preparation method of Chimeric antigen receptor T cell proposed by the present invention, comprises the following steps：
S1, centrifuge tube is transferred to by blood, with normal saline dilution, is well mixed；
S2, Ficoll solution is added by the cardiografin of 3mL, obtains mixed liquor, and mixed liquor is put into centrifuge tube, and will
Centrifuge tube is placed on operating desk vertically, and the blood that will have been diluted is put into 1/2nd, and is slowly stirred, after static 1-2min,
Remaining 1/2nd blood is added, is slowly stirred until well mixed；
S3, will add the centrifuge tube of good blood, cardiografin and Ficoll solution to be placed on centrifuge, at 20-30 DEG C in S2
At a temperature of 25-45min is centrifuged；
S4, operating desk is moved to by the centrifuge tube after being centrifuged in S3, is taken the tunica albuginea layer i.e. buffy coat of centre and is transferred to
On culture plate；
S5, culture plate is placed in incubator and cultivates 24-48h；
S6, adds virus on the culture plate after being cultivated in S5, and is put into incubator to continue to cultivate and continues 5-10 days；
S7, the cell on detection culture plate, and determine whether T cell is infected, you can complete Chimeric antigen receptor T cell
Preparation.
Preferably, the physiological saline and the volume ratio of blood are 1：1-2.
Preferably, the physiological saline and the volume ratio of blood are 1：2.
Preferably, the mass-volume concentration of the cardiografin is 340g/L.
Preferably, in the S3, centrifugation is placed on by the centrifuge tube of good blood, cardiografin and Ficoll solution is added in S2
Machine, 35min is centrifuged at a temperature of 25 DEG C.
Preferably, in the S4, operating desk is ultra-clean operating desk.
Preferably, in the S5, the temperature of incubator is 36-38 DEG C, and the carbon dioxide in incubator is 4-10%.
Preferably, in the S5, the temperature of incubator is 37 DEG C, and the carbon dioxide in incubator is 7%.
In the present invention, blood is first transferred to centrifuge tube, with normal saline dilution, be well mixed, cardiografin is added
Ficoll solution, obtains mixed liquor, and mixed liquor is put into centrifuge tube, and the blood that will have been diluted is put into 1/2nd, and delays
Slow stirring, after static 1-2min, adds remaining 1/2nd blood, is slowly stirred until well mixed, will plus good blood,
Cardiografin is placed on centrifuge with the centrifuge tube of Ficoll solution, and 25-45min is centrifuged at a temperature of 20-30 DEG C, will be centrifuged
Centrifuge tube afterwards moves to operating desk, takes the tunica albuginea layer i.e. buffy coat of centre and is transferred on culture plate, and culture plate is placed
24-48h is cultivated in incubator, virus will be added on the culture plate after culture, and be put into the lasting 5- of continuation culture in incubator
10 days, the cell on detection culture plate, and determine whether T cell is infected, you can complete the system of Chimeric antigen receptor T cell
Standby, Chimeric antigen receptor T cell prepared by the present invention has certain functional activity, can provide technical side for oncotherapy
To.
Specific embodiment
The present invention is made with reference to specific embodiment further explain.
Embodiment one
A kind of preparation method of Chimeric antigen receptor T cell proposed by the present invention, comprises the following steps：
S1, centrifuge tube is transferred to by blood, with normal saline dilution, is well mixed；
S2, Ficoll solution is added by the cardiografin of 3mL, obtains mixed liquor, and mixed liquor is put into centrifuge tube, and will
Centrifuge tube is placed on operating desk vertically, and the blood that will have been diluted is put into 1/2nd, and is slowly stirred, after static 1min, then
Remaining 1/2nd blood is added, is slowly stirred until well mixed；
S3, will add the centrifuge tube of good blood, cardiografin and Ficoll solution to be placed on centrifuge, in 20 DEG C of temperature in S2
The lower centrifugation 25min of degree；
S4, operating desk is moved to by the centrifuge tube after being centrifuged in S3, is taken the tunica albuginea layer i.e. buffy coat of centre and is transferred to
On culture plate；
S5, culture plate is placed in incubator and cultivates 24h；
S6, adds virus on the culture plate after being cultivated in S5, and is put into incubator and continues culture and continue 5 days；
S7, the cell on detection culture plate, and determine whether T cell is infected, you can complete Chimeric antigen receptor T cell
Preparation.
Embodiment two
A kind of preparation method of Chimeric antigen receptor T cell proposed by the present invention, comprises the following steps：
S1, centrifuge tube is transferred to by blood, with normal saline dilution, is well mixed；
S2, Ficoll solution is added by the cardiografin of 3mL, obtains mixed liquor, and mixed liquor is put into centrifuge tube, and will
Centrifuge tube is placed on operating desk vertically, and the blood that will have been diluted is put into 1/2nd, and is slowly stirred, and static 1.After 2min,
Remaining 1/2nd blood is added, is slowly stirred until well mixed；
S3, will add the centrifuge tube of good blood, cardiografin and Ficoll solution to be placed on centrifuge, in 22 DEG C of temperature in S2
The lower centrifugation 30min of degree；
S4, operating desk is moved to by the centrifuge tube after being centrifuged in S3, is taken the tunica albuginea layer i.e. buffy coat of centre and is transferred to
On culture plate；
S5, culture plate is placed in incubator and cultivates 30h；
S6, adds virus on the culture plate after being cultivated in S5, and is put into incubator and continues culture and continue 6 days；
S7, the cell on detection culture plate, and determine whether T cell is infected, you can complete Chimeric antigen receptor T cell
Preparation.
Embodiment three
A kind of preparation method of Chimeric antigen receptor T cell proposed by the present invention, comprises the following steps：
S1, centrifuge tube is transferred to by blood, with normal saline dilution, is well mixed；
S2, Ficoll solution is added by the cardiografin of 3mL, obtains mixed liquor, and mixed liquor is put into centrifuge tube, and will
Centrifuge tube is placed on operating desk vertically, and the blood that will have been diluted is put into 1/2nd, and is slowly stirred, after static 1.5min,
Remaining 1/2nd blood is added, is slowly stirred until well mixed；
S3, will add the centrifuge tube of good blood, cardiografin and Ficoll solution to be placed on centrifuge, in 25 DEG C of temperature in S2
The lower centrifugation 35min of degree；
S4, operating desk is moved to by the centrifuge tube after being centrifuged in S3, is taken the tunica albuginea layer i.e. buffy coat of centre and is transferred to
On culture plate；
S5, culture plate is placed in incubator and cultivates 36h；
S6, adds virus on the culture plate after being cultivated in S5, and is put into incubator and continues culture and continue 7 days；
S7, the cell on detection culture plate, and determine whether T cell is infected, you can complete Chimeric antigen receptor T cell
Preparation.
Example IV
A kind of preparation method of Chimeric antigen receptor T cell proposed by the present invention, comprises the following steps：
S1, centrifuge tube is transferred to by blood, with normal saline dilution, is well mixed；
S2, Ficoll solution is added by the cardiografin of 3mL, obtains mixed liquor, and mixed liquor is put into centrifuge tube, and will
Centrifuge tube is placed on operating desk vertically, and the blood that will have been diluted is put into 1/2nd, and is slowly stirred, after static 1.8min,
Remaining 1/2nd blood is added, is slowly stirred until well mixed；
S3, will add the centrifuge tube of good blood, cardiografin and Ficoll solution to be placed on centrifuge, in 28 DEG C of temperature in S2
The lower centrifugation 40min of degree；
S4, operating desk is moved to by the centrifuge tube after being centrifuged in S3, is taken the tunica albuginea layer i.e. buffy coat of centre and is transferred to
On culture plate；
S5, culture plate is placed in incubator and cultivates 42h；
S6, adds virus on the culture plate after being cultivated in S5, and is put into incubator and continues culture and continue 8 days；
S7, the cell on detection culture plate, and determine whether T cell is infected, you can complete Chimeric antigen receptor T cell
Preparation.
Embodiment five
A kind of preparation method of Chimeric antigen receptor T cell proposed by the present invention, comprises the following steps：
S1, centrifuge tube is transferred to by blood, with normal saline dilution, is well mixed；
S2, Ficoll solution is added by the cardiografin of 3mL, obtains mixed liquor, and mixed liquor is put into centrifuge tube, and will
Centrifuge tube is placed on operating desk vertically, and the blood that will have been diluted is put into 1/2nd, and is slowly stirred, after static 2min, then
Remaining 1/2nd blood is added, is slowly stirred until well mixed；
S3, will add the centrifuge tube of good blood, cardiografin and Ficoll solution to be placed on centrifuge, in 30 DEG C of temperature in S2
The lower centrifugation 45min of degree；
S4, operating desk is moved to by the centrifuge tube after being centrifuged in S3, is taken the tunica albuginea layer i.e. buffy coat of centre and is transferred to
On culture plate；
S5, culture plate is placed in incubator and cultivates 48h；
S6, adds virus on the culture plate after being cultivated in S5, and is put into incubator and continues culture and continue 10 days；
S7, the cell on detection culture plate, and determine whether T cell is infected, you can complete Chimeric antigen receptor T cell
Preparation.
Chimeric antigen receptor T cell prepared by the present invention has certain functional activity, can provide skill for oncotherapy
Art direction.
The above, the only present invention preferably specific embodiment, but protection scope of the present invention is not limited thereto,
Any one skilled in the art the invention discloses technical scope in, technology according to the present invention scheme and its
Inventive concept is subject to equivalent or change, should all be included within the scope of the present invention.
Claims (8)
1. a kind of preparation method of Chimeric antigen receptor T cell, it is characterised in that comprise the following steps：
S1, centrifuge tube is transferred to by blood, with normal saline dilution, is well mixed；
S2, Ficoll solution is added by the cardiografin of 3mL, obtains mixed liquor, and mixed liquor is put into centrifuge tube, and will centrifugation
Pipe is placed on operating desk vertically, and the blood that will have been diluted is put into 1/2nd, and is slowly stirred, and after static 1-2min, then is added
Enter remaining 1/2nd blood, be slowly stirred until well mixed；
S3, will add the centrifuge tube of good blood, cardiografin and Ficoll solution to be placed on centrifuge, in 20-30 DEG C of temperature in S2
The lower centrifugation 25-45min of degree；
S4, operating desk is moved to by the centrifuge tube after being centrifuged in S3, is taken the tunica albuginea layer i.e. buffy coat of centre and is transferred to culture
On plate；
S5, culture plate is placed in incubator and cultivates 24-48h；
S6, adds virus on the culture plate after being cultivated in S5, and is put into incubator to continue to cultivate and continues 5-10 days；
S7, the cell on detection culture plate, and determine whether T cell is infected, you can complete the system of Chimeric antigen receptor T cell
It is standby.
2. a kind of preparation method of Chimeric antigen receptor T cell according to claim 1, it is characterised in that the physiology
Salt solution is 1 with the volume ratio of blood：1-2.
3. a kind of preparation method of Chimeric antigen receptor T cell according to claim 2, it is characterised in that the physiology
Salt solution is 1 with the volume ratio of blood：2.
4. a kind of preparation method of Chimeric antigen receptor T cell according to claim 1, it is characterised in that the general shadow
The mass-volume concentration of Portugal's amine is 340g/L.
5. the preparation method of a kind of Chimeric antigen receptor T cell according to claim 1, it is characterised in that in the S3,
The centrifuge tube of good blood, cardiografin and Ficoll solution will be added to be placed on centrifuge in S2, be centrifuged at a temperature of 25 DEG C
35min。
6. the preparation method of a kind of Chimeric antigen receptor T cell according to claim 1, it is characterised in that in the S4,
Operating desk is ultra-clean operating desk.
7. the preparation method of a kind of Chimeric antigen receptor T cell according to claim 1, it is characterised in that in the S5,
The temperature of incubator is 36-38 DEG C, and the carbon dioxide in incubator is 4-10%.
8. the preparation method of a kind of Chimeric antigen receptor T cell according to claim 7, it is characterised in that in the S5,
The temperature of incubator is 37 DEG C, and the carbon dioxide in incubator is 7%.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102876631A (en) * | 2012-10-09 | 2013-01-16 | 博雅干细胞科技有限公司 | Method for separating immune cells from blood and application of method to disease treatment |
| CN105349489A (en) * | 2015-12-07 | 2016-02-24 | 广州赛莱拉干细胞科技股份有限公司 | Culture method of CIK cell |
| CN105640991A (en) * | 2016-01-06 | 2016-06-08 | 奥思达干细胞有限公司 | CAR-T cell preparation for treating prostatic cancer and preparation method thereof |
| CN105861531A (en) * | 2016-04-21 | 2016-08-17 | 汪治宇 | Chimeric antigen receptor T cell and preparation method thereof |
-
2016
- 2016-12-26 CN CN201611220109.1A patent/CN106801067A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102876631A (en) * | 2012-10-09 | 2013-01-16 | 博雅干细胞科技有限公司 | Method for separating immune cells from blood and application of method to disease treatment |
| CN105349489A (en) * | 2015-12-07 | 2016-02-24 | 广州赛莱拉干细胞科技股份有限公司 | Culture method of CIK cell |
| CN105640991A (en) * | 2016-01-06 | 2016-06-08 | 奥思达干细胞有限公司 | CAR-T cell preparation for treating prostatic cancer and preparation method thereof |
| CN105861531A (en) * | 2016-04-21 | 2016-08-17 | 汪治宇 | Chimeric antigen receptor T cell and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 金伯泉: "《细胞和分子免疫学实验技术》", 31 December 2002 * |
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