CN106754700A - A kind of preparation method of antitumor adoptive immunity competent cell CIK - Google Patents
A kind of preparation method of antitumor adoptive immunity competent cell CIK Download PDFInfo
- Publication number
- CN106754700A CN106754700A CN201611204772.2A CN201611204772A CN106754700A CN 106754700 A CN106754700 A CN 106754700A CN 201611204772 A CN201611204772 A CN 201611204772A CN 106754700 A CN106754700 A CN 106754700A
- Authority
- CN
- China
- Prior art keywords
- cell
- cik
- preparation
- final concentration
- pbmc
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0646—Natural killers cells [NK], NKT cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2302—Interleukin-2 (IL-2)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2306—Interleukin-6 (IL-6)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2321—Interleukin-21 (IL-21)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2327—Interleukin-27 (IL-27)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/24—Interferons [IFN]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/998—Proteins not provided for elsewhere
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of preparation method of antitumor adoptive immunity competent cell CIK, it is low to overcome the ability of cell proliferation that CIK cell prepared by prior art is present, the problems such as effector cell's group's limited amount and cell culture period long of acquisition.CIK cell prepared by the present invention processes CIK progenitor cells by the use in conjunction of L-6 and IL-21, reduce Treg cell quantities, the relative quantity and tumor-killing ability that improve effector cell, then replace CD3mAb beneficial to TG, targetedly improve amplification, differentiation and the maturation of CD3+CD56+ cell masses.And the TG of clinical practice rank is current CIK preparation systems in the optimization of safety and efficacy and perfect.The last present invention is more suitable for the application of clinical treatment by IL-27 and IL-2 Combined culture CIK cells, the shortening for further increasing the quantity and efficiency of final prepared CIK cell, the adverse reactions of patients that reduction is caused by cell factor, and CIK cell cultivation cycle.
Description
Technical field
The invention belongs to biological technical field, and in particular to a kind of high proliferation power, the antitumor of cytotoxic activity high is adopted
The preparation method of immunocompetent cell CIK.
Background technology
Recently as the progress of basic medical research, people deepen continuously to tumour understanding, and oncotherapy achieves one
Fixed achievement.Traditional operative treatment and radiotherapy, chemotherapy are the three big conventional methods for treating tumour, and biological therapy is then with existing
Based on for front line sciences such as molecular biology, immunology and cell biologies, by immune, neuroendocrine, gene expression,
The links such as angiogenesis adjust the biologically of body itself, directly or indirectly suppress tumour or mitigate treatment-related bad
Reaction.Immunocyte treatment of adopting belongs to the category of tumor biotherapy, mainly by being transfused immunocompetent cell, enhancing tumour
Patient immune function reaches antitumous effect.Cytokine induced kill cell (cytokine induced killer
Cells, CIK cell) it is one of main cell currently used for tumour adoptive immunity infusion of therapeutic.CIK cell be one group simultaneously
Expression CD3+CD56+The T cells group of two kinds of membrane protein molecules, be also called NK (natural killer cell,
NK) sample T lymphocytes, with the powerful anti-tumor activity of T lymphocytes, and NK cells non-principal histocompatibility complex
(major histocompatibility complex, MHC) is restricted to kill knurl advantage.The cell factor of various combination can induce
Go out various CIK cells, current CD3 monoclonal antibodies (CD3 monoclonal antibody, CD3 mAb) and IFN-γ
(Interferon-γ, IFN-γ), frequently as the necessary factor of inducing component, other conventional factors have：IL-2, phytohemagglutin phytolectin
(Phytohemagglutinin, PHA), IL-7, IL-12 etc..Induced by the factor and cellar culture, the CD3 in peripheral blood+
CD56+Cell can be expanded, and be fed back to rear dissolvable kinds of tumor cells in tumor patient body.And CIK has siberian crabapple
The adjustment effect of system, can also improve the quality of life of patient while tumor focus are removed, elongated strap knurl life span, therefore
CIK cell is in antitumor adoptive cellular immunotherapy using relatively broad.
The quantity and function of immunocyte are the keys of clinical adoptive immunity cell therapy, while substantial amounts of clinical evidence
The rise for showing CD4+CD25+ regulatory T cells (regulatory T cells, Treg) quantity and function is to cause immune suppression
System and the key factor of antineoplastic immune failure.Therefore, the plan of Treg cells is targetted in tumour adoptive immunity cell therapy
Slightly such as artificial removal Treg cells, suppression mononuclearcell turn into following pole to Treg cell transformations and suppression Treg functions
Promising therapeutic strategy.CIK cell multiplication capacity obtained by CIK cell preparation method traditional at present is relatively low, acquisition
Effector cell's group's limited amount, and cell culture period is long, therapeutic effect is not satisfactory, is also unfavorable for that carrying out clinic on a large scale pushes away
Extensively.
The content of the invention
The invention provides a kind of high proliferation power, the antitumor adoptive immunity competent cell of cytotoxic activity (efficient) high
The preparation method of CIK, the ability of cell proliferation for overcoming CIK cell presence prepared by prior art is low, the effector cell of acquisition
The problems such as group's limited amount and cell culture period long.
The present invention is adopted the following technical scheme that：
A kind of preparation method of antitumor adoptive immunity competent cell CIK, it is characterised in that comprise the following steps：
1) collection and preparation of PMNC (PBMC)：
1.1) by venipuncture, liquaemin (40,000 U/L) anti-freezing collection peripheral blood 50-100ml；In in superclean bench
By blood sample and physiological saline with 1:1 volume ratio mixed diluting, human peripheral lymphocyte separating liquid (FICOLL configurations) point
From PBMC, lymphocyte separation medium is with dilution blood sample with 1:2 volume ratios add centrifuge tube, at room temperature, 2000-2500rpm centrifugations
20‐30min；
1.2) collected after centrifugation tunica albuginea layer, i.e. PBMC parts；Volume is to 45-50ml and mixes to add physiological saline adjustment,
PBMC is obtained after 1500-1800rpm is centrifuged 5-10min, above step repeated washing cell 2-3 times；
2) preparation of CIK cell：
2.1) adjustment cell density is 1 × 106/ml‐2×106/ ml, is inoculated in containing final concentration of 75-150ng/ml IL-6
And in the CIK basal mediums of 75-150ng/ml IL-21,37 DEG C, be routinely incubated in 5%CO2 incubators；
2.2) after 18-24h, cell is moved into the CIK basal mediums containing final concentration of 850-1200U/ml IFN-γs
Middle cellar culture；
2.3) after 18-24h, to adding final concentration of 350-650ng/ml people's thymus gland globulin 500- in cultivating system
650U/ml IL-2 and 5.5-13.5ng/ml IL-27；
2.4) hereafter final concentration 500-650U/ml IL-2 and 5.5-13.5ng/ml IL- are contained every 3-4d supplement equivalent
In 27 CIK basal mediums, 9-11d is persistently cultivated.Can obtain high proliferation power, the antitumor adoptive immunity of cytotoxic activity high
Competent cell CIK.
Further, the CIK cell basal medium autologous plasmas of 5-13ml/L containing final concentration in the step (2.1)
Serum free medium, specially modified form RPMI-1640 culture mediums.
Step (2) of the present invention is to prepare high proliferation power, the committed step of cytotoxic activity CIK cell high.
Treg cells suppress and antineoplastic immune failure to immunity of organism during the step 2.1 predominantly solves immunization therapy
Technical problem.IL-6 and IL-21 are used in combination PBMC is pre-processed, the two beneficial benefits of aspect can be reached：First,
Can suppress in CIK preparation process, differentiation from mononuclearcell to Treg cells, Treg cells contain in reducing the cultivating system
Amount, second, the ratio of CIK cell quantity and effector cell CD3+CD56+ can be increased, strengthens its killing ability to tumour.IL‐
6 and IL-21 is the important immune-regulating factor of body.IL-6 is main to be produced by fibroblast, endothelial cell and lymphocyte
It is raw, the generation of Treg cells can be suppressed, immunosuppressive action of the Treg cells to body is released, while it is thin to may additionally facilitate CIK
The killing activity of the propagation of born of the same parents and enhancing to tumour cell.The main CD4+T cells by activating of IL-21 are produced, and be may interfere with
The inductive formation process of Treg cells, reduces the generation of Treg cells and then releases its rejection ability to immunocyte and to machine
The inhibitory action of body immune state, strengthens the antineoplastic immune of human body.Therefore the present invention is by IL-21 and IL-6 use in conjunction, one
Aspect can further improve CIK effector cell's anti-tumor immunotherapy effect by reducing Treg cell proportions, on the other hand also
The proliferative ability of CIK cell cultivating system cell and its ability of killing tumor cell can be significantly increased.
The step 2.2 user thymus gland globulin TG replaces current conventional use of monoclonal antibody CD3mAb.It is described
TG is mainly used in preventing and (or) treating kidney transplantation exclusion reaction in clinic, is a kind of have to kinds of surface antigen in immune system
There is specific immunosupress class medicine.This laboratory previous experiments find, TG compared with CD3mAb, raising that can be more special
The amplification of CD3+CD56+ cell masses, differentiation and ripe, the increase of effector cell's ratio can further lift anti-tumor activity.
And it is used for the medicine of clinical treatment as a kind of approved, TG securities are higher, therefore can be used as raising CIK quantity and cell
The prioritization scheme of cytotoxic activity.
The step 2.3 reduces the dosage of IL-2 by Low dose IL -27 and the use in conjunction of IL-2, promotes CIK
Cell maturation, shortens CIK cell cultivation cycle.To reach three beneficial outcomes：1) toxic and side effects of the IL-2 to patient is reduced；2)
Reduce because of the increase of heavy dose of Treg cell masses brought using IL-2；3) shorten the manufacturing cycle of CIK cell, promote antitumor
The clinical large-scale promotion of adoptive immunity competent cell CIK.
It is cell factor of the indispensable promotion lymphopoiesis with survival in cultivating system that IL-2 is, but is had
Research shows that IL-2 can equally effectively facilitate the generation of inductivity Treg cells.Heavy dose IL-2 in the preparation process of CIK cell
Continuous action can cause the Secondary cases of Treg cells to produce so that suppressing CIK cell to a certain extent kills tumor activity.IL‐
27 monocytes for being mainly derived from activation and BMDC, can promote CIK cell to breed and killing ability.Therefore, we
By IL-27 and IL-2 Combined cultures, the beneficial outcomes of following three aspect can be mainly brought：Reduce heavy dose and use cell factor
The cellulotoxic side effect and sufferer adverse reaction rate for bringing；The CIK cell of sufficient amount and function can be obtained；Shorten CIK
Cell culture period, promotes CIK cell ripe, beneficial to later phase clinical large-scale promotion application.
In sum, the CIK cell that prepared by the present invention processes CIK progenitor cells by the use in conjunction of L-6 and IL-21,
Treg cell quantities are reduced, the relative quantity and tumor-killing ability that improve effector cell then replaces CD3 beneficial to TG
MAb, targetedly improves amplification, differentiation and the maturation of CD3+CD56+ cell masses.And the TG of clinical practice rank is current
CIK preparation systems are in the optimization of safety and efficacy and perfect.The last present invention is by IL-27 and IL-2 Combined cultures CIK
Cell, further increases the quantity and efficiency of final prepared CIK cell, and reduction is bad anti-by the patient that cell factor causes
Should, and the shortening of CIK cell cultivation cycle is more suitable for the application of clinical treatment.
Technical solution of the present invention is described further below in conjunction with example.
Brief description of the drawings
Fig. 1 is that the CIK cell for preparing of the invention is being cultivated as a control group as experimental group with the conventional CIK cell for preparing
Cellular morphology figure after 9d.
Fig. 2 is the propagation quantity situation (× 10 after each group CIK cell culture 9d10)
Fig. 3 is cytoactive surface molecular mark expression after each group CIK cell culture 9d：CD3+CD56+ ratios (%)
Fig. 4 is cytoactive surface molecular mark expression after each group CIK cell culture 9d：CD4+CD25+ ratios (%)
Specific embodiment
The preparation of the CIK cell of the present invention of embodiment 1
A kind of preparation method of antitumor adoptive immunity competent cell CIK, it is characterised in that comprise the following steps：
1) collection and preparation of PMNC
1.1) 4 blood of cancer patients liquid samples are extracted.By venipuncture, liquaemin (40,000 U/L) anti-freezing is adopted respectively
Collection peripheral blood 80ml.The blood sample and physiological saline 1 that will be collected in superclean bench:1 volume ratio mixed diluting, lymph is thin
Born of the same parents separating liquid Ficoll-Paque PREMIUM separate PBMC, and lymphocyte separation medium is with dilution blood sample with 1:2 volume ratios are added
Centrifuge tube, at room temperature, 2500rpm centrifugations 20min；
1.2) the tunica albuginea layer of each blood sample of patient of collected after centrifugation, i.e. PBMC parts, add physiological saline adjustment volume extremely
50ml is simultaneously mixed, and PBMC is obtained after 1800rpm is centrifuged 5min, above step repeated washing cell 2 times.
2) preparation of CIK cell
2.1) adjustment cell density is 1 × 106/ ml, is inoculated in containing final concentration of 100ng/ml IL-6 and 100ng/ml
In the CIK cell basal medium of IL-21,37 DEG C, routinely it is incubated in 5%CO2 incubators；
2.2) after after 24h, cell is moved into the CIK cell basal medium containing final concentration of 1000U/ml IFN-γs
Middle cellar culture；
2.3) after after 24h to cultivating system in add final concentration of 500ng/ml people's thymus gland globulin, 500U/ml IL-2
And 10ng/ml IL-27；
2.4) hereafter every 3d supplement equivalent contain final concentration of 500U/ml IL-2 and 10ng/ml IL-27 CIK it is thin
Born of the same parents' basal medium, persistently cultivates 9d, can obtain high proliferation power, the antitumor adoptive immunity competent cell of cytotoxic activity high
CIK。
CIK cell basal medium is the modified form RPMI-1640 culture mediums of the autologous plasmas of 5-13ml/L containing final concentration.
The preparation of the conventional CIK cell of embodiment 2
The PBMC that will be obtained is washed twice, and is suspended in the modified form RPMI-1640 trainings of final concentration 5-13ml/L autologous plasmas
Support in base, adjustment cell density is 1 × 106/ ml, adds 1000U/ml IFN-γs, and final concentration of 100ng/ml is added after 24h
CD3mAb, the IL-2 of 1000U/ml, in 37 DEG C, be routinely incubated in 5%CO2 incubators.Persistently culture 9d collects cell and is used for
Subsequent detection.
Each Indexs measure of CIK cell is as follows：
The CIK cell morphologic detection of embodiment 3
By above-mentioned experimental group (embodiment 1) and control group (embodiment 2) when 9d is cultivated in carrying out cellular morphology under mirror
Observation, visible cellular control unit suspension growth in nutrient solution under mirror, cell is rounded, and quantity is few, occasionally has cell colony sample to give birth to
It is long；Experimental group cell quantity is significantly more than control group, and more in colony sample suspension growth, and cell colony group is more.
The CIK cell multiplication capacity of embodiment 4 and cell phenotype are detected
The experimental group (embodiment 1) and control group (embodiment 2) cell that 9d will be cultivated use flow cytomery each group
Cell quantity and the equimolecular expression of cell surface CD3, CD4, CD56, CD25.Despite the presence of individual difference, but experimental group
Ability of cell proliferation is obviously higher than control group；The ratio of CD3+CD56+ cells is above control group；Patient CD4+CD25+
Treg cell proportions are significantly less than control group.
Claims (3)
1. a kind of preparation method of antitumor adoptive immunity competent cell CIK, it is characterised in that comprise the following steps：
1) collection and preparation of PMNC (PBMC)：
1.1) by venipuncture, liquaemin (40,000 U/L) anti-freezing collection peripheral blood 50-100ml；In in superclean bench by blood
Liquid sample and physiological saline are with 1:1 volume ratio mixed diluting, human peripheral lymphocyte separating liquid (FICOLL configurations) is separated
PBMC, lymphocyte separation medium is with dilution blood sample with 1:2 volume ratios add centrifuge tube, at room temperature, 2000-2500rpm centrifugations 20-
30min；
1.2) collected after centrifugation tunica albuginea layer, i.e. PBMC parts；Volume is to 45-50ml and mixes to add physiological saline adjustment, in
1500-1800rpm obtains PBMC after 5-10min, above step repeated washing cell 2-3 times is centrifuged；
2) preparation of CIK cell：
2.1) adjustment cell density is 1 × 106/ml‐2×106/ ml, be inoculated in containing final concentration of 75-150ng/ml IL-6 and
In the CIK basal mediums of 75-150ng/ml IL-21,37 DEG C, routinely it is incubated in 5%CO2 incubators；
2.2) after 18-24h, cell is moved in the CIK basal mediums containing final concentration of 850-1200U/ml IFN-γs often
Rule culture；
2.3) after 18-24h, to adding final concentration of 350-650ng/ml people's thymus gland globulin 500-650U/ml in cultivating system
IL-2 and 5.5-13.5ng/ml IL-27；
2.4) hereafter contain final concentration 500-650U/ml IL-2 and 5.5-13.5ng/ml IL-27's every 3-4d supplement equivalent
In CIK basal mediums, 9-11d is persistently cultivated.
2. the preparation method of antitumor adoptive immunity competent cell CIK according to claim 1, it is characterised in that described
CIK cell basal medium is the modified form RPMI-1640 cultures of the autologous plasmas of 5-13ml/L containing final concentration in step (2.1)
Base.
3. the preparation method of antitumor adoptive immunity competent cell CIK according to claim 1, it is characterised in that including
Following steps：
1) collection and preparation of PMNC
1.1) 4 blood of cancer patients liquid samples are extracted.By venipuncture, liquaemin (40,000 U/L) anti-freezing gathers outer respectively
All blood 80ml.The blood sample and physiological saline 1 that will be collected in superclean bench:1 volume ratio mixed diluting, lymphocyte point
Chaotropic Ficoll-Paque PREMIUM separate PBMC, and lymphocyte separation medium is with dilution blood sample with 1:2 volume ratios add centrifugation
Pipe, at room temperature, 2500rpm centrifugations 20min；
1.2) the tunica albuginea layer of each blood sample of patient of collected after centrifugation, i.e. PBMC parts, add physiological saline to adjust volume to 50ml
And mix, obtain PBMC after 1800rpm is centrifuged 5min, above step repeated washing cell 2 times.
2) preparation of CIK cell
2.1) adjustment cell density is 1 × 106/ ml, is inoculated in containing final concentration of 100ng/ml IL-6 and 100ng/ml IL-21
CIK cell basal medium in, 37 DEG C, be routinely incubated in 5%CO2 incubators；
2.2) after after 24h, cell is moved in the CIK cell basal medium containing final concentration of 1000U/ml IFN-γs often
Rule culture；
2.3) after after 24h to cultivating system in add final concentration of 500ng/ml people's thymus gland globulin, 500U/ml IL-2 and
10ng/ml IL‐27；
2.4) hereafter the CIK cell base of final concentration of 500U/ml IL-2 and 10ng/ml IL-27 is contained every 3d supplement equivalent
Basal culture medium, persistently cultivates 9d, can obtain high proliferation power, the antitumor adoptive immunity competent cell CIK of cytotoxic activity high.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611204772.2A CN106754700A (en) | 2016-12-23 | 2016-12-23 | A kind of preparation method of antitumor adoptive immunity competent cell CIK |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611204772.2A CN106754700A (en) | 2016-12-23 | 2016-12-23 | A kind of preparation method of antitumor adoptive immunity competent cell CIK |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106754700A true CN106754700A (en) | 2017-05-31 |
Family
ID=58897827
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611204772.2A Pending CN106754700A (en) | 2016-12-23 | 2016-12-23 | A kind of preparation method of antitumor adoptive immunity competent cell CIK |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106754700A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106668067A (en) * | 2017-01-22 | 2017-05-17 | 暨南大学 | Antitumor combined medicine |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2511838A1 (en) * | 2002-12-31 | 2004-07-22 | Schering Corporation | Uses of mammalian cytokine; related reagents |
| CN103923880A (en) * | 2014-05-08 | 2014-07-16 | 成都百赛泰科生物科技有限公司 | Efficient multiplication CTL preparation method killing tumors in targeted mode |
| CN105713876A (en) * | 2016-03-29 | 2016-06-29 | 深圳爱生再生医学科技有限公司 | CIK (cytokine-induced killer) cell culture medium and application thereof |
| CN105886467A (en) * | 2014-08-27 | 2016-08-24 | 瑞安市普罗生物科技有限公司 | Preparation method of high-activity CIK cell |
| CN105969728A (en) * | 2016-05-21 | 2016-09-28 | 丁露露 | High-cytotoxic activity CIK cells and application in tumor cellular immunotherapy |
-
2016
- 2016-12-23 CN CN201611204772.2A patent/CN106754700A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2511838A1 (en) * | 2002-12-31 | 2004-07-22 | Schering Corporation | Uses of mammalian cytokine; related reagents |
| CN103923880A (en) * | 2014-05-08 | 2014-07-16 | 成都百赛泰科生物科技有限公司 | Efficient multiplication CTL preparation method killing tumors in targeted mode |
| CN105886467A (en) * | 2014-08-27 | 2016-08-24 | 瑞安市普罗生物科技有限公司 | Preparation method of high-activity CIK cell |
| CN105713876A (en) * | 2016-03-29 | 2016-06-29 | 深圳爱生再生医学科技有限公司 | CIK (cytokine-induced killer) cell culture medium and application thereof |
| CN105969728A (en) * | 2016-05-21 | 2016-09-28 | 丁露露 | High-cytotoxic activity CIK cells and application in tumor cellular immunotherapy |
Non-Patent Citations (3)
| Title |
|---|
| 孟明耀: ""改良 RPMI1640 培养基对 CIK 细胞增殖的影响"", 《中国生物制品学杂志》 * |
| 段玉青等: ""白细胞介素-27 促进 CIK 细胞的增殖及其对淋巴瘤细胞的杀伤能力"", 《中国肿瘤生物治疗杂志》 * |
| 陈伟等: ""Thymoglobulin 与 CD3 单克隆抗体在细胞因子诱导的杀伤细胞制备中的作用比较"", 《细胞与分子免疫学杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106668067A (en) * | 2017-01-22 | 2017-05-17 | 暨南大学 | Antitumor combined medicine |
| CN106668067B (en) * | 2017-01-22 | 2019-09-17 | 暨南大学 | A kind of antitumor combination medicine |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109294985B (en) | Culture medium system for NK cell in-vitro amplification and NK cell in-vitro amplification method | |
| CN101519646B (en) | CIK cell, as well as preparation method and cell preparation thereof | |
| CN103756963A (en) | Method used for in vitro proliferation of NK cells | |
| CN102321577B (en) | Preparation method of antitumor adoptive immune cells and prepared immune cells | |
| US11566227B2 (en) | Kit containing medium for culturing natural killer cells and method of effectively culturing natural killer cells using the same | |
| CN113046313A (en) | Composition and kit for efficiently inducing and amplifying human peripheral blood killer immune cells and culture method of immune cells | |
| CN105886469B (en) | CIK cell and its cultural method and application | |
| CN104480069A (en) | Method of carrying out isolated culture on immune cells by virtue of peripheral blood | |
| US10125351B2 (en) | Industrial preparations of natural killer (NK) cells and injections containing NK cells | |
| CN116875547A (en) | In-vitro amplification culture method for activating NK cells by utilizing exosomes | |
| CN105176926A (en) | Method for amplifying NK cells through in-vitro cultivation | |
| CN107974431B (en) | Rapid amplification method of natural killer cells | |
| CN105505871B (en) | A kind of effective amplification CIK and improve the method that its specificity kills tumor ability | |
| CN107502591B (en) | The iNKT methods for cell expansion and its application that a kind of concentration gradient rhIL-2 is relied on | |
| CN109010811B (en) | Immune preparation of broad-spectrum anti-tumor mouse model by using apoptotic body vaccine | |
| CN108642013A (en) | From being detached in Cord blood after CD34 candidate stem cells expand culture, induction prepares Dendritic Cells method to one kind on a large scale | |
| CN110747167B (en) | Preparation method and application of hemizygous BAK cell | |
| CN115896016B (en) | Culture composition and application thereof in culturing immune cells | |
| CN107502592A (en) | A kind of immunocyte serum free medium and preparation method thereof | |
| CN109957543A (en) | Utilize the method for Cord blood massive amplification Cord Blood Natural Killer Cells: Impact | |
| CN106754700A (en) | A kind of preparation method of antitumor adoptive immunity competent cell CIK | |
| CN105505872B (en) | Method and composition for sensitizing NK cells | |
| CN109535241B (en) | DC-CIK (dendritic cell-cytokine induced killer) co-culture cell, preparation method thereof, sensitizing antigen and application | |
| CN102212505B (en) | Immune killer cell, preparation method thereof, medicinal composition containing immune killer cell and set | |
| CN103937742B (en) | One activates CD4 simultaneously+& CD8+the cultural method of the immunocyte of T cell |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Xue Jing Inventor after: Guo Jing Inventor after: Chao Xiaodong Inventor after: Sun Bei Inventor before: Sun Bei Inventor before: Chao Xiaodong |