CN111849892B - In-vitro amplification method and application of glioma-derived Tumor Infiltrating Lymphocytes (TILs) - Google Patents
In-vitro amplification method and application of glioma-derived Tumor Infiltrating Lymphocytes (TILs) Download PDFInfo
- Publication number
- CN111849892B CN111849892B CN202010646970.4A CN202010646970A CN111849892B CN 111849892 B CN111849892 B CN 111849892B CN 202010646970 A CN202010646970 A CN 202010646970A CN 111849892 B CN111849892 B CN 111849892B
- Authority
- CN
- China
- Prior art keywords
- til
- glioma
- cells
- cell culture
- amplification method
- 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.)
- Active
Links
Images
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
- C12N5/0636—T lymphocytes
- C12N5/0638—Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/32—Amino acids
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/44—Thiols, e.g. mercaptoethanol
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/70—Undefined extracts
- C12N2500/80—Undefined extracts from animals
-
- 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]
- C12N2501/2315—Interleukin-15 (IL-15)
-
- 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
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
-
- 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
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
- C12N2509/10—Mechanical dissociation
Abstract
The embodiment of the invention discloses an in-vitro amplification method and application of glioma-derived Tumor Infiltrating Lymphocytes (TILs), belonging to the technical field of cell culture. The method comprises the following steps: pre-culturing TIL cells; and (3) culturing and expanding TIL cells. According to the invention, under the condition of not separating glioma infiltrating lymphocytes, TIL cells are directly induced and cultured and then are massively amplified, the operation procedure is simple, the time consumption is short, the amplification quantity and the cell activity of the TIL cells can be obviously improved, and the effect of killing glioma tumor cells is good. In the steps of TIL cell culture and amplification, the low-concentration cytokine composite nutrient medium is adopted, so that the requirement of TIL cell culture amplification is met, and the potential risk of cytokine storm in the subsequent treatment process is reduced.
Description
Technical Field
The embodiment of the invention relates to the technical field of cell culture, in particular to an in-vitro amplification method and application of glioma-derived Tumor Infiltrating Lymphocytes (TILs).
Background
Glioma is the most common primary intracranial tumor originated from brain neuroepithelium, is difficult to cure radically and is easy to relapse. The main treatment methods at present are surgical treatment, radiotherapy, chemotherapy and the like. The 5-year survival of high grade gliomas, especially glioblastoma, is less than 10%.
Tumor Infiltrating Lymphocytes (TILs) are lymphocytes isolated from tumor tissue. The existing TIL cell culture technology is to separate tumor infiltrating lymphocytes from tumor tissues by using a mechanical treatment and/or enzyme digestion method, and add high-dose interleukin-2 (IL-2, 6000U/mL) to carry out in-vitro culture. Studies have shown that IL-2-activated TILs have higher tumoricidal activity and better targeting than LAK from autologous Peripheral Blood Mononuclear Cells (PBMCs).
TIL cells have been used for more than 20 years as a specific immune cell for tumor therapy and have been studied in a variety of tumors. The currently used TIL in vitro amplification technology is to separate TIL cells from tumor tissues and then perform TIL induction culture, and the TIL cells cultured in vitro have been proved to have the anti-tumor effect. However, the existing method for expanding TIL cells needs to separate TIL cells first, which has the defects of slow expansion speed, small number of obtained lymphocytes and the like, and in order to obtain more effector cells quickly, the dosage of cytokines (such as IL-2) is higher, and the patients are easy to generate factor storm in the treatment process, so that the method is not beneficial to treatment and even generates life risk to the patients.
The related report of glioma TIL is not found in the prior art, so that the development of a novel method for efficiently amplifying glioma TIL in vitro is of great significance.
Disclosure of Invention
Therefore, the embodiment of the invention provides an in vitro amplification method of glioma-derived Tumor Infiltrating Lymphocytes (TILs) and application thereof, so as to solve the problems of complicated operation procedures, long time consumption, low activity and high IL-2 usage amount of the existing TIL cell amplification method.
In order to achieve the above object, the embodiments of the present invention provide the following technical solutions:
according to a first aspect of embodiments of the present invention, there is provided a method for in vitro expansion of glioma-derived Tumor Infiltrating Lymphocytes (TILs), the method comprising the steps of:
(1) Pre-culture of TIL cells
The glioma tissue is washed by PBS and cut into 1-3mm 3 The size of the tissue mass was adjusted to 37 ℃ and 5% CO using the preculture solution 2 Culturing in an incubator to fully release PBMC in the tissue mass capillaries;
(2) Culture and expansion of TIL cells
The glioma tissue obtained in step (1) is incubated in TIL cell culture medium I at 37 deg.C and 5% CO 2 Culturing in an incubator, changing the culture solution once every 2-3 days, adding a TIL cell culture medium II after culturing for 6-8 days, continuing culturing for 6-8 days, centrifuging at 800-1000rpm/min for 5-8min, removing supernatant, resuspending and washing cells by using pre-culture solution, separating, purifying and collecting TIL cells for further experiment.
Further, the composition of the pre-culture solution is as follows: addition of 1% P/S (penicillin/streptomycin) to RMPI 1640 medium.
Further, the composition of the TIL cell culture medium i is as follows: adding 5% of human A/B serum, 1% of human platelet lysate, 1% of P/S, 2mM L-glutamine, 1 XMEM-Eagle, 1mM Sodium pyrolate, 10mM HEPES, 1 XBeta-mercaptoethanol, 500U/mL IL-2, 50U/mL IL-15, 50U/mL IL-21 to the GMP DC medium.
Further, the composition of the TIL cell culture medium ii was as follows: adding 10% of human A/B serum, 1% of human platelet lysate, 1% of P/S, 2mM L-glutamine, 1 XMEM-Eagle, 1mM Sodium pyrolate, 10mM HEPES, 1 XBeta-mercaptoethanol, 1000U/mL IL-2, 100U/mL IL-15, 100U/mL IL-21 to the GMP DC medium.
Further, in step (1), glioma tissues were seeded in 100mm cell culture dishes.
Further, in the step (1), the culture time is 30min.
Further, in step (2), glioma tissues were seeded in 24-well plates, 2 pieces/well.
According to a second aspect of embodiments of the present invention, there is provided the use of Tumour Infiltrating Lymphocytes (TILs) obtained by the in vitro expansion method described above in the treatment of glioma.
The embodiment of the invention has the following advantages:
1. according to the invention, under the condition that glioma infiltrating lymphocytes are not separated, a great amount of TIL cells are expanded after TIL cells are directly induced and cultured, the operation procedure is simple, the time consumption is short, the expansion amount and the cell activity of the TIL cells can be obviously improved, and the effect of killing glioma tumor cells is good.
2. In the steps of TIL cell culture and amplification, the low-concentration cytokine composite nutrient medium is adopted, so that the requirement of TIL cell culture amplification is met, and the potential risk of cytokine storm in the subsequent treatment process is reduced.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.
FIG. 1 is a culture diagram of glioma primary tumor cells of the present invention;
FIG. 2 is a diagrammatic representation of a TIL cell culture of the present invention;
FIG. 3 shows the flow cytometry results of TILs of the present invention (including the detection of CD45, CD3, CD4, CD8 content);
FIG. 4 shows the flow cytometry results of TILs of the invention (including the detection of CD19, CD127, CD4, CD8 content);
FIG. 5 is a schematic diagram of the process of killing glioma tumor tissue cells by TIL cells of the present invention.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Culture of glioma tumor tissue cells
Taking part of the glioma tumor tissue, grinding, digesting, removing red blood cells, centrifuging at 800rpm/min for 5min to collect cells, discarding the supernatant, and subjecting the supernatant to 5% CO at 37 deg.C 2 And culturing the cells in the incubator. Wherein the culture solution comprises the following components: adding 10% of FBS, 1% of P/S (penicillin/streptomycin), 2mM L-glutamine, 1 XMEM-Eagle, 1mM Sodium pyruvate pyroltate, 10mM HEPES, based on DMEM/F12 medium. After 1-3 generations of culture, the cells are digested, centrifuged at 800rpm/min for 5min to collect the cells, resuspended and washed by adding culture medium, and then stored or subjected to the next experiment. The culture results of primary tumor cells of glioma are shown in figure 1.
Example 2
Method for in vitro expansion of glioma infiltrating lymphocytes (TILs)
(1) Pre-culture of TIL cells
The collection tube containing 1% P/S of RMPI 1640 medium was placed on ice, and glioma tissues collected in the operating room were quickly placed in the collection tube, which is advantageous for preserving the activity of the tissues during transport.
Taking out glioma tissue, washing with 1 × PBS for 3 times, and cutting into 1-3mm 3 Inoculating the tissue mass into a 100mm cell culture dish containing 1% P/S RMPI 1640 medium, placing in 5% CO 2 And culturing in an incubator at 37 ℃ for 30min to fully release the PBMC in the capillary vessel of the tissue block.
(2) Culture and expansion of TIL cells
The glioma tissues obtained in step (1) were inoculated in 24-well plates containing TIL cell culture medium I, 2 blocks/well, incubated at 37 ℃ and 5% CO 2 Culturing in an incubator, changing the culture medium once every 2 days, culturing for 8 days, adding TIL cell culture medium II, and continuing culturingFor 8 days, centrifuge at 1000rpm/min for 5min, discard the supernatant, resuspend and wash the cells with media.
The composition of the TIL cell culture medium I is as follows: adding 5% of human A/B serum, 1% of human platelet lysate, 1% of P/S, 2mM L-glutamine, 1 XMEM-Eagle, 1mM Sodiumpyrolate, 10mM HEPES, 1 XBeta-mercaptoethanol, 500U/mL IL-2, 50U/mL IL-15, 50U/mL IL-21 to the GMP DC medium.
The composition of TIL cell culture medium ii was as follows: adding 10% of humann A/B serum, 1% of human platelet lysate, 1% of P/S, 2mM L-glutamine, 1 XMEM-Eagle, 1mM Sodiumpyrolate, 10mM HEPES, 1X β -mercaptoethanol, 1000U/mLIL-2, 100U/mL IL-15, 100U/mL IL-21 to the GMP DC medium.
The results of TIL cell culture are shown in fig. 2.
Example 3
TIL cell composition analysis
Flow cytometry analysis of CD3 in TIL cells released from glioma tissue blocks obtained by the amplification method of example 2 + -T，CD4 + -T，CD8 + -T，CD19 + -T, and CD127 + T, see FIG. 3, FIG. 4 and Table 1.
TABLE 1TIL cell component content and expansion (including CD4 and CD 8)
The results showed that CD8 was present in expanded culture of TIL cells in vitro + The proportion of T cells is at a steady high level, and CD8 + The proportion of T cells increases continuously during the slow expansion phase and at a steady high level during the fast expansion phase.
Example 4
In vitro killing activity of TIL cells against glioma tumor tissue cells
TIL cells (referred to as effector cells) cultured in example 2 were collected and cultured in mixed culture with glioma tumor tissue cells (referred to as target cells) cultured in example 1, with the ratio of effector cells to target cells set at 1. The killing effect of the TIL cells obtained in example 2 on glioma tumor tissue cells was tested by taking pictures at different time periods (after mixed culture, the shooting test was carried out on days 2,4 and 6 respectively), and the results are shown in FIG. 5.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (6)
1. A method for the in vitro expansion of Tumor Infiltrating Lymphocytes (TILs) of glioma origin, comprising the steps of:
(1) Pre-culture of TIL cells
The glioma tissue is washed by PBS and cut into 1-3mm 3 The size of the tissue mass was adjusted to 37 ℃ and 5% CO using the preculture solution 2 Culturing in an incubator to fully release PBMC in the tissue mass capillaries;
(2) Culture and expansion of TIL cells
The glioma tissue obtained in step (1) is treated with TIL cell culture medium I at 37 ℃ and 5% CO 2 Culturing in an incubator, changing the culture solution once every 2 to 3 days, culturing for 6 to 8 days, adding a TIL cell culture medium II, continuously culturing for 6 to 8 days, centrifuging at 800 to 1000rpm for 5 to 8min, discarding supernatant, resuspending and washing cells by using the preculture solution, separating, purifying and collecting TIL cells for next experiment;
the composition of the TIL cell culture medium I is as follows: adding 5% human A/B serum, 1% human platelet lysate, 1% P/S, 2mM L-glutamine, 1 XMEM-Eagle, 1mM Sodium pyrolate, 10mM HEPES, 1 XBeta-mercaptoethanol, 500U/mL IL-2, 50U/mL IL-15, 50U/mL IL-21 to the GMP DC medium;
the composition of the TIL cell culture medium II is as follows: adding 10% of human A/B serum, 1% of human platelet lysate, 1% of P/S, 2mM L-glutamine, 1 XMEM-Eagle, 1mM Sodium pyrolate, 10mM HEPES, 1 XBeta-mercaptoethanol, 1000U/mL IL-2, 100U/mL IL-15, 100U/mL IL-21 to the GMP DC medium.
2. The in vitro amplification method of claim 1, wherein the composition of the pre-culture fluid is as follows: adding 1% P/S to RMPI 1640 culture medium.
3. The in vitro amplification method of claim 1, wherein in step (1), the glioma tissue is seeded in a 100mm cell culture dish.
4. The in vitro amplification method of claim 1, wherein the culture time in step (1) is 30min.
5. The in vitro amplification method of claim 1, wherein in step (2), the glioma tissue is seeded in 24-well plates at 2 blocks/well.
6. Use of Tumor Infiltrating Lymphocytes (TILs) obtained by the in vitro amplification method of claim 1 in the preparation of a medicament for treating glioma.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010646970.4A CN111849892B (en) | 2020-07-07 | 2020-07-07 | In-vitro amplification method and application of glioma-derived Tumor Infiltrating Lymphocytes (TILs) |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010646970.4A CN111849892B (en) | 2020-07-07 | 2020-07-07 | In-vitro amplification method and application of glioma-derived Tumor Infiltrating Lymphocytes (TILs) |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111849892A CN111849892A (en) | 2020-10-30 |
| CN111849892B true CN111849892B (en) | 2023-02-03 |
Family
ID=73152964
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010646970.4A Active CN111849892B (en) | 2020-07-07 | 2020-07-07 | In-vitro amplification method and application of glioma-derived Tumor Infiltrating Lymphocytes (TILs) |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111849892B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109536444B (en) * | 2018-12-11 | 2022-06-28 | 吉林省拓华生物科技有限公司 | Separation induction method suitable for malignant solid tumor infiltrating T lymphocytes |
| CN113215099B (en) * | 2021-04-28 | 2022-11-22 | 广东康盾创新产业集团股份公司 | TIL cell amplification culture medium and use method thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103396991A (en) * | 2013-05-02 | 2013-11-20 | 陈晚华 | Method for rapidly and efficiently amplifying tumor infiltrating lymphocytes |
| CN105713878A (en) * | 2015-12-21 | 2016-06-29 | 杭州特马赛生物技术有限公司 | Method for in-vitro expansion of CD8<+>T cells |
| CN107043749A (en) * | 2017-04-21 | 2017-08-15 | 北京奥康华医学检验所有限公司 | A kind of separant induction method of tumor-infiltrated T lymphocytes |
| CN107384867A (en) * | 2017-08-04 | 2017-11-24 | 北京世纪劲得生物技术有限公司 | A kind of tumor tissues til cell preparation method and special culture media |
| CN109536444A (en) * | 2018-12-11 | 2019-03-29 | 吉林省拓华生物科技有限公司 | A kind of separant induction method suitable for the tumor-infiltrated T lymphocyte of malignant solid tumor |
| EP3517600A1 (en) * | 2018-01-24 | 2019-07-31 | Zellwerk GmbH | Meander bioreactor and method for the isolation and proliferation of cells from parts from tumours, metastases and other tissues |
| CN110643574A (en) * | 2019-10-30 | 2020-01-03 | 西南医科大学 | Preparation method of feeder cells for rapid culture of tumor infiltrating lymphocytes |
| CN110713978A (en) * | 2019-11-16 | 2020-01-21 | 福建省肿瘤医院（福建省肿瘤研究所、福建省癌症防治中心） | Separation method of tumor antigen specific tumor infiltrating T cells |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20170044496A1 (en) * | 2014-04-10 | 2017-02-16 | H. Lee Moffitt Cancer Center And Research Institute, Inc. | Enhanced Expansion of Tumor-Infiltrating Lymphocytes for Adoptive Cell Therapy |
-
2020
- 2020-07-07 CN CN202010646970.4A patent/CN111849892B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103396991A (en) * | 2013-05-02 | 2013-11-20 | 陈晚华 | Method for rapidly and efficiently amplifying tumor infiltrating lymphocytes |
| CN105713878A (en) * | 2015-12-21 | 2016-06-29 | 杭州特马赛生物技术有限公司 | Method for in-vitro expansion of CD8<+>T cells |
| CN107043749A (en) * | 2017-04-21 | 2017-08-15 | 北京奥康华医学检验所有限公司 | A kind of separant induction method of tumor-infiltrated T lymphocytes |
| CN107384867A (en) * | 2017-08-04 | 2017-11-24 | 北京世纪劲得生物技术有限公司 | A kind of tumor tissues til cell preparation method and special culture media |
| EP3517600A1 (en) * | 2018-01-24 | 2019-07-31 | Zellwerk GmbH | Meander bioreactor and method for the isolation and proliferation of cells from parts from tumours, metastases and other tissues |
| CN109536444A (en) * | 2018-12-11 | 2019-03-29 | 吉林省拓华生物科技有限公司 | A kind of separant induction method suitable for the tumor-infiltrated T lymphocyte of malignant solid tumor |
| CN110643574A (en) * | 2019-10-30 | 2020-01-03 | 西南医科大学 | Preparation method of feeder cells for rapid culture of tumor infiltrating lymphocytes |
| CN110713978A (en) * | 2019-11-16 | 2020-01-21 | 福建省肿瘤医院（福建省肿瘤研究所、福建省癌症防治中心） | Separation method of tumor antigen specific tumor infiltrating T cells |
Non-Patent Citations (2)
| Title |
|---|
| Tumor-infiltrating lymphocytes (TILs) from patients with glioma;Zhenjiang Liu et al.;《OncoImmunology》;20170208;第6卷(第2期);第1-9页 * |
| 人脑胶质瘤浸润淋巴细胞的制备及测活;梁兵等;《济宁医学院学报》;20001231;第23卷(第4期);第50页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111849892A (en) | 2020-10-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109294985B (en) | Culture medium system for NK cell in-vitro amplification and NK cell in-vitro amplification method | |
| CN103756963A (en) | Method used for in vitro proliferation of NK cells | |
| CN102154206B (en) | Preparation method of high-purity, high-multiplication capacity and high-cytotoxin activity CIK (cytokine induced kill) cell | |
| CN111849892B (en) | In-vitro amplification method and application of glioma-derived Tumor Infiltrating Lymphocytes (TILs) | |
| CN102321577B (en) | Preparation method of antitumor adoptive immune cells and prepared immune cells | |
| CN108588022B (en) | Method for enriching human CD4+ and CD8+ TCM cells through in vitro culture | |
| CN108676775B (en) | Method for amplifying cord blood NK in vitro | |
| CN110643574A (en) | Preparation method of feeder cells for rapid culture of tumor infiltrating lymphocytes | |
| CN115678846B (en) | Tumor specific gamma delta T cell and preparation method thereof | |
| WO2022143785A1 (en) | Methods for preparing tumor-infiltrating lymphocytes | |
| Najar et al. | Cytokinome of adult-derived human liver stem/progenitor cells: immunological and inflammatory features | |
| CN103372029A (en) | NK (Natural Killer) cell new technology for treating tumor | |
| CN107502589A (en) | A kind of tumor infiltrating lymphocyte and mononuclearcell co-culture method | |
| CN117089518A (en) | Exosome preparation method, product and application thereof | |
| CN106047809A (en) | Method for combining with ligand TLR7 to simultaneously amplify human CIK/NK cells | |
| CN111286486A (en) | Method for amplifying large amount of lymphocyte infiltrated by malignant tumor ascites in vitro | |
| CN101429495A (en) | Cultivation method for human peripheral blood source hemopoietic stem cell | |
| CN101429496A (en) | Culture medium for human peripheral blood source hemopoietic stem cell | |
| CN103710308B (en) | Muramyl dipeptide is utilized to induce the method for DC-CIK | |
| CN114032211A (en) | Application of trametes acetyl acid in CIK cell in-vitro amplification | |
| CN108004213B (en) | Method and kit for rapid amplification of CIK cells | |
| CN113005085A (en) | Novel method for culturing and in-vitro amplifying primary liver cancer tumor infiltrating lymphocytes | |
| CN108504636A (en) | A kind of efficient NK cell culture processes | |
| CN116179486B (en) | Preparation method of tumor-infiltrating lymphocytes | |
| CN113265376B (en) | Activation method of TIL (dendritic cells) |
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: Wu Dinglan Inventor after: Yang Yinggui Inventor after: Li Xin Inventor after: Mao Jie Inventor after: Zhou Jiayi Inventor after: Ding Tengteng Inventor before: Wu Dinglan |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |