CN102793912A - Dendritic cell (DC) tumor vaccine and preparation method thereof - Google Patents
Dendritic cell (DC) tumor vaccine and preparation method thereof Download PDFInfo
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Abstract
The invention relates to dendritic cell (DC) tumor vaccine in which a multi-medicine resistance tumor stem cell antigen composition is loaded. The antigen composition loaded in the DC tumor vaccine derives from tumor tissues, and is mainly used for tumor stem cells; and specifically, the tumor tissues are subjected to tumor stem cell primary separation and culture so as to obtain the tumor stem cells, the tumor stem cells are screened by a flow cytometry so as to obtain tumor stem cells with multi-medicine resistance, and finally full cell antigens are prepared.
Description
Technical field
The present invention relates to a kind of biovaccine and preparation method thereof, particularly relate to a kind of tumor vaccine and preparation method thereof.
Background technology
Conventional tumor therapeuticing method comprises operative treatment, radiation cure and chemotherapy etc.But existing tumor therapeuticing method often can not solve the problem of tumor recurrence.Main cause is to constitute in the cell of tumor to contain tumor stem cell, and it has the biological characteristics of stem cell, i.e. self renewal, pluripotency differentiation, height endurability are promptly to the high drug-resistance of chemotherapeutics.Therefore tumor stem cell is the real arch-criminal that tumor takes place and develops.
BMDC (DC, Dendritic Cell) is the strongest full-time antigen presenting cell of function in the human body, has the function of activation primary tape T cell.Generally DC is equivalent to the courier, and it is the T cell that antigenic information is passed to the effect person that can bring into play immunological effect and killing ability.Dendritic cell tumor vaccine (DC tumor vaccine) is the functional BMDC of carrying tumor antigen information, can activate specific T-cells and produce immunne response, produces GVT and immunological memory function.
The advantage of DC tumor vaccine is induced DC with tumor antigen and is made it that tumor antigen information is passed to the T cell, and the T cell can be re-recognized and killing tumor cell.Behind the external impact sensitization of multi-form tumor antigen DC, can produce stronger antineoplastic immune, and can suppress tumor growth and transfer.Can in patient's body, bring out immunological memory, make to obtain secular anti-pain effect in patient's body, prevent the recurrence of tumor.
Cytokine induced kill cell (CIK; Cytokine induced killer cell) be a kind of novel immunologically competent cell; It is a group foreign cell that nucleus is obtained after a period of time in external use various kinds of cell factor co-cultivation; Because this kind cell expresses CD3 and two kinds of membrane protein molecules of CD56 simultaneously, thus the NK cell appearance T lymphocyte that is otherwise known as, the CIK cell have growth rate fast, kill tumor activity high, kill the tumor spectrum and extensively reach the restricted advantage of tumor extremely of non-MHC.
CIK cell, the clinical treatment that can be applicable to multiple cancer different phase of DC tumor vaccine, as: pulmonary carcinoma (small cell lung cancer, scale cancer, adenocarcinoma etc.) and metastatic carcinoma thereof.Digestive system: esophageal carcinoma, gastric cancer, hepatocarcinoma, cancer of pancreas, knot, rectal cancer and metastatic carcinoma thereof.Urinary system: renal carcinoma, adrenal carcinoma, and metastatic carcinoma.Gynecological tumor: uterus carcinoma, ovarian cancer and metastatic carcinoma thereof.Blood system: acute and chronic leukemia, lymphoma (except t cell lymphoma), and metastatic carcinoma.Other tumors: malignant melanoma, nasopharyngeal carcinoma, breast carcinoma, carcinoma of prostate, carcinoma of tongue or the like.Use so far, do not see great clinical adverse report such as lesions of liver and kidney, bone marrow depression, the property crossed a parainfluenza appearance symptom can appear in few part patient, and like heating, headache, joint aches gets final product by conventional treatment, but spontaneous remission mostly.Clinical research shows that DC and CIK co-culture of cells can strengthen the quantity and the killing activity of CIK cell greatly, improves the clinical efficacy of CIK cell therapy.Utilize the DC cell and the CIK cell of load tumor antigen to hatch altogether, can promote propagation and the immunologic function of cell and DC simultaneously, thereby the wide spectrum of performance CIK cell kills the double effect of tumor and the activated specificity antineoplastic of DC.
DC tumor vaccine of the prior art all is to be target with the common cell that swells and ache, and promptly obtains antigen through the common cell that swells and ache of cracking, and with this antigen load to patient's BMDC.The BMDC of the target tumor cell that so obtains is used to treat cancer, has tumor high problem of drug resistance increase, recurrence and the rate of transform after chemotherapy.Reason is that the tumor stem cell in the tumor is not all killed, and causes the revivable growth of cancerous tissue, even the tumor stem cell of only a few also drives the formation of tumor.
Summary of the invention
The technical problem that the present invention solved provides a kind of new dendritic cell tumor vaccine (DC tumor vaccine); It is to be target with the tumor stem cell; Load the antigen compsn that obtains through the cracking tumor stem cell; At direct targeting kill tumor stem cell of when treatment, the cure rate of raising tumor disease.The present invention simultaneously also obtains antigen compsn through the tumor stem cell that screening has a multi-drug resistant, makes it be applicable to all kinds of patients.
Another object of the present invention provides the method for preparing of above-mentioned dendritic cell tumor vaccine.
The technical scheme that the present invention adopts is: a kind of dendritic cell tumor vaccine, its load have multi-drug resistant tumor stem cell antigen compsn.
The antigen compsn of load comes from tumor tissues itself in the DC tumor vaccine provided by the invention; Be primarily aimed in tumor stem cell; Specifically be with obtaining tumor stem cell after the former generation separation of tumor tissues process tumor stem cell, the cultivation; Screen the tumor stem cell with multi-drug resistant through flow cytometer again, the full cellular antigens of process prepare and obtain at last.
The method for preparing of dendritic cell tumor vaccine of the present invention may further comprise the steps:
(1) tumor tissues is carried out former generation separates and cultivation obtains tumor stem cell;
(2) adopt the flow cytometer screening to have the tumor stem cell of multi-drug resistant;
(3) tumor stem cell that step (2) is obtained prepares full cellular antigens;
(4) isolated dendritic cell and cultivation;
(5) the full cellular antigens that step (3) obtained and the BMDC of step (4) are cultivated altogether and obtain the dendritic cell tumor vaccine that load has multi-drug resistant tumor stem cell antigen compsn.
Owing to utilize the DC cell of load tumor antigen and CIK cell to hatch altogether, can promote the propagation and the immunologic function of CIK cell and DC cell simultaneously, so the method for preparing of dendritic cell tumor vaccine of the present invention can also be following steps:
(1) tumor tissues is carried out former generation separates and cultivation obtains tumor stem cell;
(2) adopt the flow cytometer screening to have the tumor stem cell of multi-drug resistant;
(3) tumor stem cell that step (2) is obtained prepares full cellular antigens;
(4) isolated dendritic cell (DC cell) and cytokine induced kill cell (CIK cell) and cultivation;
(5) the full cellular antigens that step (3) obtained and the BMDC of step (4) and cytokine induced kill cell are cultivated altogether and are obtained the dendritic cell tumor vaccine that load has multi-drug resistant tumor stem cell antigen compsn.
The tumor tissues that the present invention adopts is isolated tumor tissue in vitro in patient's body; Be the garbage in the clinical medicine; Can be after exsomatizing in the suitably medium-term and long-term preservation of environment; And in the former generation of tumor stem cell, separate and culture technique has been the ten minutes mature technique, but stem cell long preservation and increment fast in external environment, but clinical practice.And the method for preparing of the screening of chemical sproof tumor stem cell and full cellular antigens also has been known technology.
DC cell of the present invention or CIK cell come from peripheral blood or Cord blood, are excellent to come from Cord blood.Peripheral blood and Cord blood can obtain through the mechanism of special storage, like blood bank or umbilical cord blood bank, also can with getting with usefulness, be suitable for clinical practice in the suitably medium-term and long-term preservation of environment.And the separation of DC cell or CIK cell and culture technique also are known technology.
The method for preparing of dendritic cell tumor vaccine of the present invention can adopt various tumor tissue in vitro to prepare, like pulmonary carcinoma esophageal carcinoma, gastric cancer, hepatocarcinoma, cancer of pancreas, knot, rectal cancer, renal carcinoma, uterus carcinoma, ovarian cancer tissue or the like.Preferred breast tumor tissues.Wherein the mechanism of the screening of multi-drug resistant tumor stem cell is based on the mechanism of drug resistance of tumor chemotherapy; Mostly be because the expression of multidrug resistant gene (MDR1) and product P-glycoprotein (Pgp) thereof is increased; Therefore this method is based on the ultimate principle of antigen antibody reaction; Adopt Flow Cytometry; The tumor stem cell that cultivation is obtained carries out the detection of multidrug resistant gene (MDR1) and product P-glycoprotein (Pgp) thereof, filters out these two surface markers and is all positive cells, is the tumor stem cell of multi-drug resistant.
Conventional with common tumor stem cell be the cancer immunotherapy method of target still exist behind the radiation and chemotherapy tumor toleration especially drug resistance increase the problem that the relapse rate and the rate of transform are high.The present invention is directed to tumor stem cell, use the antigen load for preparing to BMDC, and come the specific killing tumor stem cell with the gained dendritic cell vaccine with multi-drug resistant.Dendritic cell vaccine has following advantage with respect to the vaccine of protein, molecular level: 1. preparation is simple; 2. include multiple t cell epitope, can guarantee the comprehensive of immunity and strong effect property; 3. meet heterogeneous tumor individualized treatment scheme.
Description of drawings
Fig. 1 is that the DC-CIK cell vaccine kills the tumor experimental result picture;
Fig. 2 is that the DC cell vaccine kills the tumor experimental result picture.
The specific embodiment
For further specifying the present invention, specify in conjunction with following examples:
Present embodiment is a source of human stem cell with preferred breast tumor tissues; The DC tumor vaccine for preparing is used to prepare the medicine of treatment breast tumor disease; But the present invention is not limited only to breast tumor; It can be a source of human stem cell with any other tumor tissues, and the similar or those skilled in the art of preparation process and therapeutic effect and present embodiment can obtain through the limited number of time test.
One, the isolated culture method of breast carcinoma stem cell
Tumor tissues with containing antibiotic normal saline flushing, is shredded with the ophthalmologic operation shears, 37 ℃ of enzymic digestions 2 hours, and filter with the cell sieve, cell suspension is used Stemedia
TMWIT-I
TMCulture Medium (stemgent) cultivates.
This culture medium is aimed at the epithelial special culture media of breast carcinoma, need not other cell growth factor and makes cell remain the form of chamber epithelium shape cell, and cell can gradually change into the shape of similar adenocarcinoma cell from last epithelial cell form.
Change Stemedia again into
TMWIT-P
TMCulture Medium (stemgent) culture medium culturing changed liquid once in three days, went down to posterity once in 5 days.
Two, the evaluation of breast carcinoma stem cell
Use flow cytometer that the breast carcinoma stem cell surface marker of being cultivated is cultivated, phenotype is CD45-CD31-TER119-(Lin-) CD29+CD24+, meets the cell phenotype of breast carcinoma stem cell fully.
The screening of multi-drug resistant breast carcinoma stem cell:
Because tumor stem cell possibly have autoprotection mechanism to poisonous substance (like chemotherapeutics), and chemotherapeutics is had more drug resistance.Therefore isolate tumor stem cell, and it is carried out the treatment of targeting, for the clinical treatment of tumor provides directive function with multi-drug resistant.
Learn in the present document that the mechanism of drug resistance of tumor chemotherapy is because the expression of multidrug resistant gene (MDR1) and product P-glycoprotein (Pgp) thereof is increased.Therefore the present invention is based on the ultimate principle of antigen antibody reaction; Adopt Flow Cytometry; The breast carcinoma stem cell that cultivation is obtained carries out the detection of multidrug resistant gene (MDR1) and product P-glycoprotein (Pgp) thereof; Filter out these two surface markers and be all positive cells, be the multi-drug resistant breast carcinoma stem cell.The selection markers of the multi-drug resistant of other tumor stem cells also is these two kinds, and screening technique is similar.
Three, the method for preparing of tumor stem cell antigen compsn
With normal saline washing and resuspended stem cell;
Tumor stem cell heat shock: in 39 ℃ of water-baths, hatched stem cell 2 hours;
The tumor stem cell multigelation: the full cell of breast carcinoma stem cell with multi-drug resistant experiment that uses screening to obtain carries out multigelation, makes lysis, thereby discharges full cellular antigens, is removed to select tumor antigen and it is processed, presents by DC.Refrigerated temperature is chosen as subzero 196 degree, and the temperature of thawing is chosen as room temperature.
Because through after the heat shock, tumor stem cell of the present invention can produce heat shock protein, still has said heat shock protein after the cracking, the dendritic cell vaccination that the having of heat shock protein helps after the load is replied.
Cracking is removed cell debris excessive in the lysate through centrifugal and filtering step after accomplishing usually.Centrifugal 600rpm, 1 minute, supernatant was used the 0.45um membrane filtration.
Four, preparation DC
Derive from the Cord blood or the peripheral blood of blood bank, obtain White Blood Cells Concentrate with lymphocyte separation medium, normal saline washing obtain fresh mononuclear cell.
DC induces: be laid on behind the mononuclear cell with the resuspended prepared fresh of RPM1640 culture medium in the 6 porocyte culture plates; Be positioned over 37 ℃, behind the 90min, flush away is attached cell not in the 50ml/L CO2 incubator; In culture plate, add and contain 100ug/LCM-CSF; The 50ug/LRPM1640 complete medium changed liquid on the 3rd day, the 5th day results iDC.
Antigen load DC: with the resuspended imDC of RPM1640 complete medium, add the cellular antigens of aforementioned preparation, 37 ℃, 50ml/L CO2 incubator co-cultivation 48 hours, results DC cell.Carrying out cell technology and phenotype detects.
Five, load multi-drug resistant breast carcinoma stem cell vaccine is to the research of the lethal effect of breast carcinoma stem cell
Adopt CCK-8 to detect and cultivate altogether with CIK and the DC and the CIK that do not have an antigen load cultivates altogether, single culture CIK is to the killing activity from the former foster tumor stem cell of being commissioned to train of body, be confirmatory experiment from the DC of body/alloantigen load.
The DC vaccine of the antigen compsn preparation of multi-drug resistant breast carcinoma stem cell is tested the lethal effect of tumor stem cell:
1, the external evoked amplification of DC, CIK, antigen load and phenotype detect
Separate the external evoked preparation of Cord blood mononuclearcell DC, CIK cell.Collect DC, CIK cell in 8d days, carry out common cultivation, be divided into no antigen DC group; The full cellular antigens submission of autologous tumor DC group; Allogeneic tumor cell lysate antigen presentation DC group with 10: 1 ratios and the CIK Mixed culture that is cultured to 8d, is carried out the target cell killing experiments after three days.Flow cytometer detects the expression of DC surface-ripe sign CD83, CD80, CD86, HLA-DR, the expression of CIK cell detection CD3, CD4, CD8, CD56.
This product detects the tumor cytotoxicity effect: use the CCK-8 method and detect CIK, CIK-DC, CIK-DC-tumor stem cell antigen, respectively to former be commissioned to train support and the multi-drug resistant tumor stem cell of screening kill tumor activity.
CCK-8 reagent can be used for easy and cell proliferation and oxicity analysis accurately.Its ultimate principle is: [its chemical name is: 2-(2-methoxyl group-4-nitrobenzophenone)-3-(4-nitrobenzophenone)-5-(2 to contain WST-8 in this reagent; 4-disulfonic acid benzene)-and 2H-tetrazolium list sodium salt], it is reduced to the yellow methyl dyestuff (Formazan dye) with high water soluble by the dehydrogenase in the cell mitochondrial under the effect of electron carrier 1-methoxyl group-5-toluphenazine DMS (1-Methoxy PMS).The quantity of the methyl compound that generates is directly proportional with the quantity of living cells, therefore can utilize this characteristic directly to carry out cell proliferation and oxicity analysis.
2, experimental result
According to CCK-8 test kit (Cell Counting Kit-8; The green skies, Shanghai Bioisystech Co., Ltd) description is set up matched group; This test is divided into four groups, and one group is the CIK group, and one group is CIK-DC Mixed culture group; One group is the CIK-DC group of tumor stem cell antigen load, and one group is the matched group that has only tumor stem cell.Use CCK-8 test kit detection specificity target cell kill rate.Measure the absorbance of 492nm, at last according to formula [(test hole-SR effector lymphocyte-SR target cell)/(MR target cell-SR target cell)] * 100%.Multiple 3 times of experiment gross weight carries out analyzing and processing to the result, respectively organizes the difference of cell killing rate.
CCK-8 reagent E LISA method testing result calculates the result and sees table 1 and Fig. 1:
Table 1
| Sequence number | Imitate the target ratio | The DC-CIK of tumor stem cell load | DC- |
| 1 | 1∶40 | 2.013 | 1.02 |
| 2 | 1∶20 | 1.966 | 0.905 |
| 3 | 1∶10 | 1.234 | 0.891 |
| 4 | 1∶5 | 0.612 | 0.714 |
| 5 | 1∶2.5 | 0.434 | 0.475 |
| 6 | 1∶1.25 | 0.378 | 0.312 |
| 7 | Control (contrast) | 0.396 | 0.436 |
From table 1 and Fig. 1, can draw, have the DC-CIK cell vaccine of tumor stem cell antigen load that target cell is had stronger lethality, and the effect target is higher than more, the corresponding enhancing of cell killing effect.
The result of this research shows that behind the DC sensitization CIK of load multi-drug resistant breast carcinoma stem cell, CIK has remarkable advantages to the killing activity of this breast carcinoma stem cell than matched group.After its mechanism possibly be DC load antigen, the various collaborative stimulation molecules of high expressed had strengthened identification and the specific killing of T cell to target cell.Simultaneously, the excretory various kinds of cell factor of ripe DC has promoted the activation of CIK and NK cell, has strengthened Th1 type immunne response in the body.DC and CIK co-culture of cells can strengthen the quantity and the killing activity of CIK cell greatly, improve the clinical efficacy of CIK cell therapy.Utilize the DC cell and the CIK cell of load tumor antigen to hatch altogether, can promote propagation and the immunologic function of cell and DC simultaneously, thereby the wide spectrum of performance CIK cell kills the double effect of tumor and the activated specificity antineoplastic of DC.
The present invention has also studied the lethal effect of the DC cell of load multi-drug resistant breast carcinoma stem cell to tumor cell, and (experimentation and aforementioned similar, matched group are selected the not antigenic DC-CIK of load tumor stem cell to adopt the classical tumor experiment of killing.Experimental result is seen table 2 and Fig. 2.
Table 2
| Sequence number | Imitate the target ratio | The DC of tumor stem cell load | DC- |
| 1 | 1∶40 | 1.312 | 1.02 |
| 2 | 1∶20 | 1.25 | 0.905 |
| 3 | 1∶10 | 1.001 | 0.891 |
| 4 | 1∶5 | 0.723 | 0.714 |
| 5 | 1∶2.5 | 0.451 | 0.475 |
| 6 | 1∶1.25 | 0.288 | 0.312 |
| 7 | Control (contrast) | 0.312 | 0.401 |
From table 2 and Fig. 2, can draw, have single DC cell vaccine of antigen load that target cell is also had stronger lethality, and the effect target is higher than more, the corresponding enhancing of cell killing effect.Certain and CIK cultivates altogether and can obtain better to kill the tumor effect.
Above-described embodiment describes preferred implementation of the present invention; Be not that scope of the present invention is limited; Design under the prerequisite of spirit not breaking away from the present invention; Various distortion and improvement that the common engineers and technicians in this area make technical scheme of the present invention all should fall in the definite protection domain of claims of the present invention.
Claims (10)
1. dendritic cell tumor vaccine, it is characterized in that: said dendritic cell tumor vaccine load has multi-drug resistant tumor stem cell antigen compsn.
2. dendritic cell tumor vaccine according to claim 1; It is characterized in that: said multi-drug resistant tumor stem cell antigen compsn is with obtaining tumor stem cell after the former generation separation of tumor tissues process tumor stem cell, the cultivation; Screen the tumor stem cell with multi-drug resistant through flow cytometer again, the full cellular antigens of process prepare and obtain at last.
3. dendritic cell tumor vaccine according to claim 2 is characterized in that: said tumor stem cell with multi-drug resistant is multidrug resistant gene and these two cells that surface marker is all positive of product P-glycoprotein thereof.
4. the method for preparing of the described dendritic cell tumor vaccine of claim 1 is characterized in that: may further comprise the steps:
(1) tumor tissues is carried out former generation separates and cultivation obtains tumor stem cell;
(2) adopt the flow cytometer screening to have the tumor stem cell of multi-drug resistant;
(3) tumor stem cell that step (2) is obtained prepares full cellular antigens;
(4) isolated dendritic cell and cultivation;
(5) the full cellular antigens that step (3) obtained and the BMDC of step (4) are cultivated altogether and obtain the dendritic cell tumor vaccine that load has multi-drug resistant tumor stem cell antigen compsn.
5. the method for preparing of the described dendritic cell tumor vaccine of claim 1 is characterized in that: may further comprise the steps:
(1) tumor tissues is carried out former generation separates and cultivation obtains tumor stem cell;
(2) adopt the flow cytometer screening to have the tumor stem cell of multi-drug resistant;
(3) tumor stem cell that step (2) is obtained prepares full cellular antigens;
(4) isolated dendritic cell and cytokine induced kill cell and cultivation;
(5) the full cellular antigens that step (3) obtained and the BMDC of step (4) and cytokine induced kill cell are cultivated altogether and are obtained the dendritic cell tumor vaccine that load has multi-drug resistant tumor stem cell antigen compsn.
6. according to claim 4 or 5 described method for preparinies, it is characterized in that: the tumor tissues in the said step (1) is a breast tumor tissues.
7. method for preparing according to claim 6; It is characterized in that: adopt flow cytometer that the breast tumor stem cell that cultivation obtains is carried out the detection of multidrug resistant gene and product P-glycoprotein thereof in the said step (2); Filter out this two cells that surface marker is all positive, be breast tumor stem cell with multi-drug resistant.
8. the dendritic cell tumor vaccine for preparing of claim 6 or 7 described method for preparinies.
9. the application of the described dendritic cell tumor vaccine of claim 1 in the medicine of preparation treatment tumor disease.
10. the application of the described dendritic cell tumor vaccine of claim 8 in the medicine of preparation treatment breast tumor disease.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105308177A (en) * | 2013-03-07 | 2016-02-03 | 新干细胞肿瘤学有限责任公司 | Individualized high purity hepatocellular carcinoma stem cells, methods and use of the same |
| WO2016036319A1 (en) * | 2014-09-04 | 2016-03-10 | Agency For Science, Technology And Research | A method to up-regulate cancer stem cell markers for the generation of antigen specific cytotoxic effector t cells |
| CN106811442A (en) * | 2015-12-01 | 2017-06-09 | 上海市中医老年医学研究所 | The preparation of DC-CIK cells and its application in treatment ovarian cancer is prepared |
| CN113440606A (en) * | 2021-06-02 | 2021-09-28 | 深圳市罗湖区人民医院 | Specific pluripotent stem cell tumor vaccine and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101538554A (en) * | 2009-04-30 | 2009-09-23 | 北京弘润天源生物技术有限公司 | Survivability-screened cancer stem cells, preparation method, application and kit as well as antigen composite thereof, preparation method, application and kit |
| CN101560496A (en) * | 2009-04-30 | 2009-10-21 | 北京弘润天源生物技术有限公司 | Dendritic cell carried by tumor stem cell antigen and subjected to tolerance screening, preparation method and application thereof, kit and vaccine comprising dendritic cell |
-
2011
- 2011-05-26 CN CN2011101377389A patent/CN102793912A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101538554A (en) * | 2009-04-30 | 2009-09-23 | 北京弘润天源生物技术有限公司 | Survivability-screened cancer stem cells, preparation method, application and kit as well as antigen composite thereof, preparation method, application and kit |
| CN101560496A (en) * | 2009-04-30 | 2009-10-21 | 北京弘润天源生物技术有限公司 | Dendritic cell carried by tumor stem cell antigen and subjected to tolerance screening, preparation method and application thereof, kit and vaccine comprising dendritic cell |
Non-Patent Citations (3)
| Title |
|---|
| CYNTHIA E, HERZOG J B, TREPEL L, ET AL.: "Various Methods of Analysis of mdr-1/P-Glycoprotein in Human Colon Cancer Cell Lines", 《J NATL CANCER INST》, vol. 84, no. 9, 6 May 1992 (1992-05-06) * |
| 侯金程等: "树突状细胞疫苗联合细胞因子诱导的杀伤细胞治疗鼻腔及鼻窦恶性黑色素瘤的近期疗效观察", 《临床医学》, vol. 30, no. 1, 31 January 2010 (2010-01-31) * |
| 曾飞跃 等: "MDR1在人脑肿瘤干细胞中的表达及意义", 《吉林大学学报（医学版）》, vol. 35, no. 5, 30 September 2009 (2009-09-30), pages 769 - 774 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105308177A (en) * | 2013-03-07 | 2016-02-03 | 新干细胞肿瘤学有限责任公司 | Individualized high purity hepatocellular carcinoma stem cells, methods and use of the same |
| EP2964754A4 (en) * | 2013-03-07 | 2016-08-17 | Neostem Oncology Llc | Individualized high purity hepatocellular carcinoma stem cells, methods and use of the same |
| WO2016036319A1 (en) * | 2014-09-04 | 2016-03-10 | Agency For Science, Technology And Research | A method to up-regulate cancer stem cell markers for the generation of antigen specific cytotoxic effector t cells |
| CN107148470A (en) * | 2014-09-04 | 2017-09-08 | 新加坡科技研究局 | Raise method of the cancer stem cell mark to produce antigen-specific cytotoxic effector T cell |
| CN106811442A (en) * | 2015-12-01 | 2017-06-09 | 上海市中医老年医学研究所 | The preparation of DC-CIK cells and its application in treatment ovarian cancer is prepared |
| CN106811442B (en) * | 2015-12-01 | 2020-12-29 | 上海市中医老年医学研究所 | Preparation of DC-CIK cells and application of DC-CIK cells in preparation of medicines for treating ovarian cancer |
| CN113440606A (en) * | 2021-06-02 | 2021-09-28 | 深圳市罗湖区人民医院 | Specific pluripotent stem cell tumor vaccine and preparation method and application thereof |
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