CN103815973A - Building method and applications of living tumor tissue bank - Google Patents
Building method and applications of living tumor tissue bank Download PDFInfo
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- CN103815973A CN103815973A CN201410083459.2A CN201410083459A CN103815973A CN 103815973 A CN103815973 A CN 103815973A CN 201410083459 A CN201410083459 A CN 201410083459A CN 103815973 A CN103815973 A CN 103815973A
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Abstract
The invention provides a building method and applications of a living tumor tissue bank. Tumor tissues of a cancerous patient subjected to an operation are transplanted to a part under the renal capsules of SCID (severe combined immunodeficient mice) and survive in the bodies of the mice for passage conservation, so that the living tumor tissue bank is obtained; the living tumor tissue bank means that based on single tumor tissue heterograft, numerous human body tumor tissues are conserved in animal bodies in a living tissue bank form for standby application; the living tumor tissue bank comprises transverse multiple tumors and various subtypes or various development phase of a certain longitudinal tumor. The living tumor tissue bank has multiple applications in the aspects of determining the personalized therapeutic schedule of the cancerous patient subjected to the operation, screening tumor specificities of anti-cancer drugs, and building animal models for human body cancer research.
Description
Technical field
The invention belongs to technology of cancer treatment field, relate in particular to a kind of method for building up and application thereof of vivo tumor tissue bank.
Background technology
Cancer is on the rise to the mankind's harm, in average every four dead patients, has at least one to die from cancer.The current passive passive state for treatment of cancer is mainly owing to two aspects: the one, and the complexity of cancer itself and contingency (comprising the Drug resistance to medicine) highly; The 2nd, unicity (without specificity and directionality) and the immutableness for the treatment of measure.A desirable treatment measure should act on tumor tissues in specific manner, reach with multipair assorted, with variable to changeable, cancer and the assembly of this contradiction for the treatment of are organically combined on human body individual character, tumor characteristic, cell and even molecular characterization are at all levels.Cancer with treatment measure dynamic monitoring be achieve the above object indispensable or precondition.
Cancer worker early has recognized that the importance in tumor tissues storehouse, has worldwide set up (or setting up) various scales, polytype body tumor tissue storehouse.Tumor tissues storehouse is most important to the research of cancer as valuable resource.But the information obtaining from this traditional tissue bank is to have existed tumor tissues and is static, can not obtain the multidate information organically combining with therapeutic scheme.
Summary of the invention
The object of the present invention is to provide a kind of method for building up and application thereof of vivo tumor tissue bank, be intended to solve traditional tissue bank and can not obtain the multidate information organically combining with therapeutic scheme, effective treatment approach for exploring with practical significance provides strong experiment porch.
The present invention is achieved in that a kind of method for building up of vivo tumor tissue bank, comprises the following steps:
(1) by the object tumor tissues pretreatment cutting;
(2) described tumor tissues is migrated under mouse kidney tunicle;
(3) after tumor growth arrives pre-sizing, put to death mice, under aseptic condition, expose tumor, cut tumor tissues after obtaining image data, the Partial tumors tissue that mice is obtained is with it divided into two groups, and wherein one group is carried out stored frozen;
(4) another group tumor tissues repeating step (1) and (2) is carried out to biological tissue and go down to posterity, obtain vivo tumor tissue bank; Or biological tissue is carried out in repeating step (1) and (2) after the tumor tissues recovery of described stored frozen and go down to posterity, obtain vivo tumor tissue bank.
Preferably, in step (1), described tumor tissues pretreatment comprises: the body tumor tissue of excision is placed in aseptic, cryopreservation solution immediately, removes identifiable slough; Vigorous tumor growth position is cut into the piece of tissue of 1 × 1 × 3mm.
Preferably, described human tumor is any human body solid malignant, comprises pulmonary carcinoma, hepatocarcinoma, gastric cancer, esophageal carcinoma, nasopharyngeal carcinoma, cancer of pancreas, colon cancer, bladder cancer, renal carcinoma, skin carcinoma, thyroid carcinoma, bladder cancer, breast carcinoma, cervical cancer, ovarian cancer, carcinoma of prostate, glioblastoma, osteosarcoma.
Preferably, in step (2), described mice is the Mus SCID mice in 4～5 weeks ages.
Preferably, in step (2), described transplanting comprises: mice is exposed to left kidney under routine disinfection, anesthesia, provoke kidney tunicle in kidney ridge upper end with injection needle, cut an osculum, then peel off kidney tunicle to kidney ridge lower end with little Glass rod, tumor mass is put in to each one of low side and upper end with pincet; Finally kidney reset, respectively suture muscles and skin, routine recovered and nurse.
Preferably, in step (3), described stored frozen comprises: tumor tissues is placed in the preservation liquid that contains 1%DMSO, the speed of spending with reduction by 1 per minute is down to-80 ℃, transfers in liquid nitrogen and preserve after 3 hours.
Preferably, in step (4), described tumor tissues recovery comprises: the tumor tissues of stored frozen is placed in to 37 ℃ of water-baths and recovers.
The present invention further provides the application aspect personalized therapy program after definite cancer patient performs the operation of above-mentioned vivo tumor tissue bank.
The present invention further provides the application of above-mentioned vivo tumor tissue bank aspect the tumour-specific screening of cancer therapy drug.
The present invention further provides the application of above-mentioned vivo tumor tissue bank aspect the animal model of structure human body cancer research.
The present invention overcomes the deficiencies in the prior art, a kind of method for building up and application thereof of vivo tumor tissue bank are provided, by postoperative human body cancer patient tumor tissues being transplanted under the kidney tunicle of SCID mice (Severe combined immunodeficient mice), in Mice Body, survive and go down to posterity preservation, obtain vivo tumor tissue bank, vivo tumor tissue bank is on single tumor tissues xenotransplantation basis, by numerous body tumor tissues with the form of biological tissue storehouse (Living tissue bank) preserve in animal body, for subsequent use.This vivo tumor tissue bank comprises horizontal kinds of tumors, also comprises various hypotypes or the various developmental stage of certain longitudinal tumor.This vivo tumor tissue bank serves many purposes, be included in determine cancer patient perform the operation after the application of personalized therapy program aspect, the application aspect the tumour-specific screening of cancer therapy drug, and in the application building aspect the animal model of human body cancer research.
Accompanying drawing explanation
Fig. 1 is the medium and small Ren Mus tunicle of comparative example of the present invention and Subcutaneous tumor tissue growth situation pictorial diagram;
Fig. 2 be in purposes embodiment 1 of the present invention vivo tumor tissue bank for determining the technology path schematic diagram of Personalized Ways of cancer patient's post-operative treatment;
Fig. 3 is the technology path schematic diagram that in purposes embodiment 2 of the present invention, vivo tumor tissue bank screens for the tumour-specific of cancer therapy drug;
Fig. 4 be in purposes embodiment 3 of the present invention vivo tumor tissue bank for building the technology path schematic diagram of animal model of human body cancer research.
The specific embodiment
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.
1, the foundation of vivo tumor tissue bank and correlation technique
Animal is selected: SCID mice (Severe combined immunodeficient mice), male or female, 4～5 weeks ages of Mus.
2, tumor is selected: any human body solid malignant, comprises pulmonary carcinoma, hepatocarcinoma, gastric cancer, esophageal carcinoma, nasopharyngeal carcinoma, cancer of pancreas, colon cancer, bladder cancer, renal carcinoma, skin carcinoma, thyroid carcinoma, bladder cancer, breast carcinoma, cervical cancer, ovarian cancer, carcinoma of prostate, glioblastoma, osteosarcoma etc.
3, Data Base Pile is set up: the general state of an illness data that comprises (1) patient; (2) tumor special examined data; (3) tumor tissues storehouse related data.
4, the foundation of vivo tumor tissue bank:
(1) organized processing: the tumor tissues of excision is placed in aseptic, cryopreservation solution immediately, prunes, remove identifiable slough in biological cleaning platform.Vigorous tumor growth position is cut into the piece of tissue of 1 × 1 × 3mm, gets 4～6 block organizations for transplanting, 3～4 block organization's stored frozen depending on the size of tissue.This process, in ice surface, sterile working, should complete in 30～60 minutes.
(2) tissue transplantation: mice is exposed to left kidney under routine disinfection, anesthesia, provoke kidney tunicle, cut an osculum with injection needle in kidney ridge upper end, then peel off kidney tunicle to kidney ridge lower end with little Glass rod, tumor mass is put in to each one of low side and upper end with pincet.Finally kidney reset, respectively suture muscles and skin, routine recovered and nurse.To transplant first and be defined as for 0 generation.
(3) tissue goes down to posterity: arrive external visible or when tangible until tumor growth, routine is put to death animal, under aseptic condition, is exposed tumor, obtain tumor resection after image data, then re-start tissue transplantation according to above-mentioned steps (1) and (2), obtain vivo tumor tissue bank.The number of times that tissue goes down to posterity, depending on follow-up needs, is preserved 1 generation tissue (6～12) as tissue bank.
(4) tissue is preserved: piece of tissue is placed in the preservation liquid that contains 1%DMSO, the speed of spending with reduction by 1 per minute is down to-80 ℃, transfers in liquid nitrogen and preserve after 3 hours.
(5) tissue recovery: by preserve 1 generation tissue be placed in 37 ℃ of water-baths and recover, after conventional cleaning, transplant (1) and (2) according to the method described above, obtains vivo tumor tissue bank.
Comparative example
Each step of this comparative example is all identical with the step in embodiment 1, and difference is, the present embodiment is that tumor mass is transplanted to the subcutaneous of mice.Tumor size in comparing embodiment 1 and comparative example, as shown in Figure 1, wherein, figure a is initial glioma cells in tissue piece, figure b-1 cultivates after 50 days tumor tissues under Mouse Kidney tunicle in embodiment 1, figure b-2 cultivates mice Subcutaneous tumor tissue after 40 days in comparative example.As seen from Figure 1, onesize glioma cells in tissue is transplanted to respectively under the subcutaneous of mice and kidney tunicle, within 40 days, is found afterwards that the tumor of transplanting under kidney tunicle is obviously greater than the tumor of subcutaneous transplantation.
The vivo tumor tissue bank that embodiment 1 sets up is for determining the Personalized Ways of cancer patient's post-operative treatment, as shown in Figure 2, as shown in Figure 2, postoperative certain cancer patient tumor tissues is transplanted under SCID Mouse Kidney tunicle, press the different schemes treatments such as A in Fig. 1, B, C, observe different therapeutic schemes tumor growth is suppressed, killed and wounded and prognosis texts, thereby determine that C is best personalized therapy program.
Purposes embodiment 2
The vivo tumor tissue bank that embodiment 1 sets up is for the tumour-specific screening of cancer therapy drug, as shown in Figure 3, as shown in Figure 3, the tumor tissues of variety classes cancer is transplanted under same SCID Mouse Kidney tunicle, can be guaranteed that all experimental subjecies monitor under on all four condition.As shown in Figure 3, accept after monitored tumor pharmacother, nasopharyngeal carcinoma (◆) and cerebral glioma (
) continued growth becomes large, pulmonary carcinoma (
) there is no significant change, and hepatocarcinoma (●) obviously diminishes or disappears, and illustrates that this tumour medicine is suitable for the treatment of hepatocarcinoma most.
Purposes embodiment 3
The vivo tumor tissue bank that embodiment 1 sets up is for building the animal model of human body cancer research, as shown in Figure 4, as shown in Figure 4, for better simulation cancer is in people's tumor growth, transfer invasion and attack situation, the body tumor tissue of separate sources is carried out to original position plantation, in figure by hepatocarcinoma (●) pulmonary carcinoma in vivo tumor tissue bank (
) and cerebral glioma (
) tissue is transplanted to respectively in liver, lungs and brain, sets up corresponding cancer in situ animal model.Cancer in situ animal model can be used for the research of cancer generation, development and cancer metastasis mechanism, also can be for the experimentation of antitumor drug and special treatment approach research and development aspect.
Compare and the shortcoming and defect of prior art, the present invention has following beneficial effect: vivo tumor tissue bank of the present invention is easy to build, and the tumor growth of going down to posterity retains all right, remarkably productive; Vivo tumor tissue bank of the present invention serves many purposes, be included in determine cancer patient perform the operation after the application of personalized therapy program aspect, application aspect the tumour-specific screening of cancer therapy drug, and in the application building aspect the animal model of human body cancer research.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any modifications of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.
Claims (10)
1. a method for building up for vivo tumor tissue bank, is characterized in that comprising the following steps:
(1) by the object tumor tissues pretreatment cutting;
(2) described tumor tissues is migrated under mouse kidney tunicle;
(3) after tumor growth arrives pre-sizing, put to death mice, under aseptic condition, expose tumor, cut tumor tissues after obtaining image data, the Partial tumors tissue that mice is obtained is with it divided into two groups, and wherein one group is carried out stored frozen;
(4) another group tumor tissues repeating step (1) and (2) is carried out to biological tissue and go down to posterity, obtain vivo tumor tissue bank; Or biological tissue is carried out in repeating step (1) and (2) after the tumor tissues recovery of described stored frozen and go down to posterity, obtain vivo tumor tissue bank.
2. the method for building up of vivo tumor tissue bank as claimed in claim 1, it is characterized in that, in step (1), described tumor tissues pretreatment comprises: the body tumor tissue of excision is placed in aseptic, cryopreservation solution immediately, removes identifiable slough; Vigorous tumor growth position is cut into the piece of tissue of 1 × 1 × 3mm.
3. the method for building up of vivo tumor tissue bank as claimed in claim 2, it is characterized in that, described human tumor is any human body solid malignant, comprises pulmonary carcinoma, hepatocarcinoma, gastric cancer, esophageal carcinoma, nasopharyngeal carcinoma, cancer of pancreas, colon cancer, bladder cancer, renal carcinoma, skin carcinoma, thyroid carcinoma, bladder cancer, breast carcinoma, cervical cancer, ovarian cancer, carcinoma of prostate, glioblastoma, osteosarcoma.
4. the method for building up of vivo tumor tissue bank as claimed in claim 3, is characterized in that, in step (2), described mice is the Mus SCID mice in 4～5 weeks ages.
5. the method for building up of vivo tumor tissue bank as claimed in claim 4, it is characterized in that, in step (2), described transplanting comprises: mice is exposed to left kidney under routine disinfection, anesthesia, provoke kidney tunicle in kidney ridge upper end with injection needle, cut an osculum, then peel off kidney tunicle to kidney ridge lower end with little Glass rod, tumor mass is put in to each one of low side and upper end with pincet; Finally kidney reset, respectively suture muscles and skin, routine recovered and nurse.
6. the method for building up of vivo tumor tissue bank as claimed in claim 5, it is characterized in that, in step (3), described stored frozen comprises: tumor tissues is placed in the preservation liquid that contains 1%DMSO, the speed of spending with reduction by 1 per minute is down to-80 ℃, transfers in liquid nitrogen and preserve after 3 hours.
7. the method for building up of vivo tumor tissue bank as claimed in claim 6, is characterized in that, in step (4), described tumor tissues recovery comprises: the tumor tissues of stored frozen is placed in to 37 ℃ of water-baths and recovers.
8. the application aspect personalized therapy program after definite cancer patient performs the operation of the vivo tumor tissue bank described in claim 1 to 7 any one.
9. the application of the vivo tumor tissue bank described in claim 1 to 7 any one aspect the tumour-specific screening of cancer therapy drug.
10. the application of the vivo tumor tissue bank described in claim 1 to 7 any one aspect the animal model of structure human body cancer research.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105696087A (en) * | 2015-12-31 | 2016-06-22 | 四川大学华西医院 | Lung cancer PDX standardized model library |
| CN105684989A (en) * | 2015-12-31 | 2016-06-22 | 四川大学华西医院 | Liver cancer PDX standardized model library |
| CN110402895A (en) * | 2019-09-09 | 2019-11-05 | 中南大学湘雅二医院 | A kind of construction method of osteosarcoma xenograft mouse model |
| CN110476892A (en) * | 2019-09-09 | 2019-11-22 | 中南大学湘雅二医院 | A kind of construction method of osteosarcoma xenograft mouse model and application |
| CN110495422A (en) * | 2019-09-23 | 2019-11-26 | 中南大学湘雅二医院 | A kind of construction method of malignant schwannoma xenograft mouse model and its application |
| CN110495424A (en) * | 2019-09-09 | 2019-11-26 | 中南大学湘雅二医院 | A kind of construction method of synovial sarcoma xenograft mouse model |
| CN110547250A (en) * | 2019-09-23 | 2019-12-10 | 中南大学湘雅二医院 | Construction method and application of synovial sarcoma xenograft mouse model with healthy immunity |
| CN110623770A (en) * | 2019-09-23 | 2019-12-31 | 中南大学湘雅二医院 | Method for constructing malignant schwannoma xenograft mouse model |
| CN110637783A (en) * | 2019-09-23 | 2020-01-03 | 中南大学湘雅二医院 | Construction method of synovial sarcoma xenograft mouse model with healthy immunity |
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2014
- 2014-03-07 CN CN201410083459.2A patent/CN103815973A/en active Pending
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105696087A (en) * | 2015-12-31 | 2016-06-22 | 四川大学华西医院 | Lung cancer PDX standardized model library |
| CN105684989A (en) * | 2015-12-31 | 2016-06-22 | 四川大学华西医院 | Liver cancer PDX standardized model library |
| CN110402895A (en) * | 2019-09-09 | 2019-11-05 | 中南大学湘雅二医院 | A kind of construction method of osteosarcoma xenograft mouse model |
| CN110476892A (en) * | 2019-09-09 | 2019-11-22 | 中南大学湘雅二医院 | A kind of construction method of osteosarcoma xenograft mouse model and application |
| CN110495424A (en) * | 2019-09-09 | 2019-11-26 | 中南大学湘雅二医院 | A kind of construction method of synovial sarcoma xenograft mouse model |
| CN110495422A (en) * | 2019-09-23 | 2019-11-26 | 中南大学湘雅二医院 | A kind of construction method of malignant schwannoma xenograft mouse model and its application |
| CN110547250A (en) * | 2019-09-23 | 2019-12-10 | 中南大学湘雅二医院 | Construction method and application of synovial sarcoma xenograft mouse model with healthy immunity |
| CN110623770A (en) * | 2019-09-23 | 2019-12-31 | 中南大学湘雅二医院 | Method for constructing malignant schwannoma xenograft mouse model |
| CN110637783A (en) * | 2019-09-23 | 2020-01-03 | 中南大学湘雅二医院 | Construction method of synovial sarcoma xenograft mouse model with healthy immunity |
| CN110547250B (en) * | 2019-09-23 | 2021-12-28 | 中南大学湘雅二医院 | Construction method and application of synovial sarcoma xenograft mouse model with healthy immunity |
| CN110637783B (en) * | 2019-09-23 | 2021-12-28 | 中南大学湘雅二医院 | Construction method of synovial sarcoma xenograft mouse model with healthy immunity |
| CN110623770B (en) * | 2019-09-23 | 2022-12-09 | 中南大学湘雅二医院 | Construction method of malignant schwannoma xenograft mouse model |
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Application publication date: 20140528 |