CN108048401A - People's biliary tract cancerous cell line and application - Google Patents
People's biliary tract cancerous cell line and application Download PDFInfo
- Publication number
- CN108048401A CN108048401A CN201810004140.4A CN201810004140A CN108048401A CN 108048401 A CN108048401 A CN 108048401A CN 201810004140 A CN201810004140 A CN 201810004140A CN 108048401 A CN108048401 A CN 108048401A
- Authority
- CN
- China
- Prior art keywords
- people
- cell line
- zju
- biliary tract
- cancerous cell
- 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
- 210000003445 biliary tract Anatomy 0.000 title claims abstract description 64
- 206010004593 Bile duct cancer Diseases 0.000 claims abstract description 17
- 210000000232 gallbladder Anatomy 0.000 claims abstract description 11
- 208000007666 Klatskin Tumor Diseases 0.000 claims abstract description 10
- 208000018060 hilar cholangiocarcinoma Diseases 0.000 claims abstract description 10
- 210000004027 cell Anatomy 0.000 claims description 143
- 210000000013 bile duct Anatomy 0.000 claims description 17
- 206010028980 Neoplasm Diseases 0.000 claims description 16
- 208000026900 bile duct neoplasm Diseases 0.000 claims description 15
- 210000000349 chromosome Anatomy 0.000 claims description 12
- 241000204031 Mycoplasma Species 0.000 claims description 9
- 238000001514 detection method Methods 0.000 claims description 9
- 201000011510 cancer Diseases 0.000 claims description 8
- 230000007812 deficiency Effects 0.000 claims description 8
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 claims description 8
- 229960005277 gemcitabine Drugs 0.000 claims description 8
- 230000000973 chemotherapeutic effect Effects 0.000 claims description 7
- 238000011580 nude mouse model Methods 0.000 claims description 6
- 238000011109 contamination Methods 0.000 claims description 5
- 230000007246 mechanism Effects 0.000 claims description 5
- 241000124008 Mammalia Species 0.000 claims description 4
- 201000010175 gallbladder cancer Diseases 0.000 claims description 4
- 210000004907 gland Anatomy 0.000 claims description 4
- 238000004321 preservation Methods 0.000 claims description 4
- 108091092878 Microsatellite Proteins 0.000 claims description 3
- 230000002159 abnormal effect Effects 0.000 claims description 3
- 239000003153 chemical reaction reagent Substances 0.000 claims description 3
- 201000007487 gallbladder carcinoma Diseases 0.000 claims description 3
- 230000005740 tumor formation Effects 0.000 claims description 3
- 210000003237 epithelioid cell Anatomy 0.000 claims description 2
- 210000000981 epithelium Anatomy 0.000 claims description 2
- 230000002401 inhibitory effect Effects 0.000 claims description 2
- 230000003211 malignant effect Effects 0.000 claims description 2
- 238000003745 diagnosis Methods 0.000 claims 1
- 238000011160 research Methods 0.000 abstract description 6
- 230000008506 pathogenesis Effects 0.000 abstract description 2
- 238000006243 chemical reaction Methods 0.000 description 16
- 239000000243 solution Substances 0.000 description 14
- 108020004414 DNA Proteins 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 11
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- 239000013641 positive control Substances 0.000 description 7
- 239000006285 cell suspension Substances 0.000 description 6
- 210000002950 fibroblast Anatomy 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 108010019160 Pancreatin Proteins 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 238000001962 electrophoresis Methods 0.000 description 5
- 229940088598 enzyme Drugs 0.000 description 5
- 229940055695 pancreatin Drugs 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 102000001400 Tryptase Human genes 0.000 description 4
- 108060005989 Tryptase Proteins 0.000 description 4
- 238000001976 enzyme digestion Methods 0.000 description 4
- 235000019441 ethanol Nutrition 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 238000001356 surgical procedure Methods 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 102000029816 Collagenase Human genes 0.000 description 3
- 108060005980 Collagenase Proteins 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 241000699660 Mus musculus Species 0.000 description 3
- 239000012980 RPMI-1640 medium Substances 0.000 description 3
- 108010087230 Sincalide Proteins 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 201000009036 biliary tract cancer Diseases 0.000 description 3
- 208000020790 biliary tract neoplasm Diseases 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 238000010609 cell counting kit-8 assay Methods 0.000 description 3
- 229960002424 collagenase Drugs 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- IAKHMKGGTNLKSZ-INIZCTEOSA-N (S)-colchicine Chemical compound C1([C@@H](NC(C)=O)CC2)=CC(=O)C(OC)=CC=C1C1=C2C=C(OC)C(OC)=C1OC IAKHMKGGTNLKSZ-INIZCTEOSA-N 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 2
- 208000005623 Carcinogenesis Diseases 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 102100028999 High mobility group protein HMGI-C Human genes 0.000 description 2
- 101000986379 Homo sapiens High mobility group protein HMGI-C Proteins 0.000 description 2
- 102000002151 Microfilament Proteins Human genes 0.000 description 2
- 108010040897 Microfilament Proteins Proteins 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- 230000036952 cancer formation Effects 0.000 description 2
- 231100000504 carcinogenesis Toxicity 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 208000006990 cholangiocarcinoma Diseases 0.000 description 2
- 230000002559 cytogenic effect Effects 0.000 description 2
- 210000001047 desmosome Anatomy 0.000 description 2
- 230000001079 digestive effect Effects 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 230000009977 dual effect Effects 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 238000009415 formwork Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 206010017758 gastric cancer Diseases 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 210000003632 microfilament Anatomy 0.000 description 2
- 210000003470 mitochondria Anatomy 0.000 description 2
- 210000003463 organelle Anatomy 0.000 description 2
- 210000000496 pancreas Anatomy 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 210000003705 ribosome Anatomy 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 201000011549 stomach cancer Diseases 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 230000017105 transposition Effects 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 208000022072 Gallbladder Neoplasms Diseases 0.000 description 1
- -1 GelRed nucleic acid Chemical class 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 241001062009 Indigofera Species 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 206010028470 Mycoplasma infections Diseases 0.000 description 1
- 241000204003 Mycoplasmatales Species 0.000 description 1
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 1
- 208000026487 Triploidy Diseases 0.000 description 1
- JDZJVWAHZYIHFA-UHFFFAOYSA-N [Br].C1(=CC=CC=C1)O Chemical compound [Br].C1(=CC=CC=C1)O JDZJVWAHZYIHFA-UHFFFAOYSA-N 0.000 description 1
- SFCVEUVVOJFYSX-UHFFFAOYSA-J [Pb+2].C(CC(O)(C(=O)[O-])CC(=O)[O-])(=O)[O-].C(C)(=O)[O-].[U+6] Chemical compound [Pb+2].C(CC(O)(C(=O)[O-])CC(=O)[O-])(=O)[O-].C(C)(=O)[O-].[U+6] SFCVEUVVOJFYSX-UHFFFAOYSA-J 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 239000000980 acid dye Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012267 brine Substances 0.000 description 1
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 238000009104 chemotherapy regimen Methods 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 229960001338 colchicine Drugs 0.000 description 1
- 238000012864 cross contamination Methods 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 239000012531 culture fluid Substances 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000012137 double-staining Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 210000001501 megacaryocyte Anatomy 0.000 description 1
- 230000031864 metaphase Effects 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 210000000110 microvilli Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 210000000633 nuclear envelope Anatomy 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000002559 palpation Methods 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000003934 vacuole Anatomy 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
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
- C12N5/0693—Tumour cells; Cancer cells
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/12—Animals modified by administration of exogenous cells
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/0331—Animal model for proliferative diseases
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Wood Science & Technology (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Environmental Sciences (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Cell Biology (AREA)
- General Health & Medical Sciences (AREA)
- Oncology (AREA)
- Animal Behavior & Ethology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Microbiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to technical field of microbe cell line, and in particular to people's biliary tract cancerous cell line and application.People's biliary tract cancerous cell line, including：Gallbladder cell-Shandong ZJU 0430, people's hilar cholangiocarcinoma ZJU 0826 and people's hilar cholangiocarcinoma ZJU 1125 are respectively designated as, and is preserved in China typical culture collection center, deposit number is respectively：CCTCC NO:C2017174、CCTCC NO:C2017175 and CCTCC NO:People's biliary tract cancerous cell line of C2017176.People's biliary tract cancerous cell line ZJU 0430, ZJU 0826 and the ZJU 1125 of the present invention is established from Chinese source, and it is that the time is shorter to build, biological heredity character is stablized, using the biliary tract cancerous cell line as research model, have very great help for the primary biliary cancer pathogenesis for understanding Chinese population.
Description
Technical field
The invention belongs to technical field of microbe cell line, and in particular to people's biliary tract cancerous cell line and application.
Background technology
Although being improved at present in diagnose and treat technical aspect, cancer of bile ducts patient's survival region is poor.Previously
Research report surgery excision be to treat the sick unique method, there is no effective chemotherapy regimen for not performing the operation or postoperative recurrence
Patient.The essence for exploring the oncobiology is particularly important for the prognostic for improving cancer of bile ducts patient.At the same time, exist at present
The bad present situation of cancer of bile ducts therapeutic effect, one of main cause are a lack of perfect culture system in vitro.However current body
The tumour cell of outer culture ties up to many fields such as tumorigenesis, invasion and attack, metastasis and early diagnosis, treatment evaluation
Research in play irreplaceable role.
The content of the invention
First purpose of the present invention is, the biliary tract cancerous cell line in Chinese population source is lacked for the current country, especially
It is the people's biliary tract cancerous cell line that can be applied to immune nude mice into knurl, and provide the type from Chinese population cancer of bile ducts
Cell line.
For this purpose, the above-mentioned purpose of the present invention is achieved through the following technical solutions：
People's biliary tract cancerous cell line, including：It is thin to be respectively designated as Gallbladder cell-Shandong ZJU-0430, people's hilar cholangiocarcinoma
Born of the same parents system ZJU-0826 and people hilar bile duct cancerous cell line ZJU-1125, and it is preserved in China typical culture collection center
(CCTCC), deposit number is respectively：CCTCC NO:C2017174、CCTCC NO:C2017175 and CCTCC NO:C2017176
People's biliary tract cancerous cell line.
While using above-mentioned technical proposal, the present invention can also be used or combined using technology further below
Scheme：
Preferably, people's biliary tract cancerous cell line further includes：It is respectively designated as Gallbladder cell-Shandong ZJU-0430, people liver
Door portion cholangiocarcinoma cell system ZJU-0826 and people hilar bile duct cancerous cell line ZJU-1125, and it is preserved in Chinese Typical Representative culture
Collection, deposit number are respectively：CCTCC NO:C2017174、CCTCC NO:C2017175 and CCTCC NO:
The daughter cell system of people's biliary tract cancerous cell line of C2017176.
Preferably, it is typical polygon epithelioid cell when people's biliary tract cancerous cell line is cultivated in vitro, under light microscopic,
It is typical Malignant Epithelium feature under Electronic Speculum.
Preferably, people's biliary tract cancerous cell line is gland cancer in cancer of bile ducts clinical diagnosis, and transplanted tumor in nude mice pathology is shown
For gland cancer；Karyotyping the results show its occur in chromosome number and structure it is abnormal.
Preferably, people's biliary tract cancerous cell line is in chemotherapeutic Gemcitabine (gemcitabine) Inhibition test, only people
Hilar bile duct cancerous cell line ZJU-0826 has partial inhibitory effect in higher concentrations to Gemcitabine.
Preferably, for people's biliary tract cancerous cell line when mycoplasma test reagent box detects, PCR results are feminine gender, not
It is subject to mycoplasma contamination；Karyotyping result be Gallbladder cell-Shandong ZJU-0430 chromosome numbers be 70-74, people's Hilar
Cholangiocarcinoma cell system ZJU-0826 chromosome numbers are 77-80 and people's hilar bile duct cancerous cell line ZJU-1125 chromosome numbers
For 58-69.
Second object of the present invention is, for the deficiencies in the prior art, provides people's biliary tract cancerous cell line
Using.
For this purpose, the above-mentioned purpose of the present invention is achieved through the following technical solutions：
According to application of the people's biliary tract cancerous cell line noted earlier in the cell model of cancer of bile ducts study of incident mechanism.
A further object of the invention is, for the deficiencies in the prior art, provides a kind of people's biliary tract cancer cell
The purposes of system.
For this purpose, the above-mentioned purpose of the present invention is achieved through the following technical solutions：
A kind of purposes of people's biliary tract cancerous cell line is used according to people's biliary tract cell line ZJU-0430 and ZJU-1125 noted earlier
In the model for preparing the generation cancer of bile ducts in immune deficiency mammal, tumor formation rate 100%.
While using above-mentioned technical proposal, the present invention can also be used or combined using technology further below
Scheme：
Preferably, the immune deficiency mammal is immune deficiency nude mouse.
Gallbladder cell-Shandong ZJU-0430 people sources are in an example stomach cancer and gall-bladder in people's biliary tract cancerous cell line of the present invention
Proteins in Carcinoma of Gallbladder sample, the people's hilar bile duct cancerous cell line ZJU-0826 of 69 years old male patient's surgery excision of cancer are derived from from one
The hilar cholangiocarcinoma sample and people's hilar bile duct cancerous cell line ZJU-1125 of 66 years old female patient surgery excision of example
Come from the Hepatic hilar tumor tissue specimen from 57 years old female patient surgery excision of an example hilar cholangiocarcinoma.Through original cuiture and build
After being tied to form work(, ZJU-0430, ZJU-0826 and ZJU-1125 are respectively designated as.
People's biliary tract cancerous cell line ZJU-0430, ZJU-0826 and ZJU-1125 cell in-vitro growth vigor of the present invention is high,
Cell mostly in flat irregular polygon, is grown, Chang Kejian vacuoles in endochylema in adherent monolayers；Cell after cryopreservation resuscitation still
With good vigor；Above-mentioned cell line has passed on or so 110 generations at present, can immortalize.
People's biliary tract cancerous cell line ZJU-0430, ZJU-0826 and ZJU-1125 cell line of the present invention and its daughter cell system
Cell doubling time respectively may be about for 24 hours, for 24 hours, 48h.Results of karyotype analysis shows that：Chromosome number and structure occur different
Often, chromosome number is distributed between 58-80 mostly, and textural anomaly includes chromosome increase, missing and transposition etc.；It is shown under light microscopic
Show that nuclear division is mutually apparent active, karyon is big, caryoplasm ratio increase, it is seen that megacaryocyte；Typical epithelial character under Electronic Speculum, can
See that desmosome, microfilament, abundant organelle (mitochondria, ribosomes and rough surfaced endoplasmic reticulum (RER)) and nucleocytoplasmic ratio are big.
People's biliary tract cancerous cell line of the present invention uses detection method most authoritative at present as ATCC (American Type Culture collection
Warehousing, American type culture collection) recommend STR (short tandem repeat, Short Tandem
Repeat) detection method, testing result determine that three people's biliary tract cancerous cell lines are people sources；With two whole world of ATCC and DSMZ most
Cytogenetics information in famous culture collection mechanism is compared, and discovery is not affected by other cell contaminations.
People's biliary tract cancerous cell line of the present invention is to chemotherapeutic Gemcitabine sensitivity Detections, finder's hilar bile duct
Cancerous cell line ZJU-0826 occurs dead with the raising of drug concentration, cellular portions；And other two kinds of cell line Human gallbladder carcinomas
Cell line ZJU-0430 and people's hilar bile duct cancerous cell line ZJU-1125 vigor do not influence.
People's biliary tract cancerous cell line of the present invention finds that the cell line established is in addition to ZJU-0826 naked into knurl experimental result
Good external Tumor formation is respectively provided in mouse body.By by a certain number of above-mentioned people's biliary tract cancerous cell line ZJU-0430 and ZJU-
1125 cell inoculations establish the animal model of cancer of bile ducts in the subcutaneous of nude mouse.
The present invention provides the daughter cell of the biliary tract cancerous cell line, the daughter cell is basic or all retains
The characteristic of parental cell.
Invention further provides people's biliary tract cancerous cell line in the cell model as cancer of bile ducts genesis mechanism research
In application.Since people's biliary tract cancerous cell line ZJU-0430, ZJU-0826 and ZJU-1125 of the present invention are from Chinese source
It establishes, and it is that the time is shorter to build, biological heredity character is stablized, using the biliary tract cancerous cell line as research model, for understanding
The pathogenesis of the primary biliary cancer of Chinese population has very great help.
In another aspect, the present invention provides the biliary tract cancer cells that different patient sources are established in a kind of cancer of bile ducts sample tissue
The preparation method of system, comprises the following steps：
(1) method of collection of specimens and preservation：Determine position of the lump in the biliary tract tissue of excision, tumor tissues are derived from
The surface part of knurl body active proliferation.The acquisition of sample carries out under pathologists guidance, prevents from examining pathological replacement
It is disconnected to have an impact；It when sample wouldn't do subsequent operation, can save it in 4 DEG C of RPMI 1640, can generally place left for 24 hours
It is right.
(2) original cuiture：Through PBS washings three times, machinery shreds tissue, collagenase digesting, and centrifugation and washing are inoculated with and pass
It is commissioned to train and supports and observe culture.
(3) purifying cells：Multiple pancreas enzyme -EDTA differential digestion is taken to obtain tumour cell, specially treats that cell closely converges
When, culture medium is abandoned, PBS is washed three times, adds in pancreas enzyme -EDTA digestive juice, Microscopic observation epithelial cell and fibroblast exist
The postdigestive variation of pancreatin, fibroblast is more sensitive to pancreatin, when spindle shape variation ellipse shape is even circular, adds in containing serum
New culture terminate digestion.
(4) if Microscopic observation also has the feasible multiple digestion removal fibroblast of fibroblast.
Description of the drawings
The optical microscope of Fig. 1 behaviour biliary tract cancerous cell line ZJU-0430, ZJU-0826 and ZJU-1125 cells；
The transmission electron microscope picture of Fig. 2 behaviour biliary tract cancerous cell line ZJU-0430, ZJU-0826 and ZJU-1125 cells；
The cell growth curve figure of Fig. 3 behaviour biliary tract cancerous cell line ZJU-0430, ZJU-0826 and ZJU-1125 cells；
Fig. 4 behaviour biliary tract cancerous cell line ZJU-0430, ZJU-0826 and ZJU-1125 detection of mycoplasma；
Fig. 5 behaviour biliary tract cancerous cell line ZJU-0430, ZJU-0826 and ZJU-1125 cells are to chemotherapeutic Gemcitabine
Sensitivity experiments；
The representative single celled karyotyping of Fig. 6 behaviour biliary tract cancerous cell lines ZJU-0430, ZJU-0826 and ZJU-1125；
Fig. 7 behaviour biliary tract cancerous cell line ZJU-0430, ZJU-0826 and ZJU-1125STR qualification figures；
The Tumorigenesis of Fig. 8 behaviour biliary tract cancerous cell line ZJU-0430 and ZJU-1125 cells.
Specific embodiment
The present invention is described in further detail with specific embodiment referring to the drawings.
Embodiment 1
From 69 years old male patient of an example stomach cancer and gallbladder cancer, 66 years old property patient of an example hilar cholangiocarcinoma and an example Hilar
Cholangiocarcinoma samples for 57 years old in female patient in situ tumor tissue, the tissues such as necrosis and fat is rejected, with containing dual anti-physiological saline
It washes 3 times.Tissue is inserted in sterile petri dish, a little collagenase digesting liquid is added dropwise in tissue block surface, with different Sterile ophthalmics
Small scissors is fully shredded to volume about 1-2mm3Size；The tumor tissues collagenase digesting liquid shredded collection is placed on 15ml
In centrifuge tube, a certain amount of digestive juice is added in, and is put into 37 DEG C of CO2It is digested overnight in incubator；Digest overnight sample tissue
And cell suspension centrifugation (300 × g, 7min), supernatant discarding, then centrifuged 3 times with brine；RPMI 1640+10%
The dual anti-precipitations that are resuspended of FBS+ are placed in culture dish, insert 37 DEG C of CO2Incubator culture not move culture dish at least 2 days,
It is adherent beneficial to primary tumor cell；Culture medium is abandoned when cell is grown in ware to 70-80%, PBS is washed 3 times, adds in 1-2ml
EDTA- pancreatin, Microscopic observation when spindle shape fibroblast variation ellipse shape is even circular, add in the new culture containing serum eventually
It only digests, is put into incubator and continues to cultivate.Next day.Whether Microscopic observation also has fibroblast to exist, multiple if any that can carry out
EDTA- pancreatin digests.It builds after being tied to form work(, is respectively designated as Gallbladder cell-Shandong ZJU-0430, people's hilar bile duct cancer cell
It is ZJU-0826 and people hilar bile duct cancerous cell line ZJU-1125, Chinese Typical Representative culture is preserved within 30th in September in 2017
Collection (CCTCC), preservation address are Wuhan Wuhan Universitys of China, and deposit number is respectively：CCTCC NO:
C2017174、CCTCC NO:C2017175 and CCTCC NO:C2017176.In its cellular morphology such as Fig. 1, Fig. 1：A:
ZJU0430；B:ZJU-0826；C:ZJU-1125.
Embodiment 2
Preparation of samples：By 1 × 107It is received after ZJU-0430, ZJU-0826 and ZJU-1125 the cell tryptase enzymic digestion of a culture
It combines in 15ml centrifuge tubes, washing again is collected by centrifugation after cell and is collected by centrifugation；
It is fixed：2.5% glutaraldehyde fixer is added in into cell and fixes 2h, 0.1M phosphate buffers wash 3 times, add in
1%, which starves sour fixer, fixes 2h, and 0.1M phosphate buffers wash 3 times；
Dehydration：Carried out in 4 DEG C of refrigerators respectively 50% ethyl alcohol → → 70% ethyl alcohol → → 90% ethyl alcohol → → 90% ethyl alcohol+
90% acetone (1:1) → → 90% acetone → → 100% acetone (3 times), 15-20min is maintained per procedure；
Embedding：Pure acetone+embedding liquid (2:1) room temperature 3-4h → → pure acetone+embedding liquid (1:2) ambient temperature overnight → → pure bag
Bury 37 DEG C of 2-3h of liquid；
Cure：37 DEG C of oven overnights → → 45 DEG C of baking oven 12h → → 60 DEG C of baking ovens are for 24 hours；
It is prepared by ultra-thin section：50-60nm thickness is cut with ultramicrotome to cut into slices；
Dyeing：3% acetic acid uranium-lead citrate double staining；
Observation film making：It is observed and made film with transmission electron microscope (Hitachi HT7700).
The results are shown in Figure 2, visible abundant organelle (mitochondria, ribosomes and rough surfaced endoplasmic reticulum (RER)), cell chain in endochylema
Visible desmosome between connecing, microfilament, concave nuclear membrane and microvillus sample protrusion in cytoplasm.Fig. 2 is cell transmission electron microscope photo, is schemed
In 2：A,B:ZJU0430；C,D:ZJU-0826；E,F:ZJU-1125.
Embodiment 3
After ZJU-0430, ZJU-0826 and ZJU-1125 cell tryptase enzymic digestion of culture, RPMI 1640+ are resuspended in
It in 10%FBS culture mediums and is counted, final concentration of 2.5 × 10 is obtained after concentration adjusts4/ ml and 5 × 104/ ml's is thin
Born of the same parents' suspension；It is seeded in respectively in 96 orifice plates, per 200 μ l of hole, every group sets 6 multiple holes, and only plus the hole of culture medium is as blank
Control；Culture plate is put into 37 DEG C of incubators and is cultivated, CCK-8 incubations are added in 0,12,24,36,48,60 and 72h time points
Light absorption value is measured at 2.5h, subsequent 450nm.The results are shown in Figure 3, and population doubling time respectively may be about：For 24 hours, for 24 hours and 48h, figure
In 3：A:ZJU0430；B:ZJU-0826；C:ZJU-1125.
Embodiment 4
Need to be carried out at the same time the detection of mycoplasma infection to growing cancer cell, this research is using PCR methods, profit
It, can be simultaneously with the mycoplasma PCR detection kit (HuaAn PCR Mycoplasma Test Kit) of Hangzhou Huaan biotech firm
48 common mycoplasma species are detected, step is as follows：
1. kit forms：
1. 400 μ l of PCR reaction solution (20 μ l/ times, 20 secondary responses)
2. 20 μ l of Taq enzyme (1 μ l/ times, 20 secondary responses)
3. 1000 μ l of DNA release liquids (40 μ l/ times, 25 secondary responses)
4. DNA positive controls 1 are managed (freeze-dried powder)
5. 1000 μ l of atoleine
6. 1 part of operation instructions
2. sample collection and processing：
1. collect cell：People's biliary tract cancerous cell line ZJU-0430, ZJU-0826 and the ZJU-1125 closely converged will be grown to
Epithelial cell is scraped respectively with cell scraper from culture dish, and the culture solution containing cancer cell is moved to respectively in 15ml centrifuge tubes；
2. it centrifuges：More than cell suspension is centrifuged into (12000rpm, 5-l0min)；
3. extract DNA：Supernatant is removed, 40 μ l of DNA release liquids is added in, moves into 2ml EP pipes, fully blow and beat, boil 5min；
4. centrifuging (12000rpm, 5~l0min) again, take supernatant spare as PCR response sample templates.
3. prepare PCR reaction reagents：
1. positive control prepares：DNA positive control powder bottle is first centrifuged into (5000rpm, 5min) on centrifuge, then
It opens bottle cap and adds in 25 μ l deionized waters, fully dissolve, it is spare；
2. prepare PCR reaction solution：PCR reaction solution is taken out from -20 DEG C of refrigerators, 3 repetition detection samples are plus positive right
5 reaction solutions should be prepared altogether according to (positive DNA) and negative control (PBS), take PCR reaction solution l00 μ l, add in 5 μ l Taq enzymes.One
Reaction system needed for secondary property preparation keeps the homogeneity of PCR reactions.
3. abundant mixing PCR reaction solution and Taq enzyme, and dispensed with 21 μ l/ pipes into 5 PCR reaction tubes；
It is right that 4. 4 μ l ready samples (3 repeating pipes), DNA positive controls and PBS feminine genders are respectively added in 5 PCR reaction tubes
According to；
5. PCR reaction tubes are centrifuged into (10000rpm, l0s).
4.PCR reacts and parameter setting：PCR reaction tubes are put into PCR instrument, PCR reactions are carried out by following parameter：
The electrophoresis detection of 5.PCR products：
1. 1 × TAE buffers：L 50 × TAE of ml electrophoretic buffers is taken to add in 49ml deionized waters；
2. prepared by 2.0% Ago-Gel：1.09g agaroses are weighed in conical beaker, add in 50ml l × TAE；
Electrophoretic buffer, micro-wave oven dissolve agarose, slightly 5 μ l GelRed nucleic acid dyes of cold rear addition, pour into gel mold
In, cooled and solidified is spare；
3. loading：Each PCR pipe takes 8 μ l pcr amplification products, adds in the glue hole of agar offset plate, and amplified production includes bromine
Phenol indigo plant indicator；
4. electrophoresis：Electrophoresis is stopped according to bromophenol blue indicator after 110V electrophoresis 20min.
6. gel imaging analysis：Offset plate taking-up after electrophoresis is put into Bio-Rad gel image analysers and observes and claps
According to preservation, it is mycoplasma positive sample the band identical with positive control occur.As shown in Figure 4, with positive and the moon row pair
Photograph ratio, people's biliary tract cancerous cell line ZJU-0430, ZJU-0826 and ZJU-1125 established be not by mycoplasma contamination.
Embodiment 5
Cell prepares：Primary biliary tract cancer cell ZJU-0430, ZJU-0826 and ZJU-1125 of culture are digested with pancreatin
After be suspended in RPMI l640 culture solutions and counted, final concentration of 2.5 × 10 are obtained after concentration adjusts4The cell of/ml
Suspension；
Cell inoculation：The cell suspension of concentrations above is inoculated into respectively in 96 orifice plates, it is (thin containing 5000 per hole 0.2ml
Born of the same parents), every group of 6 multiple holes, only plus the not celliferous hole of culture solution is as blank control；
Chemotherapeutic processing：Suitable chemotherapeutic Gemcitabine is separately added into after cell attachment to be seeded to every group, is set
Various concentration is 10-3μM, 10-2μM, 10-1μM, 1 μM, 10 μM, 102μM, 103μM, it is put into incubator and continues culture for 24 hours；
CCK-8 is detected：Respectively to above every group of cell CCK-8 (1 after specific processing time:10 dilutions, 37 DEG C incubate
Change 2.5h) Activity determination is carried out, and absorbance of the every group of cell in 450nm is recorded, utilize Gradpad Software on Drawing curves.
The results are shown in Figure 5, and the chemotherapeutic Gemcitabine of various concentration can partly inhibit people's hilar bile duct cancerous cell line ZJU-
0826 multiplication, but Gallbladder cell-Shandong ZJU-0430 and people's hilar bile duct cancerous cell line ZJU-1125 are not substantially pressed down
It makes and uses.
Embodiment 6
1. colchicine (final concentration of 0.05mg/ml) is added in into three plants of cancer of bile ducts cell culture fluids of exponential phase of growth
Act on 6 h；
2. being washed after discarding culture solution and using collected by trypsinisation cell, cell is collected in 15ml by PBS washings after centrifuging
Centrifuge bottom of the tube；
3. Hypotonic treatment：Add in hypotonic KCl solution (0.075mol/L) 4- being fully warmed-up in 37 DEG C of water-baths
5ml acts on 20min, abundant turgid cell in 37 DEG C of water-baths；
4. it pre-fixes：Fixer (the methanol of l ml Fresh is directly added into above-mentioned hypotonic medium：Glacial acetic acid, 3：L, V
/ V), gently piping and druming is uniform, centrifuges (1500rpm, 10min) after fixed 5min, abandons supernatant；
5. it fixes again：The fixer of 6-8ml Fresh is added in, room temperature fixes 30min；
6. blow piece：5-6 drop fixers will be added after more than specimen centrifuge again, examination takes 1 drop, drops on 4 DEG C of distilled water slides,
It is toasted rapidly on alcolhol burner flame and blows piece；
7. Giemsa is dyed：Appropriate Giemsa working solutions dyeing 10-15min is added dropwise in the above surface of glass slide blown, from
Naturally dry obtains metaphase chromosome sample after water rinses slide.
Fig. 6 is ZJU-0430, ZJU-0826, the representative single celled karyotyping of ZJU-1125 cells.(A)ZJU-0430
45th generation unicellular representative caryogram；(B) the 67th generations of ZJU-0826 unicellular representative caryogram；(C) ZJU-1125 the 87th
For unicellular representative caryogram.Caryogram collection of illustrative plates shows that the three plants of cells established in chromosome number and structure occur
Abnormal, chromosome number is hypo-triploid caryogram, is distributed in mostly between 65-80, textural anomaly include chromosome increase, missing and
Transposition etc..
Embodiment 7
1.ZJU-0430 ZJU-0826 and ZJU-1125 cell samples prepare：
A. it will be suspended in PBS and carry out after ZJU-0430, ZJU-0826 and ZJU-1125 the cell tryptase enzymic digestion of culture
It counts, final concentration of 2 × 10 is obtained after concentration adjusts6The cell suspension of/ml；
B. above-mentioned cell suspension 0.2ml is taken, is added in the circle of FTA specimen collecting cards, is air-dried in super-clean bench；
C. the FTA card samples of 1.2mm diameters are taken in circle center with Harris card punch, it is spare.
2. it uses18D kits carry out DNA cloning：
1mlD 5×Master Mix
1ml18D 5×Primer Pair Mix
25μl 2800M Control DNA,10ng/μl
5×1,250μl Water,Amplification Grade
A. kit component (ecru lid) after expanding：
50μl18D Allelic Ladder Mix
2×150μl CC5Internal Lane Standard 500
B. according to the form below configures in 96 hole reaction plates of MicroAmp18D systems expand mixed liquor；
C. the FTA card samples of spare 1.2mm diameters are added in 96 hole reaction plates in sample aperture；
D., the positive control and negative control of amplification are set：2800M Control DNA are diluted to 5ng/ μ l, take 1 μ l
DNA dilutions be added in the reacting hole containing 25 μ l pcr amplification reaction liquid as positive control, make 1.2mm diameters
Blank FTA cards as negative control, can be used to detect perforating device has no cross contamination.
E. existFollowing amplification thermal circulation parameters are set on 9700 thermal cyclers of PCR system：
F. ABI is used3500 × 1 type genetic analyzers test and analyze amplified fragments；
G. existFragment data in 1D-X analysis softwares in steps for importing 3 is analyzed；
H. by more than analysis result E-mail to ATCC (STRtesting@atcc.org) carry out identification and analysis；
I.ATCC provides Species estimation examining report.
As shown in fig. 7, STR testing results confirm newly-established three plants of biliary tract cancerous cell lines：Gallbladder cell-Shandong ZJU-
0430, people hilar bile duct cancerous cell line ZJU-0826 and people hilar bile duct cancerous cell line ZJU-1125, with ATCC and DSMZ
Cytogenetics information in the foremost culture collection mechanism in two whole world is compared, and discovery is not affected by the dirt of other cells
Dye.ATCC number:STRA6989, STRA6988, STRA6986；In Fig. 7：A-D:ZJU-0430；E-H:ZJU-0826；I-
L: ZJU-1125。
Embodiment 8
1. cell prepares：After (800rpm, 5min) is centrifuged after three plants of cancer of bile ducts cell tryptase enzymic digestions of exponential phase of growth
Centrifugation 3 times is washed with serum free medium；
2. cell is resuspended in PBS buffer solution, it is 5 × 10 to count adjustment cell density7/ml；
3. the above cell suspension of 0.2ml (is contained about 1 × 107A cell) it is injected into the BABL/c mouse backs of 4-6 weeks
Subcutaneously (n=3), Mouse feeder is in SPF grades of laminar flow；
4. observation 4 weeks palpation and measures Tumor diameter size weekly；
Mouse is euthanized after 5.4 weeks, knurl bulk measurement is taken out and takes pictures, part knurl body is taken to be soaked in 10% formalin molten
Liquid carries out routine paraffin wax embedding and microsection manufacture, spare.
As shown in figure 8, ZJU-0430 and ZJU-1125 on BABL/C nude mices into knurl, and prolonging with inoculation time
Long, transplantable tumor tumor volume gradually increases, and pathologic finding shows to be gland cancer, and ZJU-0826 is into knurl, in Fig. 8：A:ZJU-
0430,B: ZJU-1125。
Above-mentioned specific embodiment is used for illustrating the present invention, is merely a preferred embodiment of the present invention rather than to this
Invention is limited, and in the protection domain of spirit and claims of the present invention, to any modification of the invention made, is equal
Replace, improve etc., both fall within protection scope of the present invention.
Claims (9)
1. people's biliary tract cancerous cell line, which is characterized in that people's biliary tract cancerous cell line includes：It is respectively designated as Human gallbladder carcinoma cell
It is ZJU-0430, people hilar bile duct cancerous cell line ZJU-0826 and people hilar bile duct cancerous cell line ZJU-1125, and preservation
In China typical culture collection center, deposit number is respectively：CCTCC NO: C2017174、CCTCC NO: C2017175
With CCTCC NO:People's biliary tract cancerous cell line of C2017176.
2. people's biliary tract cancerous cell line according to claim 1, which is characterized in that people's biliary tract cancerous cell line further includes：
It is respectively designated as Gallbladder cell-Shandong ZJU-0430, people hilar bile duct cancerous cell line ZJU-0826 and people's hilar cholangiocarcinoma
Cell line ZJU-1125, and China typical culture collection center is preserved in, deposit number is respectively：CCTCC NO:
C2017174、CCTCC NO:C2017175 and CCTCC NO:The daughter cell system of people's biliary tract cancerous cell line of C2017176.
3. people's biliary tract cancerous cell line according to claim 1 or 2, which is characterized in that people's biliary tract cancerous cell line is in body
It is typical polygon epithelioid cell during outer culture, under light microscopic, there is typical Malignant Epithelium feature, cell multiplication under Electronic Speculum
Time respectively may be about for 24 hours, for 24 hours, 48h.
4. people's biliary tract cancerous cell line according to claim 1 or 2, which is characterized in that people's biliary tract cancerous cell line is in courage
Road cancer Clinicopathologic Diagnosis is gland cancer；Karyotyping the results show its occur in chromosome number and structure it is abnormal.
5. people's biliary tract cancerous cell line according to claim 1 or 2, which is characterized in that chemotherapeutic gemcitabine can partly press down
People's hilar bile duct cancerous cell line ZJU-0826 processed, but it is thin to Gallbladder cell-Shandong ZJU-0430 and people's hilar cholangiocarcinoma
Born of the same parents system ZJU-1125 does not have any inhibitory action.
6. people's biliary tract cancerous cell line according to claim 1 or 2, which is characterized in that people's biliary tract cancerous cell line is propping up
When Mycoplasma Detection Reagent box detects, PCR results are feminine gender, are not affected by mycoplasma contamination；And it is deposited through American Type Culture collection
The biliary tract cancerous cell line that storehouse short tandem repeat identification finds to establish derives from people, and not by other cell contaminations.
7. a kind of people's biliary tract cancerous cell line according to claim 1 or claim 2 is in the cell model of cancer of bile ducts study of incident mechanism
Application.
8. a kind of purposes of people's biliary tract cancerous cell line, which is characterized in that people's biliary tract cell line according to claim 1 or claim 2 is used
In the model for preparing the generation cancer of bile ducts in immune deficiency mammal, wherein ZJU-0328 and ZJU-1125 tumor formation rates are
100%, ZJU-0826 be not then into knurl.
9. the purposes of people's biliary tract cancerous cell line according to claim 8, which is characterized in that the immune deficiency mammal
For immune deficiency nude mouse.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810004140.4A CN108048401A (en) | 2018-01-03 | 2018-01-03 | People's biliary tract cancerous cell line and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810004140.4A CN108048401A (en) | 2018-01-03 | 2018-01-03 | People's biliary tract cancerous cell line and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108048401A true CN108048401A (en) | 2018-05-18 |
Family
ID=62126161
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810004140.4A Pending CN108048401A (en) | 2018-01-03 | 2018-01-03 | People's biliary tract cancerous cell line and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108048401A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111117965A (en) * | 2020-01-08 | 2020-05-08 | 宁波市医疗中心李惠利医院 | Rapid purification method of high-purity primary tumor cells |
| CN117004571A (en) * | 2023-07-29 | 2023-11-07 | 兰州大学第一医院 | Human hilar bile duct carcinoma sarcoma cell line CBC2T-2 and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999043196A2 (en) * | 1998-02-25 | 1999-09-02 | Sumitomo Electric Industries, Ltd. | Method of cancer diagnosis based on the detection of chromosome abnormality |
| CN104053438A (en) * | 2011-10-18 | 2014-09-17 | 味之素株式会社 | Therapeutic agent for pancreatic cancer and/or biliary tract cancer |
| CN105255832A (en) * | 2014-07-17 | 2016-01-20 | 上海立迪生物技术有限公司 | Human bile duct cancer cell line and applications thereof |
| CN106110312A (en) * | 2016-07-14 | 2016-11-16 | 广东天普生化医药股份有限公司 | Ulinastatin purposes in preparation treatment carcinoma of gallbladder medicine |
| CN107412768A (en) * | 2017-06-13 | 2017-12-01 | 南京大学 | GGPPS suppression is preparing the purposes in treating obstructive jaundice liver injury medicament |
-
2018
- 2018-01-03 CN CN201810004140.4A patent/CN108048401A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999043196A2 (en) * | 1998-02-25 | 1999-09-02 | Sumitomo Electric Industries, Ltd. | Method of cancer diagnosis based on the detection of chromosome abnormality |
| CN104053438A (en) * | 2011-10-18 | 2014-09-17 | 味之素株式会社 | Therapeutic agent for pancreatic cancer and/or biliary tract cancer |
| CN105255832A (en) * | 2014-07-17 | 2016-01-20 | 上海立迪生物技术有限公司 | Human bile duct cancer cell line and applications thereof |
| CN106110312A (en) * | 2016-07-14 | 2016-11-16 | 广东天普生化医药股份有限公司 | Ulinastatin purposes in preparation treatment carcinoma of gallbladder medicine |
| CN107412768A (en) * | 2017-06-13 | 2017-12-01 | 南京大学 | GGPPS suppression is preparing the purposes in treating obstructive jaundice liver injury medicament |
Non-Patent Citations (7)
| Title |
|---|
| HIROSHI EGAMI等: "ESTABLISHMENT OF A CELL LINE OF GALLBLADDER CARCINOMA (GBK-1)PRODUCING HUMAN COLONY STIMULATING FACTOR", 《JPN. J. CANCER RES. (GANN)》 * |
| SHOHEI KOYAMA等: "ESTABLISHMENT OF A CELL LINE (G-415) FROM A HUMAN GALLBLADDER CARCINOMA", 《GANN》 * |
| 刘博等: "人胆囊癌细胞系GBC-SD的建立", 《山东医科大学学报》 * |
| 吴小鹏等: "肝门部胆管癌细胞系（FRH-0201）的建立及生物学特性研究", 《中华医学杂志》 * |
| 国家药典委员会编: "《中华人民共和国药典 2010年版 第二增补本》", 30 September 2013, 中国医药科技出版社 * |
| 张志发等: "人胆囊癌细胞系GBC—SD中侧群细胞的耐药性研究", 《中华肝胆外科杂志》 * |
| 杨耀琴等: "人原发性胆囊癌细胞系SGC-996的建立", 《同济大学学报(医学版)》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111117965A (en) * | 2020-01-08 | 2020-05-08 | 宁波市医疗中心李惠利医院 | Rapid purification method of high-purity primary tumor cells |
| CN117004571A (en) * | 2023-07-29 | 2023-11-07 | 兰州大学第一医院 | Human hilar bile duct carcinoma sarcoma cell line CBC2T-2 and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108034636B (en) | Human breast cancer cell line and application | |
| CN109554346A (en) | A kind of lung cancer organoid model and its application in tumor research | |
| CN114292816B (en) | Lung cancer organoid culture solution, and culture reagent combination and culture method thereof | |
| Dexter et al. | Establishment and characterization of two human pancreatic cancer cell lines tumorigenic in athymic mice | |
| Esparza-López et al. | Deriving primary cancer cell cultures for personalized therapy | |
| CN102719403B (en) | A kind of human pancreas's adenosquamous carcinoma clone and establishment method thereof and application | |
| CN108611322A (en) | Method for building up and the application of breast cancer circulating tumor cell system CTC-3, culture medium and CTC-3 | |
| CN102067828B (en) | Highly metastatic model of human melanoma, cell subline, creation methods, and dynamic detection of metastasis | |
| CN114717191A (en) | Human intrahepatic bile duct cancer cell line ICC-X3 and application thereof | |
| CN108048401A (en) | People's biliary tract cancerous cell line and application | |
| CN106367393A (en) | Mouse prostate cancer circulating tumor cell line and prostate cancer circulating tumor cell isolating and culturing method | |
| Preksha et al. | Cell culture techniques in gastrointestinal research: Methods, possibilities and challenges | |
| CN109355261B (en) | Urinary bladder cancer cell culture medium and urinary bladder cancer cell in-vitro culture method | |
| CN114908039A (en) | Culture medium for stomach cancer organoid and culture method without bracket thereof | |
| CN106940372A (en) | Detect methylene blue, upper conversion nano grain modification fibrosis paper sensor of Telomerase and preparation method thereof | |
| CN114606194A (en) | Human intrahepatic bile duct cancer cell line ICC-X1 and application thereof | |
| CN1879892B (en) | Method for establishing human extranodal rhinoid NK/T cell lymphoma animal model | |
| CN105255832A (en) | Human bile duct cancer cell line and applications thereof | |
| CN106479981B (en) | A kind of people's Endometrial carcinoma cell line and its method for building up | |
| CN115786262B (en) | Human hilar bile duct cancer cell line CBC3T-1 and application thereof | |
| CN105039569A (en) | Method for analyzing breakpoints of reciprocal translocation chromosomes | |
| CN108070567A (en) | A kind of urine derived cell strain of immortalization and its construction method | |
| CN105062973B (en) | One plant carries HPV feminine gender penis squamous cell carcinomas system that TP53 is mutated and application thereof | |
| CN108467855A (en) | New lung specificity transfer liver cancer cells and its preparation | |
| CN105861442B (en) | Height transfer hepatoma cell strain and its construction method and application |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180518 |
|
| RJ01 | Rejection of invention patent application after publication |