CN113025576B - 一种人源性腺激素型垂体腺瘤干细胞细胞系的构建方法及其应用 - Google Patents
一种人源性腺激素型垂体腺瘤干细胞细胞系的构建方法及其应用 Download PDFInfo
- Publication number
- CN113025576B CN113025576B CN202110272526.5A CN202110272526A CN113025576B CN 113025576 B CN113025576 B CN 113025576B CN 202110272526 A CN202110272526 A CN 202110272526A CN 113025576 B CN113025576 B CN 113025576B
- Authority
- CN
- China
- Prior art keywords
- pituitary adenoma
- cell line
- stem cell
- tumor
- human
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 208000007913 Pituitary Neoplasms Diseases 0.000 title claims abstract description 110
- 210000000130 stem cell Anatomy 0.000 title claims abstract description 90
- 201000005746 Pituitary adenoma Diseases 0.000 title claims abstract description 84
- 206010061538 Pituitary tumour benign Diseases 0.000 title claims abstract description 84
- 208000021310 pituitary gland adenoma Diseases 0.000 title claims abstract description 84
- 238000010276 construction Methods 0.000 title abstract description 9
- 210000004027 cell Anatomy 0.000 claims abstract description 42
- 239000003814 drug Substances 0.000 claims abstract description 20
- 238000011160 research Methods 0.000 claims abstract description 15
- 238000012216 screening Methods 0.000 claims abstract description 10
- 238000000338 in vitro Methods 0.000 claims abstract description 9
- 230000008261 resistance mechanism Effects 0.000 claims abstract description 5
- 239000000745 gonadal hormone Substances 0.000 claims description 28
- 238000000034 method Methods 0.000 claims description 25
- 229940079593 drug Drugs 0.000 claims description 16
- 238000010171 animal model Methods 0.000 claims description 13
- 230000002710 gonadal effect Effects 0.000 claims description 13
- 241000321096 Adenoides Species 0.000 claims description 11
- 210000002534 adenoid Anatomy 0.000 claims description 11
- 239000000126 substance Substances 0.000 claims description 8
- 238000004321 preservation Methods 0.000 claims description 6
- 238000012360 testing method Methods 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 4
- 230000009545 invasion Effects 0.000 claims description 3
- 238000009629 microbiological culture Methods 0.000 claims description 3
- 230000006907 apoptotic process Effects 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 230000008506 pathogenesis Effects 0.000 claims description 2
- 230000035755 proliferation Effects 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims 1
- 210000004881 tumor cell Anatomy 0.000 abstract description 20
- 239000005556 hormone Substances 0.000 abstract description 11
- 229940088597 hormone Drugs 0.000 abstract description 11
- 210000004907 gland Anatomy 0.000 abstract description 5
- 239000000463 material Substances 0.000 abstract description 5
- 206010059866 Drug resistance Diseases 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 4
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 3
- 238000012827 research and development Methods 0.000 abstract description 3
- 206010028980 Neoplasm Diseases 0.000 description 50
- 210000001519 tissue Anatomy 0.000 description 13
- 230000004069 differentiation Effects 0.000 description 11
- 238000002474 experimental method Methods 0.000 description 10
- 210000002966 serum Anatomy 0.000 description 9
- 239000006285 cell suspension Substances 0.000 description 8
- 239000002771 cell marker Substances 0.000 description 7
- 238000010790 dilution Methods 0.000 description 7
- 239000012895 dilution Substances 0.000 description 7
- 238000011161 development Methods 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 230000028327 secretion Effects 0.000 description 6
- 101000610551 Homo sapiens Prominin-1 Proteins 0.000 description 5
- 102000008730 Nestin Human genes 0.000 description 5
- 108010088225 Nestin Proteins 0.000 description 5
- 210000003743 erythrocyte Anatomy 0.000 description 5
- 238000003125 immunofluorescent labeling Methods 0.000 description 5
- 210000005055 nestin Anatomy 0.000 description 5
- 208000010916 pituitary tumor Diseases 0.000 description 5
- 229960005322 streptomycin Drugs 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000283707 Capra Species 0.000 description 4
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 4
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 4
- 101710126211 POU domain, class 5, transcription factor 1 Proteins 0.000 description 4
- 101100247004 Rattus norvegicus Qsox1 gene Proteins 0.000 description 4
- 239000006143 cell culture medium Substances 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000012091 fetal bovine serum Substances 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 239000003163 gonadal steroid hormone Substances 0.000 description 4
- 238000010166 immunofluorescence Methods 0.000 description 4
- 230000007246 mechanism Effects 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 102000006771 Gonadotropins Human genes 0.000 description 3
- 108010086677 Gonadotropins Proteins 0.000 description 3
- 108010076089 accutase Proteins 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 230000001079 digestive effect Effects 0.000 description 3
- 238000007710 freezing Methods 0.000 description 3
- 230000008014 freezing Effects 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
- 239000002622 gonadotropin Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000004017 serum-free culture medium Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000010257 thawing Methods 0.000 description 3
- 102000018997 Growth Hormone Human genes 0.000 description 2
- 108010051696 Growth Hormone Proteins 0.000 description 2
- 102000003946 Prolactin Human genes 0.000 description 2
- 108010057464 Prolactin Proteins 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 238000001647 drug administration Methods 0.000 description 2
- 239000000122 growth hormone Substances 0.000 description 2
- 238000003364 immunohistochemistry Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000007917 intracranial administration Methods 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 210000001328 optic nerve Anatomy 0.000 description 2
- 230000002980 postoperative effect Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 229940097325 prolactin Drugs 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000002271 resection Methods 0.000 description 2
- 239000012679 serum free medium Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 238000010254 subcutaneous injection Methods 0.000 description 2
- 239000007929 subcutaneous injection Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000005740 tumor formation Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 208000003200 Adenoma Diseases 0.000 description 1
- 206010001233 Adenoma benign Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 101100298998 Caenorhabditis elegans pbs-3 gene Proteins 0.000 description 1
- 239000000055 Corticotropin-Releasing Hormone Substances 0.000 description 1
- 102000012673 Follicle Stimulating Hormone Human genes 0.000 description 1
- 108010079345 Follicle Stimulating Hormone Proteins 0.000 description 1
- 206010018338 Glioma Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108700011259 MicroRNAs Proteins 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 235000012904 Prunus salicina Nutrition 0.000 description 1
- 240000005049 Prunus salicina Species 0.000 description 1
- 235000003681 Prunus ussuriensis Nutrition 0.000 description 1
- 101150037203 Sox2 gene Proteins 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 102000011923 Thyrotropin Human genes 0.000 description 1
- 108010061174 Thyrotropin Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 230000003023 adrenocorticotropic effect Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 210000004004 carotid artery internal Anatomy 0.000 description 1
- 210000000711 cavernous sinus Anatomy 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- IDLFZVILOHSSID-OVLDLUHVSA-N corticotropin Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)NC(=O)[C@@H](N)CO)C1=CC=C(O)C=C1 IDLFZVILOHSSID-OVLDLUHVSA-N 0.000 description 1
- 229960000258 corticotropin Drugs 0.000 description 1
- 238000007428 craniotomy Methods 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000029578 entry into host Effects 0.000 description 1
- 238000011124 ex vivo culture Methods 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 229940028334 follicle stimulating hormone Drugs 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000002055 immunohistochemical effect Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 230000002601 intratumoral effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 206010027191 meningioma Diseases 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 210000003928 nasal cavity Anatomy 0.000 description 1
- 210000001989 nasopharynx Anatomy 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 239000000101 novel biomarker Substances 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 210000003903 pelvic floor Anatomy 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 210000003718 sphenoid sinus Anatomy 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000007910 systemic administration Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0693—Tumour cells; Cancer cells
- C12N5/0695—Stem cells; Progenitor cells; Precursor 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
- A01K67/0271—Chimeric vertebrates, e.g. comprising exogenous cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
-
- 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
- C12N5/0613—Cells from endocrine organs
- C12N5/0616—Pituitary gland
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/90—Serum-free medium, which may still contain naturally-sourced components
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/11—Epidermal growth factor [EGF]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2503/00—Use of cells in diagnostics
- C12N2503/02—Drug screening
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
- C12N2509/10—Mechanical dissociation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2500/00—Screening for compounds of potential therapeutic value
- G01N2500/10—Screening for compounds of potential therapeutic value involving cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Endocrinology (AREA)
- Urology & Nephrology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Hematology (AREA)
- Environmental Sciences (AREA)
- Pathology (AREA)
- Animal Behavior & Ethology (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Physics & Mathematics (AREA)
- Biodiversity & Conservation Biology (AREA)
- Developmental Biology & Embryology (AREA)
- Oncology (AREA)
- Animal Husbandry (AREA)
- General Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Diabetes (AREA)
- Food Science & Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Rheumatology (AREA)
- Epidemiology (AREA)
- Public Health (AREA)
Abstract
本发明公开了一种人源性腺激素型垂体腺瘤干细胞细胞系的构建方法及其应用,通过前期探索及优化培养条件,成功建立了来源于人的性腺激素型垂体腺瘤干细胞细胞系,该细胞系具有稳定的生物学特性、高体外成球能力、干细胞特性,可稳定多次传代,对其进行分化可产生分化后的肿瘤细胞,并且可持续自我更新,为针对性腺激素型垂体腺瘤的基础科研、药物的研发与筛选、耐药机理的研究、以及诊断试剂的研发提供了实验材料,为临床上进一步提高性腺激素型垂体腺瘤的治疗效果奠定了基础。
Description
技术领域
本发明属于生物技术领域,具体而言,涉及一种人源性腺激素型垂体腺瘤干细胞细胞系的构建方法及其应用。
背景技术
垂体腺瘤(Pituitary adenomas)是一种常见的颅内肿瘤,发病率仅次于胶质瘤和脑膜瘤,约占颅内肿瘤的15%(Wu S,Gu Y,Huang Y,Wong TC,Ding H,Liu T, Zhang Y,Zhang X.Novel Biomarkers for Non-functioning Invasive Pituitary Adenomas wereIdentified by Using Analysis of microRNAs Expression Profile[J]. BiochemGenet.2017,55(3):253-267.),手术切除是垂体腺瘤的首选治疗方式。经过近一个世纪的发展,肿瘤的全切率、术后患者生活质量都得到了显著性的提高和保证。然而,临床上约有25%-55%的垂体腺瘤病例具有侵袭性生长的生物学特征,即易包裹颈内动脉、视神经等重要结构,侵入海绵窦,甚至突破鞍底侵入蝶窦、鼻腔及鼻咽部,给手术造成极大困难,也无法达到全切程度。因此,虽然绝大多数垂体腺瘤为良性病变,但却能够长期困扰患者。
根据肿瘤是否具有分泌功能,垂体腺瘤可分为无分泌功能性垂体腺瘤和有分泌功能性垂体腺瘤,无分泌功能性垂体腺瘤约占垂体腺瘤的30%,可不改变激素分泌水平,主要临床表现为肿瘤的占位效应,如压迫视神经造成视力视野改变等,有分泌功能性垂体腺瘤约占垂体腺瘤的70%,主要包括:泌乳素型垂体腺瘤(PRL)、生长激素型垂体腺瘤(GH)、促肾上腺皮质激素型垂体腺瘤(ACTH)、促甲状腺激素型垂体腺瘤(TSH)、性腺激素型(卵泡刺激素型,FSH型)垂体腺瘤。本发明人通过前期的回顾性研究发现,外周血无激素改变的垂体腺瘤患者中,术后病理为性腺激素型垂体腺瘤的患者占一定的比例,在临床上多表现为静默型,即病理染色FSH及转录因子SF-1阳性。
肿瘤干细胞是肿瘤组织中一小部分具有干细胞特性的细胞。肿瘤生长的微环境下,干细胞可进行自我更新并分化为肿瘤细胞。随着肿瘤干细胞理论的建立与发展,为建立合适的、贴近于真实病例的肿瘤模型及研究肿瘤发生发展机制提供了有力的理论支撑。目前,相对于生长激素型、无功能型和泌乳素型垂体腺瘤,性腺激素型垂体腺瘤为一种相对少见的亚型,目前临床上和基础研究中对这一类型的垂体腺瘤认识不足,未见特殊药物对其进行精准治疗,而针对性腺激素型垂体腺瘤的应用基础研究中缺乏较为成熟的人源性腺激素型垂体腺瘤肿瘤干细胞系。本发明针对目前本领域存在的这一问题,通过前期探索及优化培养条件,成功建立了来源于人的性腺激素型垂体腺瘤干细胞细胞系,对其进行分化可产生分化后的肿瘤细胞,并且可持续自我更新,为下一步探索垂体腺瘤的发生发展及治疗过程提供了坚实的实验方法支持。
发明内容
为了弥补现有技术关于人源性腺激素型垂体腺瘤肿瘤干细胞系的空白,本发明的目的在于提供一种人源性腺激素型垂体腺瘤干细胞细胞系的构建方法及其应用,通过该方法构建得到的人源性腺激素型垂体腺瘤干细胞细胞系填补了现有技术的空白,为针对性腺激素型垂体腺瘤的基础科研、药物的研发与筛选、耐药机理的研究、以及诊断试剂的研发提供了实验材料,为探索性腺激素型垂体腺瘤的发生发展机制提供了坚实的实验方法支持,并且为临床上进一步提高性腺激素型垂体腺瘤的治疗效果奠定了基础。
为了解决上述技术问题,本发明采用了如下技术方案:
本发明的第一方面提供了一种人源性腺激素型垂体腺瘤干细胞细胞系,其保藏在中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC No.21018。
本发明的第二方面提供了如本发明第一方面所述的人源性腺激素型垂体腺瘤干细胞细胞系的子代细胞。
进一步,所述子代细胞经本发明第一方面所述的人源性腺激素型垂体腺瘤干细胞细胞系培养后获得。
本发明的第三方面提供了本发明第一方面所述的人源性腺激素型垂体腺瘤干细胞细胞系的构建方法。
进一步,所述方法包括如下步骤:
(1)将肿瘤组织制备成肿瘤单细胞悬液;
(2)将步骤(1)中所述的单细胞悬液置于无血清培养基中进行培养,形成稳定的性腺激素型垂体腺瘤干细胞细胞系。
进一步,步骤(1)中所述的肿瘤组织为性腺激素型垂体腺瘤组织;
优选地,所述性腺激素型垂体腺瘤组织来源于人。
进一步,步骤(2)中所述的无血清培养基为添加EGF、bFGF、N2、B27、青- 链霉素的无血清培养基;
优选地,所述EGF的浓度为20ng/mL;
优选地,所述bFGF的浓度为20ng/mL;
优选地,所述青-链霉素的浓度为200U/mL。
进一步,步骤(2)中所述的培养的时间为3周以上,每2-3天换液一次,每7 天传一代。
进一步,步骤(1)中所述的肿瘤单细胞悬液的制备可使用本领域技术人员已知的任何常规方法和手段进行,其中,所述常规方法和手段包括(但不限于):物理方法、酶解消化法、物理-酶解消化法。
在本发明的实施例中,步骤(1)中所述的制备肿瘤单细胞悬液的方法优选为物理-酶解消化法。
进一步,所述的制备肿瘤单细胞悬液的方法具体包括如下步骤:将性腺激素型垂体腺瘤患者的垂体腺瘤肿瘤组织标本离体后迅速放入无菌的无血清培养基中,通过物理机械切割的方式将肿瘤组织切割成细小的碎片,然后应用消化液进一步将切割后的细小的碎片消化为单个细胞,再使用红细胞裂解液充分去除红细胞后,通过40μm的筛网,以去除大块的未充分消化的碎片,制备得到肿瘤单细胞悬液;
优选地,所述消化液为accutase细胞消化液。
根据本发明的构建方法获得的人源性腺激素型垂体腺瘤干细胞细胞系可使用本领域常规方法进行冻存,例如,将构建得到的人源性腺激素型垂体腺瘤干细胞细胞系离心后重悬于细胞冻存液中,滴度冻存盒内-80℃静置过夜,然后迅速移入液氮中进行冻存。
复苏冻存的根据本发明的方法获得的细胞的方法是本领域技术人员已知或者可以容易确定的复苏冻存细胞的方法,例如,从液氮中取出冻存细胞,置37-40℃水浴中快速振摇使细胞快速融化,用新鲜的培养液悬浮细胞后,1000rpm离心 5min,细胞用适当体积的培养基悬起,进行传代培养。
进一步,本发明还提供了对构建得到的人源性腺激素型垂体腺瘤干细胞细胞系进行鉴定。
进一步所述鉴定包括(但不限于):干细胞成球实验鉴定细胞的成球能力,免疫荧光染色法鉴定肿瘤干细胞标志物的表达情况,血清分化实验鉴定细胞是否具有分化潜能,对分化后的肿瘤细胞进行免疫组化鉴定肿瘤干细胞标志物。
在本发明的实施例中,所述干细胞成球实验鉴定细胞的成球能力包括对离体培养18天后的人源性腺激素型垂体腺瘤干细胞细胞系进行鉴定,证明了本发明所述的人源性腺激素型垂体腺瘤干细胞细胞系具有高的体外成球能力。
在本发明的实施例中,所述免疫荧光染色法鉴定肿瘤干细胞标志物的表达情况包括对本发明所述的人源性腺激素型垂体腺瘤干细胞细胞系中Sox2、Oct4、 Nestin、CD133的表达情况进行鉴定,证明了本发明所述的人源性腺激素型垂体腺瘤干细胞细胞系具有干细胞的特性。
在本发明的实施例中,所述血清分化实验鉴定细胞是否具有分化潜能包括通过在培养基中添加15%FBS诱导肿瘤干细胞的分化,对同期对照培养的肿瘤干细胞及分化细胞拍照计数,证明了本发明所述的人源性腺激素型垂体腺瘤干细胞细胞系具有分化潜能。
在本发明的实施例中,所述对分化后的肿瘤细胞进行免疫组化鉴定肿瘤干细胞标志物包括对分化后的肿瘤细胞中Sox2、Oct4、Nestin、CD133的表达情况进行鉴定,证明了由本发明所述的人源性腺激素型垂体腺瘤干细胞分化得到的肿瘤细胞的干性明显降低。
本发明的第四方面提供了本发明第一方面所述的细胞系在如下任一方面所述的应用:
(1)在制备性腺激素型垂体腺瘤细胞模型中的应用;
(2)在制备干细胞异种移植的性腺激素型垂体腺瘤动物模型中的应用;
(3)在筛选预防和/或治疗性腺激素型垂体腺瘤的药物中的应用;
(4)在建立性腺激素型垂体腺瘤发病机制、侵袭机制的研究平台中的应用;
(5)在建立耐药机制研究平台中的应用。
进一步,所述本发明第一方面提供的细胞系在制备干细胞异种移植的性腺激素型垂体腺瘤动物模型中的应用包括将本发明第一方面所述的干细胞细胞系异种移植到非人哺乳动物的体内,进而形成干细胞异种移植的性腺激素型垂体腺瘤动物模型;
优选地,所述非人哺乳动物包括小鼠、大鼠;
更优选地,所述非人哺乳动物为小鼠;
最优选地,所述非人哺乳动物为免疫缺陷的小鼠。
进一步,所述干细胞异种移植的性腺激素型垂体腺瘤动物模型包括(但不限于):皮下成瘤模型、原位肿瘤模型、转移肿瘤模型。
进一步,所述皮下成瘤模型采用的移植接种方式是单细胞悬液皮下注射。
进一步,在筛选预防和/或治疗性腺激素型垂体腺瘤的药物中的应用包括将不同药物添加到本发明第一方面所述的干细胞细胞系培养基中,观察细胞状态的变化情况,以筛选得到初步有效的候选药物,然后将候选药物施用于本发明所述的性腺激素型垂体腺瘤细胞模型和/或性腺激素型垂体腺瘤动物模型中,观察并比较施用药物组和未施用药物组的细胞和/或动物的存活状态和/或肿瘤的大小、存活期等,筛选能够抑制细胞存活和/或抑制动物模型中肿瘤的生长、延长动物模型存活期的药物,将筛选出的药物作为潜在的预防和/或治疗性腺激素型垂体腺瘤的药物。
本发明的第五方面提供了一种用于治疗性腺激素型垂体腺瘤的候选药物的筛选方法。
进一步,所述方法包括如下步骤:
(1)将待测物质施用于细胞系模型中;
(2)若步骤(1)中所述的待测物质施用后,导致细胞凋亡或抑制细胞增殖,则所述待测物质为治疗性腺激素型垂体腺瘤的候选药物。
进一步,所述步骤(1)中的细胞系模型如本发明第一方面所述的细胞系。
本发明的第六方面提供了一种用于治疗性腺激素型垂体腺瘤的候选药物的筛选方法,
进一步,所述方法包括如下步骤:
(1)将待测物质施用于动物模型中;
(2)若步骤(1)中所述的待测物质施用后,导致动物模型性腺激素型垂体腺瘤症状改善或治愈,则所述待测物质为治疗性腺激素型垂体腺瘤的候选药物;
优选地,所述的动物模型是采用本发明第一方面所述的干细胞细胞系经干细胞异种移植得到的性腺激素型垂体腺瘤动物模型。
进一步,所述将待测物质施用于动物模型的施用方法采用的是本领域常规的施用方法。
进一步,所述本领域常规的施用方法包括(但不限于):瘤内给药、肌肉内给药、腹腔注射给药、静脉注射给药、皮下注射给药、尾静脉注射给药、皮肤局部给药、全身给药、灌胃、口服中的一种或几种方法联合应用。
生物材料样品的保藏信息:
分类命名:人源性腺激素型垂体腺瘤干细胞细胞系
保藏单位:中国微生物菌种保藏管理委员会普通微生物中心;
地址:北京市朝阳区北辰西路1号院3号;
保藏日期:2020年12月02日;
保藏编号:CGMCC No.21018;
分类命名:人源静默型促性腺激素型垂体腺瘤干细胞细胞系。
本发明的优点和有益效果如下:
1、本发明首次构建了人源性腺激素型垂体腺瘤干细胞细胞系,填补了目前这一领域的技术空白,且该细胞系具有稳定的生物学特性、高体外成球能力、干细胞特性,可稳定多次传代,对其分化可产生分化后的肿瘤细胞,并且可持续自我更新。
2、本发明提供的人源性腺激素型垂体腺瘤干细胞细胞系为针对性腺激素型垂体腺瘤的基础科研、药物的研发与筛选、耐药机理的研究、以及诊断试剂的研发提供了实验材料,为临床上进一步提高性腺激素型垂体腺瘤的治疗效果奠定了基础。
附图说明
以下,结合附图来详细说明本发明的实施方案,其中:
图1显示的是离体培养18天后的性腺激素型垂体腺瘤干细胞在光镜下的结果图;
图2显示的是免疫荧光染色法对性腺激素型垂体腺瘤干细胞中肿瘤干细胞标志物鉴定的结果图;
图3显示的是对添加血清分化一周后的肿瘤细胞及其对照的肿瘤干细胞分别拍照得到的结果图,其中,A图:对照的肿瘤干细胞,B图:肿瘤细胞;
图4显示的是对分化的肿瘤细胞进行肿瘤干细胞标志物鉴定的结果图。
具体实施方式
下面结合具体实施例,进一步阐述本发明,仅用于解释本发明,而不能理解为对本发明的限制。本领域的普通技术人员可以理解为:在不脱离本发明的原理和宗旨的情况下可以对这些实施例进行多种变化、修改、替换和变型,本发明的范围由权利要求及其等同物限定。下列实施例中未注明具体条件的实验方法,通常按照常规条件或按照厂商所建议的条件实施检测。
实施例1人源性腺激素型垂体腺瘤干细胞细胞系的构建及鉴定
1、实验材料
Accutase购自于STEMCELL,货号为07920;
红细胞裂解液购自于索莱宝,货号为R1010;
DMEM/F12购自于Gibco,货号为A4192001;
青-链霉素(5000U/mL)购自于Gibco,货号为15070063;
EGF(20ng/mL)购自于Peprotech,货号为400-31-10;
bFGF(20ng/mL)购自于Peprotech,货号为400-42-25;
N2购自于Gibco,货号为A1370701;
B27购自于Gibco,货号为17504044;
免疫荧光用抗体:
一抗:Sox2(abcam,ab97959),1:800稀释
Oct4(abcam,ab19857),1:500稀释
Nestin(abcam,ab105389),1:200稀释
CD133(abcam,ab19898),1:500稀释
FSH(Santa cruz,sc-374452),1:800稀释
二抗:山羊抗兔(abcam,ab150077),1:800稀释
山羊抗小鼠(abcam,ab150115),1:800稀释
2、垂体腺瘤组织来源
本实施例中的垂体腺瘤肿瘤组织来源于1例FSH型垂体腺瘤患者,该患者于2020年11月在天坛医院神经外科肿瘤2病区手术切除病变部位的肿瘤组织。
性腺激素型垂体腺瘤患者的临床信息如下:李某,男,汉族,诊断信息为 FSH型垂体腺瘤。
3、人源性腺激素型垂体腺瘤干细胞细胞系的构建
(1)取材:经蝶或开颅取新鲜的肿瘤标本,放置于含青-链霉素(200U/mL)的 DMEM/F12培养基中。冰上保存,快速运输至细胞间进行培养;
(2)肿瘤单细胞悬液构建:将垂体瘤标本置于无菌培养皿中,使用预冷的PBS 冲洗去除标本表面血块,无菌剪刀去除坏死组织。准备完毕后使用无菌镊及剪刀将肿瘤标本剪碎为1mm3以下的肿瘤碎片。300g离心5分钟,去上清。加入5 倍肿瘤体积的Accutase,37℃水浴20分钟。300g离心5分钟,去上清后加入5 倍肿瘤体积的红细胞裂解液,冰上充分吹打涡旋。重复上述红细胞裂解步骤2遍。加入垂体瘤干细胞培养基,使用40微米无菌筛网去除未消化的大块肿瘤组织;
(3)细胞培养:加入适量的干细胞培养基,所述干细胞培养基为添加20ng/mL EGF、20ng/mL bFGF、1X N2、1X B27、200U/mL青-链霉素的DMEM/F12无血清培养基,每2-3天换液一次,每7天传一代,培养3周以上,即可构建得到稳定的悬浮生长的性腺激素型垂体瘤干细胞球。
4、对构建的性腺激素型垂体瘤干细胞球进行鉴定
对本实施例构建的性腺激素型垂体瘤干细胞球进行免疫荧光染色,以鉴定肿瘤干细胞标志物的表达情况,具体实验步骤如下:
(1)肿瘤细胞贴壁爬片:取适量干细胞球培养悬液,离心浓缩后加入腔室玻片中(Thermo Fisher,154917),每孔约500微升。37℃孵育4小时;
(2)细胞免疫荧光:4%多聚甲醛室温固定20分钟;PBS漂洗5次;0.3%的 PBST为基液的10%山羊血清室温封闭60分钟;10%山羊血清按上述比例稀释干细胞抗体(Sox2、Oct4、Nestin、CD133)与FSH抗体,4℃孵育过夜;PBS漂洗5 次;PBS按适当比例稀释二抗(abcam,ab150077和ab150077,ab150115),室温孵育60分钟;PBS漂洗3次;DAPI室温染色10分钟;组织自发荧光猝灭剂,室温孵育10分钟,PBS漂洗3次;抗猝灭封片。
血清分化后细胞免疫荧光:以干细胞培养基为基础,添加15%FBS胎牛血清的培养基分化培养肿瘤干细胞5天。免疫荧光步骤与上述步骤相同。
5、实验结果
光镜下离体培养18天后的性腺激素型垂体腺瘤干细胞见图1,结果显示,构建得到的性腺激素型垂体腺瘤干细胞具有高体外成球能力;进一步对其进行免疫荧光染色以鉴定肿瘤干细胞标志物的表达情况的结果图见图2,结果显示,构建得到的促卵泡刺激素型垂体腺瘤干细胞高表达肿瘤干细胞标志物Sox2、Oct4、Nestin、CD133(绿荧光),同时红荧光显色的FSH表达较弱。表明构建得到的性腺激素型垂体腺瘤干细胞具有干细胞的特性。
实施例2对人源性腺激素型垂体腺瘤干细胞进行诱导分化
1、实验方法
通过在培养基中添加15%FBS诱导肿瘤干细胞分化,对同期对照培养的肿瘤干细胞及分化细胞拍照计数,同时,对分化的肿瘤细胞进行免疫组化鉴定肿瘤干细胞标志物。
2、实验结果
实验结果显示,添加血清分化后的肿瘤细胞数明显增加(见图3A和B),表明了血清可促进肿瘤干细胞增殖及分化产生肿瘤细胞。对分化的肿瘤细胞进行肿瘤干细胞标志物的鉴定结果显示,分化获得的肿瘤细胞中干性标志物不同程度的降低(绿色荧光为干细胞标志物);部分细胞中FSH的表达量上升(红色荧光为 FSH)(见图4),表明了添加血清后垂体瘤干细胞的干细胞特征降低,向肿瘤细胞方向分化。
以上已对本发明创造的较佳实施例进行了具体说明,但本发明创造并不限于所述实施例,熟悉本领域的技术人员在不违背本发明创造精神的前提下还可做出种种的等同的变型或替换,这些等同的变型或替换均包含在本申请权利要求所限定的范围内。
Claims (8)
1.一种人源性腺激素型垂体腺瘤干细胞细胞系,其特征在于,其保藏在中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC No.21018。
2.如权利要求1所述的人源性腺激素型垂体腺瘤干细胞细胞系的子代细胞。
3.权利要求1所述的细胞系在制备性腺激素型垂体腺瘤细胞模型中的应用。
4.权利要求1所述的细胞系在制备干细胞异种移植的性腺激素型垂体腺瘤动物模型中的应用。
5.权利要求1所述的细胞系在筛选预防和/或治疗性腺激素型垂体腺瘤的药物中的应用。
6.权利要求1所述的细胞系在建立性腺激素型垂体腺瘤发病机制、侵袭机制的研究平台中的应用。
7.权利要求1所述的细胞系在建立耐药机制研究平台中的应用。
8.一种用于治疗性腺激素型垂体腺瘤的候选药物的体外筛选方法,其特征在于,所述方法包括如下步骤:
(1) 将待测物质施用于权利要求1所述的细胞系中;
(2) 若步骤(1)中所述的待测物质施用后,导致体外细胞凋亡或抑制体外细胞增殖,则所述待测物质为治疗性腺激素型垂体腺瘤的候选药物。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110272526.5A CN113025576B (zh) | 2021-03-12 | 2021-03-12 | 一种人源性腺激素型垂体腺瘤干细胞细胞系的构建方法及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110272526.5A CN113025576B (zh) | 2021-03-12 | 2021-03-12 | 一种人源性腺激素型垂体腺瘤干细胞细胞系的构建方法及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113025576A CN113025576A (zh) | 2021-06-25 |
CN113025576B true CN113025576B (zh) | 2022-07-15 |
Family
ID=76468781
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110272526.5A Active CN113025576B (zh) | 2021-03-12 | 2021-03-12 | 一种人源性腺激素型垂体腺瘤干细胞细胞系的构建方法及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113025576B (zh) |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20100173344A1 (en) * | 2006-08-24 | 2010-07-08 | Cedars-Sinai Medical Center | Methods for isolating and using pituitary adenoma stem cells and pituitary adenoma cells |
WO2008067408A2 (en) * | 2006-11-28 | 2008-06-05 | Cedars-Sinai Medical Center | Method of isolating and propagating stem cells from benign tumors |
CN107460170B (zh) * | 2016-06-02 | 2020-11-10 | 任远 | 人垂体腺瘤细胞系的建立及其应用 |
CN106834232A (zh) * | 2016-08-04 | 2017-06-13 | 北京市神经外科研究所 | 人垂体腺瘤细胞系及其用途 |
-
2021
- 2021-03-12 CN CN202110272526.5A patent/CN113025576B/zh active Active
Also Published As
Publication number | Publication date |
---|---|
CN113025576A (zh) | 2021-06-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN113528444B (zh) | 一种用于食管鳞癌上皮细胞的培养基、培养方法及其应用 | |
Hsu et al. | Generation of chordoma cell line JHC7 and the identification of Brachyury as a novel molecular target | |
WO2021088119A1 (zh) | 原代乳腺上皮细胞培养基、培养方法及其应用 | |
CN112080472B (zh) | 一种培养专用于生物医药功能研究的人肺癌类器官3d模型的方法 | |
CN101855339A (zh) | 人类癌症干细胞 | |
US20100173344A1 (en) | Methods for isolating and using pituitary adenoma stem cells and pituitary adenoma cells | |
CN117965431A (zh) | 一种囊泡及其应用 | |
CN109613254B (zh) | 一种用于肿瘤治疗和诊断的靶点标志物pdia2 | |
CN109152799A (zh) | 胰腺干细胞及其用途 | |
CN110496221A (zh) | 抑制dppa3表达的物质在制备预防和治疗癌症的产品中的应用 | |
CN113025576B (zh) | 一种人源性腺激素型垂体腺瘤干细胞细胞系的构建方法及其应用 | |
CN111440800B (zh) | 靶向Galectin-3基因的miRNA-128-3p及其抗胰腺癌的用途 | |
WO2023217128A1 (zh) | 胃粘膜上皮前体样细胞及其制备方法和应用 | |
CN112553289A (zh) | 一种评价car-t细胞有效性的方法 | |
EP4368706A1 (en) | Culture medium and culture method for lung cancer epithelial cells, and application thereof | |
CN107083428B (zh) | Pak5在癌症诊断预后治疗及药物筛选中的应用 | |
CN107460170B (zh) | 人垂体腺瘤细胞系的建立及其应用 | |
CN113633754B (zh) | 卵巢干细胞在制备卵巢抗衰的药物中的用途 | |
CN115466716A (zh) | 一种患者源性口腔黏液表皮样癌类器官的构建方法及应用 | |
CN114752626A (zh) | 一种可逆性永生化ⅱ型肺泡上皮细胞及其构建与应用 | |
US20140128272A1 (en) | Method for Inducing Dormancy of Cancer Tissue-Derived Cell Mass and Method for Evaluating Treating Means with the Use of Cancer-Tissue-Derived Cell Mass | |
CN113201494B (zh) | 一种黏膜黑色素瘤细胞及其用途 | |
CN103267852B (zh) | 成纤维激活蛋白α在制备胰腺癌预后试剂盒中的用途 | |
CN113025577B (zh) | 一种鼠源生长激素型垂体腺瘤干细胞细胞系gh3sc的构建方法及其应用 | |
WO2022016642A1 (zh) | 一种用于肺癌上皮细胞的培养基、培养方法及其应用 |
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 | ||
GR01 | Patent grant | ||
GR01 | Patent grant |