CN109593720A - A kind of immortal human squamous nipple type craniopharyngioma cell strain and its application - Google Patents
A kind of immortal human squamous nipple type craniopharyngioma cell strain and its application Download PDFInfo
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Abstract
The invention discloses a kind of craniopharyngioma cell strains, or the derived cell strain of the craniopharyngioma cell strain, it is characterized by: the deposit number of the craniopharyngioma cell strain is CGMCC 16214, the craniopharyngioma cell strain, which is behaved, immortalizes squamous nipple type craniopharyngioma cell strain, cell state is good, it can infinitely pass on, no change has taken place for its benign organisms characteristic simultaneously, a good research platform is provided for the basic research of craniopharyngioma, to craniopharyngioma Biological characteristics, the occurrence and development of tumour, Etiologic Mechanism, clinical treatment measure etc. is significant.
Description
Technical field
The present invention relates to field of biotechnology, and in particular to a kind of immortal human squamous nipple type craniopharyngioma cell strain and
It is applied.
Background technique
Craniopharyngioma is one of most common tumour in encephalic saddle area, is divided into glaze epitheliated type craniopharyngioma and squamous nipple type
Craniopharyngioma.WHO is set to I grades of tumours of WHO, i.e. benign tumour.However, this benign tumour, divides because it is located in human body
Secrete maincenter: hypothalamic pituitary axis, therefore, operating difficulty are very big, postoperative easy recurrence.Tumor by local invasive growth feature is
The main reason for postoperative easy recurrence, invasion mechanism are also the emphasis of lot of domestic and foreign scholar research.But tumor by local is invaded
And recurrence molecular mechanism it is not yet completely clear so far, an important reason be squamous nipple type craniopharyngioma belong to it is benign
Tumour, stabilization can be passed on permanently so that the cell strain of scientific research is deficient.
Humanized's craniopharyngioma cell line is research craniopharyngioma Biological characteristics, the occurrence and development of tumour, etiology and pathology
One of means of crucial importance such as mechanism, clinical treatment exploration and assessment.Craniopharyngioma belongs to the benign tumour of epithelial cell origin,
In vitro culture establishes that stable cell line is especially hard, and the further investigation for resulting in craniopharyngioma cytology level indirectly is very few,
To the translational medicine research that basis is combined with clinic is carried out in a deep going way, there are serious puzzlements.Early in 1997, Germany
Professor Honegger reports for the first time successfully turns out enamel type craniopharyngioma cell, demonstrates progestational hormone in short-term cell culture
Can be in conjunction with the progesterone receptor of tumor cell surface, and tumour is promoted to rise in value；Then in 2005, German Ulfarsson
Professor turns out enamel type craniopharyngioma cell using trypsin digestion, and confirms that insulin-like growth factor I (IGF-1) can promote
Into the proliferation of its tumour cell；German professor Holsken is using the kerationcyte culture medium culture enamel type cranium optimized within 2010
Pharyngeal canal cell, and confirm that being overexpressed CTNNBl gene can promote the migration of tumour.At home, the The 2nd Army Medical College Long March in 2010
Hospital professor Jiang Rongcai confirms bleomycin in body for the first time using mixing pancreatin and collagenase digesting culture craniopharyngioma cell
The outer proliferation for inhibiting tumour cell；Huaxi Hospital Attached to Sichuan Univ Xu Jianguo professors in 2011 and professor Luo Lin report turn out respectively
Craniopharyngioma primary cell, the former confirms that growth hormone promotes the proliferation of tumour, and tamoxifen then inhibits tumour growth, the latter
Confirm that vitamin A acid can induce primary craniopharyngioma Apoptosis.2013, Hospital of Southern Medical University Qi Songtao professor after
Continuous improvement cell primary cultural method, gropes the culture medium for being more suitable for craniopharyhgeal canal tumor cell growth, in vitro culture can pass for 6~9 generations.
The same year, West China Hospital professor You Chao use for reference professor Holsken and constantly improve craniopharyhgeal canal using the spongiocyte culture medium of commercialization
The culture of oncocyte.2015, Navy General Hospital professor Tian Zengmin report digested differential attachment method using pancreatin, successfully turned out
Craniopharyngioma cell.The above are the craniopharyngioma cell culture processes of current domestic and foreign literature report, carefully go into seriously, and discovery exists
Problems: most of craniopharyngioma cell culture reported in the literature is limited only to 1st generation；Some craniopharyngiomas reported in the literature
There are the pollutions of more fibroblast for cell；The actually non-craniopharyngioma cell of the cell that some document reports are cultivated, and
It is tumour associated fibroblast cell；In addition, the craniopharyngioma cellular morphology of each document report culture is different, i.e. the morphology of cell
Corresponding variation has occurred with stability.
Optimize the cell primary culture of craniopharyngioma, especially activity of cell biology, form and stablizes proliferation to pass
It is important；Establishing stable, cloning craniopharyngioma cell strain, oneself becomes urgent need, provides a kind of stable immortalization craniopharyngioma
Cell line is significant to the occurrence and development of craniopharyngioma Biological characteristics, tumour, Etiologic Mechanism, clinical treatment measure etc..
Summary of the invention
The purpose of the present invention is to provide a kind of immortal human squamous nipple type craniopharyngioma cell strain and its applications.
The technical solution used in the present invention is:
The derived cell strain of a kind of craniopharyngioma cell strain or the craniopharyngioma cell strain, the craniopharyngioma cell
The deposit number of strain is CGMCC 16214, and the preservation time is on August 2nd, 2018, and depositary institution is Chinese microorganism strain preservation
Administration committee's common micro-organisms center.
Further, the craniopharyngioma cell strain is immortal human squamous nipple type craniopharyngioma cell strain.
Further, the immortal human squamous nipple type craniopharyngioma cell strain and its patient are same Species origin,
There is BRAF V600E mutation in primary cell, immortalized cells and the tumor tissues in source.
Further, the immortal human squamous nipple type craniopharyngioma cell strain in vitro can be with continuous passage, mesh generation
Number is more than 50 generations, and external lasting culture was more than 9 months, and in succeeding generations, cellular morphology, growth kinetics and hereditary feature are steady
Fixed, by liquid nitrogen cryopreservation, the cell shape after recovery is remained unchanged, and is one plant of stable immortalized cell line.
Further, the cell population doublings time of the immortal human squamous nipple type craniopharyngioma cell strain is 48 small
When, cell passage is carried out after culture 3 days, multiplication rate is stablized.
Further, the immortal human squamous nipple type craniopharyngioma cell strain is seeded to severe Immune deficient mice cranium
Interior, tumorigenesis rate is 100%.
Above-mentioned craniopharyngioma cell strain is in the application established in craniopharyngioma animal model.
Above-mentioned craniopharyngioma cell strain is in the application established in craniopharyngioma mechanism research platform.
Above-mentioned craniopharyngioma cell strain is in the application established in craniopharyngioma medicament research and development and Screening Platform.
Above-mentioned craniopharyngioma cell strain is in the application established in craniopharyngioma immunology and microbiological research platform.
The beneficial effects of the present invention are:
A kind of people is disclosed in the present invention and immortalizes squamous nipple type craniopharyngioma cell strain, and cell state is good, can be with
Unlimited passage, while no change has taken place for its benign organisms characteristic, provides one very well for the basic research of craniopharyngioma
Research platform, to meanings weights such as the occurrence and development of craniopharyngioma Biological characteristics, tumour, Etiologic Mechanism, clinical treatment measures
Greatly.
Detailed description of the invention
Fig. 1 is primary cell Pan-CK Fluorescence Identification result figure；
Fig. 2 is the primary cell after secondary culture；
Fig. 3 is that PCR verifies primary cell VS immortalized cells SV40mRNA expression difference results；
Fig. 4 is that WB verifies primary cell VS immortalized cells SV40 protein expression level difference results；
Fig. 5 is that immortalized cells are passed on more than the Fluorescence Identification (Pan-CK) after 30 generations；
Fig. 6 is Braf V600E abrupt climatic change result；
Fig. 7 is primary cell VS immortalized cells cell cycle, cell Proliferation curve and cell ageing result；
Fig. 8 is immortalized cells nude mice encephalic tumor formation result.
Specific embodiment
It is thin unexpectedly to obtain one plant of people's immortalization squamous nipple type craniopharyngioma by extensive and in-depth research by inventor
Born of the same parents' strain, deposit number are CGMCC 16214, and the preservation time is on August 2nd, 2018, and depositary institution is Chinese microorganism strain guarantor
Hide administration committee's common micro-organisms center.
Below in conjunction with embodiment, the present invention is further explained, it should be appreciated that following embodiment be merely to illustrate the present invention and
It is not used in and limits the scope of the invention.
1 people of embodiment immortalizes the separation of squamous nipple type craniopharyngioma cell strain and builds strain
1. the squamous nipple type craniopharyngioma cut off in pair operation is drawn materials, aseptically, tumor tissues are put
Enter and saved in dry ice chest, is quickly sent to laboratory and is cultivated.
2. cell primary culture
(1) cell culture instrument and super-clean bench Conventional ultraviolet irradiate 30min, and the scaly epithelium type cranium of operation excision is swallowed
Tuberculation tissue removes arachnoid, endocranium, coagulation charred tissue, with the PBS containing 100U/mL penicillin, 100U/mL streptomysin
Washing 3 times, each 3min；
(2) tumor tissues after washing are put into penicillin, are sufficiently shredded with eye scissors, size about 1mm3；
(3) appropriate 0.25% (containing EDTA) pancreatin is added in penicillin bottle, digests tissue block 30min in 37 DEG C of incubators；
(4) 70 μm of postdigestive tissue mass suspensions of strainer filtering；
(5) appropriate Australia fetal calf serum (FBS) is added and terminates pancreatin digestion；
(6) the sticky cell filtrate after appropriate PBS dilute filtration is added；
(7) after being centrifuged (1200 turns/min) 3min, supernatant is abandoned；
(8) cell precipitation is resuspended in complete medium (containing 10%FBS), and plantation is cultivated in culture bottle, 37 DEG C of incubators；
It is more than 90% to cell density after (9) 3~5 days, carries out cell passage.
3. the identification of primary cell
(1) when cell is passed on every time, using differential attachment method, (fibroblast, vascular endothelial cell are completely adherent
Time < 2 hours, tumour cell adherent time > 4 hours completely) carry out purified tumor cell；Tumour is epithelial cell, Pan-
CK specificity epithelium marker can distinguish tumour cell and Stromal fibroblasts, immunocyte, vascular endothelial cell very well
Deng；
(2) it chooses the primary cell of the 3rd generation after purification and carries out identified by immunofluorescence；
(3) culture medium is sopped up, PBS is rinsed 2~3 times；
(4) 4% paraformaldehydes fix cell 10min；
(5) paraformaldehyde is sopped up, PBS is rinsed 2~3 times；
(6) 0.05%Triton rupture of membranes 10min is added；
(7) Triton is sopped up, PBS is rinsed 2~3 times；
(8) 10% lowlenthal serum room temperature is added to close 1 hour；
(9) lowlenthal serum is sopped up, primary antibody Pan-CK (Abcam ab7753 1:200) after dilution is added；
(10) 4 DEG C of refrigerator overnights are incubated for antibody；
(11) wet box is taken out, room temperature 10min is restored, PBS is rinsed 2~3 times；
(12) PBS is sopped up, fluorescence secondary antibody (Life 1:1000) after dilution is added in light protected environment；
(13) room temperature is protected from light incubation 1 hour (following operation carries out all in light protected environment)；
(14) fluorescence secondary antibody is sopped up, PBS is rinsed 2~3 times；
(15) configured DAPI (Roche) is added, is incubated at room temperature 5min；
(16) DAPI is sopped up, PBS is rinsed 2~3 times；
(17) it is added dropwise and prevents being quenched mountant (DACO) in right amount, carry out mounting, be kept in dark place；
(18) it takes pictures under fluorescence microscope, cell Pan-CK Fluorescence Identification result figure is as shown in Figure 1.
4. immortalized cell line is established
(1) it plants after primary cell passage in 96 orifice plates；
(2) it when cell density about 60% (cell enters Exponential growth stage), carries out slow virus carrier and transfects cell；
(3) for the cell after secondary culture 1 day and 3 days as shown in Fig. 2, after 3 days, cell density has covered with 96 orifice plates；
(4) according to limiting dilution assay, single cell clone culture is carried out after each 96 orifice plate passage, is screened after large-scale culture
People's squamous nipple type craniopharyngioma immortalized cell line in good condition and stable；
(5) it carries out changing liquid every 3 days 96 orifice plates, after persistently cultivating 20~30 days, selects single cell clone success and cell
Form has the characteristics that the cell of epithelial cell carries out amplification passage；
(6) the cell passage number after transfecting is more than (remarks: primary craniopharyngioma cell passage algebra < 10 after 10 generations
Generation, usually 3~7 generation, and cell starts the morphologic change such as vacuole, expansion occur after 5 generations), before comparing virus transfection
2 kinds of craniopharyngioma cells afterwards carry out WB and PCR verifying primary cell and immortalized cells SV40 differential expression (result such as figure
3, shown in 4), while the cell colony of high expression SV40 gene is selected, carrying out cell Pan-CK identified by immunofluorescence, (result is as schemed
Shown in 5)；
(7) select that cell state is good, increment is stablized, the cell colony of albumen and all high expression SV40 of gene, continue into
Row cell culture, passage；
(8) cell is persistently passed on more than 30 more than generation, be finally considered as people's squamous nipple type craniopharyngioma immortalized cells at
Function.
The detection of 2 immortal human squamous nipple type craniopharyngioma cell strain biological characteristics of embodiment
(1) abrupt climatic change: pathological diagnosis is squamous nipple type craniopharyngioma, takes the fresh cryopreserved tissue in part, corresponding patient
Primary cell, the cell after immortalizing successfully, mouse encephalic tumor formation success after tumor tissues carry out BRAF (V600E) mutation survey
Sequence (generation sequencing approach), as a result as shown in fig. 6, showing that above 4 samples have mutation.
(2) cell proliferation rate immortalized cells VS primary cell: is detected by cell streaming period and CCK-8 method
(result is as shown in Fig. 7 A, B, E), immortalized cells cell proliferation speed is faster than primary cell as the result is shown；The inspection of β glucosides enzyme dyeing
It surveys senile cell (result is as shown in Fig. 7 C, D), senile cell is significantly more than the tumour after immortalizing in primary cell as the result is shown
Cell.
(3) people's squamous nipple type craniopharyngioma immortalized cells encephalic original position tumor formation: after cell tryptase enzymic digestion, centrifugation is gone
Clearly, a small amount of PBS is added, cell is resuspended, 6 μ L of micro syringe suction of cells suspension (total about 10 cells) is fixed by solid
Into instrument injection joint severe Immune deficient mice (NOD/SCID) brain parenchym, after continuing raising 1.5 months, mouse cranium brain is carried out
Whether MRI scan, observation encephalic tumor formation succeed, this seminar carries out 10 Immune deficient mices in total, tumor formation rate 100% (6/6),
Mouse is put to death under narcosis, takes and fixes 48 hours with 10% neutral formalin after mouse brain, is carried out after automatic dehydrator dehydration
Specimens paraffin embedding slices dye (result is as shown in Figure 8), prompt tumor formation success in situ.
Claims (10)
1. the derived cell strain of a kind of craniopharyngioma cell strain or the craniopharyngioma cell strain, it is characterised in that: the cranium pharynx
The deposit number of tuberculation cell strain is CGMCC 16214.
2. craniopharyngioma cell strain according to claim 1, it is characterised in that: the craniopharyngioma cell strain is to immortalize
People's squamous nipple type craniopharyngioma cell strain.
3. craniopharyngioma cell strain according to claim 2, it is characterised in that: the immortal human squamous nipple type cranium pharynx
Tuberculation cell strain and its patient are same Species origin, and primary cell, immortalized cells and the tumor tissues in source exist
BRAF V600E mutation.
4. craniopharyngioma cell strain according to claim 2, it is characterised in that: the immortal human squamous nipple type craniopharyhgeal canal
Tumor cell strain in vitro can be with continuous passage, and mesh algebra is more than 50 generations, and external lasting culture was more than 9 months, in succeeding generations,
Cellular morphology, growth kinetics and hereditary feature are stablized, and by liquid nitrogen cryopreservation, the cell shape after recovery is remained unchanged, and are one
The stable immortalized cell line of strain.
5. craniopharyngioma cell strain according to claim 2, it is characterised in that: the immortal human squamous nipple type craniopharyhgeal canal
The cell population doublings time of tumor cell strain is 48 hours, and cell passage is carried out after culture 3 days, and multiplication rate is stablized.
6. craniopharyngioma cell strain according to claim 2, it is characterised in that: the immortal human squamous nipple type cranium pharynx
Tuberculation cell strain is seeded to severe Immune deficient mice encephalic, and tumorigenesis rate is 100%.
7. the described in any item craniopharyngioma cell strains of claim 1~6 are in the application established in craniopharyngioma animal model.
8. the described in any item craniopharyngioma cell strains of claim 1~6 are in establishing craniopharyngioma mechanism research platform
Application.
9. the described in any item craniopharyngioma cell strains of claim 1~6 are establishing craniopharyngioma medicament research and development and Screening Platform
In application.
10. the described in any item craniopharyngioma cell strains of claim 1~6 are establishing craniopharyngioma immunology and microbiology is ground
Study carefully the application in platform.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115820537A (en) * | 2022-11-28 | 2023-03-21 | 创芯国际生物科技(广州)有限公司 | Craniopharyngioma organoid, culture medium and culture method |
| CN117637185A (en) * | 2024-01-25 | 2024-03-01 | 首都医科大学宣武医院 | Image-based craniopharyngeal tube tumor treatment auxiliary decision-making method, system and equipment |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100003265A1 (en) * | 2006-09-11 | 2010-01-07 | University Of Florida Research Foundation | Isolation, expansion and uses of tumor stem cells |
| WO2013011513A1 (en) * | 2011-07-21 | 2013-01-24 | Ramot at Tel- Aviv University Ltd. | Methods of determinig enzyme activity in preserved cell samples |
| CN104956226A (en) * | 2013-01-25 | 2015-09-30 | 艾克斯赛尔生物科学公司 | Methods, compositions, kits, and systems for selective enrichment of target cells |
-
2018
- 2018-11-20 CN CN201811386199.0A patent/CN109593720A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100003265A1 (en) * | 2006-09-11 | 2010-01-07 | University Of Florida Research Foundation | Isolation, expansion and uses of tumor stem cells |
| WO2013011513A1 (en) * | 2011-07-21 | 2013-01-24 | Ramot at Tel- Aviv University Ltd. | Methods of determinig enzyme activity in preserved cell samples |
| CN104956226A (en) * | 2013-01-25 | 2015-09-30 | 艾克斯赛尔生物科学公司 | Methods, compositions, kits, and systems for selective enrichment of target cells |
Non-Patent Citations (3)
| Title |
|---|
| HAO LIU ET AL.: "Establishment of primary cultures of craniopharyngioma cells", 《NEURAL REGENERATION RESEARCH》 * |
| YI LIU ET AL.: "Establishing a papillary craniopharyngioma cell line by SV40LT‑mediated immortalization", 《PITUITARY》 * |
| 汪潮湖: "颅咽管瘤原代细胞培养改良及永生化细胞系建立", 《中国优秀博硕士学位论文全文数据库（硕士） 医药卫生科技辑》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115820537A (en) * | 2022-11-28 | 2023-03-21 | 创芯国际生物科技(广州)有限公司 | Craniopharyngioma organoid, culture medium and culture method |
| CN115820537B (en) * | 2022-11-28 | 2023-12-12 | 创芯国际生物科技(广州)有限公司 | Craniopharyngeal pipe tumor organoid, culture medium and culture method |
| CN117637185A (en) * | 2024-01-25 | 2024-03-01 | 首都医科大学宣武医院 | Image-based craniopharyngeal tube tumor treatment auxiliary decision-making method, system and equipment |
| CN117637185B (en) * | 2024-01-25 | 2024-04-23 | 首都医科大学宣武医院 | Image-based craniopharyngeal tube tumor treatment auxiliary decision-making method, system and equipment |
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