CN102424815B - Cell strain from human lung anaplastic cancers and preparation method thereof - Google Patents
Cell strain from human lung anaplastic cancers and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a cell strain from human lung anaplastic cancers. The cell strain is named human lung cancer cell strain 0114, with collection number being CCTCCNO:C201026. The cell strain is prepared from the lung mass excision of the lung cancer patient with pathological type being anaplastic cancer, has typical characteristic of cellular biology of tumor and can carry out continuous passage in vitro for a long time. Meanwhile, the invention also discloses a preparation method of the cell strain.
Description
Technical field
The present invention relates to a kind of human lung carcinoma cell line and preparation method thereof, especially a kind of cell strain that derives from people's lung undifferentiated carcinoma and preparation method thereof.
Background technology
Tumour is one of principal disease of serious harm human health, the research report that the World Health Organization delivers shows, whole world cancer condition will be day by day serious, therefore, the molecular mechanism of further investigation tumorigenesis and transfer, seek more efficiently oncotherapy scheme, become the advanced subject of current medical research.Along with deepening continuously of oncobiology research, the early diagnosis of tumour and prevention level are enhanced, but the height after oncotherapy shifts and high recurrence is still difficult problem that needs to be resolved hurrily in clinical therapy of tumor.In recent years, proposition and the progress thereof of " tumor stem cell " theory go to be familiar with tumour from a brand-new angle.This theory thinks, have the unique stem-like cell subgroup with self and differentiation function of a part in tumour and it be called tumor stem cell, this cell subset just tumour cells of origin and keep the growth of tumour.Therefore, tumor cell line is the important materials of the Biomedical Problems such as research cell carcinogenesis mechanism, metastases, tumor radiotherapy and chemosensitivity molecular basis, sets up and identifies that different tumor cell lines is a job highly significant.
Summary of the invention
The cell strain that the purpose of this invention is to provide a kind of people's of deriving from lung undifferentiated carcinoma, described cell strain have typical cellular biology of tumor characteristic; A kind of preparation method of described cell strain also is provided simultaneously.
For achieving the above object, the technical scheme that the present invention takes is: a kind of cell strain of the people's of deriving from lung undifferentiated carcinoma, called after human lung carcinoma cell line 0114, preserving number are CCTCC NO:C201026.
Described cell strain is observed demonstration under Electronic Speculum, cell surface long wool hair, and iuntercellular fine hair is entwined, and connects closely, and core is large, and the nuclear membrane marginal distribution has heterochromatin, and organoid is more with plastosome and endoplasmic reticulum, and plastosome mostly is zebra line-transect plastochondria.
Described cell strain is observed under inverted microscope two kinds of growthhabits, a kind of is suspension growth, the cell refractivity is better, endochylema is abundant, contains a large amount of refractivity materials, and a kind of is adherent growth, iuntercellular connects closely, the iuntercellular boundary is unclear, closely attaches nondigestible dispersion between cell and plate; Suspension cell can transform to attached cell, in case adherent, just difficult drop-off.
The external doubling time of described cell strain is 42.685h.
The protein expression of described cell strain is characterized as vimentin, CK7 and TTF-1 all expresses the positive.
The chromosome number of described cell strain is the 40-52 bar.
Human lung carcinoma cell line 0114 of the present invention is a strain factor analysis lung undifferentiated cancer cell strain, derives from 59 years old female lung cancer patient's lower-left lung cancer focus.Patient does not accept the treatments such as any chemotherapy, radiotherapy and targeted therapy before undergoing surgery, find in art that lung tumors is positioned at the lower-left lung, and quality is crisp soft, and is a large amount of downright bad, and the profit growth is invaded in the part, and parietal pleura extensively shifts.
The described cell strain that derives from people's lung undifferentiated carcinoma, to excise thing from the lung cancer patient pulmonary masses that histological type is undifferentiated carcinoma to cultivate gained, be accredited as the lung undifferentiated carcinoma through pathology, be the Polygons epithelial cell, have typical cellular biology of tumor characteristic, it can be at external continuous Long Term Passages, external preparation 1 year, pass generation more than 100, cell still growing multiplication is active.
A kind of preparation method who derives from the cell strain of people's lung undifferentiated carcinoma comprises the following steps:
(1) tissue block mechanical dissociation: people's lung undifferentiated carcinoma tissue of excision is dissociated, clean and remove impurity, separate the Parenchyma of Lung Carcinoma tissue, remove normal alveolar and bronchial tissue and stroma, obtain remaining tissue;
(2) collagenase digesting: the remaining tissue that step (1) is obtained is cut into tissue, cleans, and then adds collagenase to tissue, hatches, and filters, and then collects tumour cell, carries out inoculation culture, obtains survivaling cell;
(3) survivaling cell that step (2) is obtained carries out purifying, removes the impurity such as fibrocyte, and the lung carcinoma cell cultured continuously after purifying also goes down to posterity greater than 12 months, namely gets cell strain.
As preparation method's of the present invention preferred implementation, described preparation method comprises the following steps:
(1) tissue block mechanical dissociation: people's lung undifferentiated carcinoma tissue of excision is dissociated, clean impurity such as removing mucus and red corpuscle, separate the Parenchyma of Lung Carcinoma tissue, remove normal alveolar and bronchial tissue and stroma, obtain remaining tissue;
(2) collagenase digesting: the remaining tissue that step (1) is obtained is cut into 1mm
3Tissue, the cleansing tissue sheet allows tissue precipitation, removes supernatant liquor, residue is organized again frittered, then the cleansing tissue fragment adds collagenase to tissue, hatches under 37 ℃ 4-18 hour; Filter, then collect tumour cell, suspension cell in containing the DMEM in high glucose substratum of 20% foetal calf serum carries out inoculation culture, obtains survivaling cell;
(3) survivaling cell that step (2) is obtained carries out purifying, removes the impurity such as fibrocyte, and the cultured continuously and going down to posterity greater than 12 months in containing the DMEM in high glucose substratum of 20% foetal calf serum of the lung carcinoma cell after purifying namely gets cell strain.
As preparation method's of the present invention preferred implementation, described step (1) is: people's lung undifferentiated carcinoma tissue of excision is dissociated into 0.5cm
3The two anti-solution soaking of green grass or young crops-Streptomycin sulphate with 100% move to Biohazard Safety Equipment, with the PBS damping fluid of sterilization repeatedly cleansing tissue remove the impurity such as mucus and red corpuscle, be soaked in again and move under dissecting microscope in substratum, separate the Parenchyma of Lung Carcinoma tissue with two No. 10 syringe needles, remove normal alveolar and bronchial tissue and stroma, obtain remaining tissue.
As preparation method's of the present invention preferred implementation, step (2) is: use aseptic scalper and scissors that the remaining tissue that step (1) obtains is cut into 1mm
3Tissue, with PBS buffer solution for cleaning tissue, allow tissue precipitate, remove supernatant liquor, with aseptic scalper and scissors, residue is organized again and frittered, with PBS cleansing tissue fragment for several times, then add collagenase (50-200 unit/ml, be dissolved in PBS), hatched under 37 ℃ 4-18 hour, by aseptic Stainless Steel Cloth or nylon net filter cell suspension, then collect tumour cell, suspension cell in containing the DMEM in high glucose substratum of 20% foetal calf serum carries out inoculation culture, obtains survivaling cell.More preferably, in step (2), also add the CaCl that 3mM is arranged when adding collagenase
2Add CaCl
2Can improve collagenase to the dissociation efficiency of tissue.
As preparation method's of the present invention preferred implementation, the culture temperature of the lung carcinoma cell in step (3) after purifying is 37 ℃, and atmosphere surrounding is 5%CO
2/ 95% air, humidity are saturated humidity, change a nutrient solution, go down to posterity once in every 3-5 days in every 3 days.
The cell strain that derives from people's lung undifferentiated carcinoma that adopts preparation method of the present invention to obtain, to excise thing from the lung cancer patient pulmonary masses that histological type is undifferentiated carcinoma to prepare gained, be accredited as the lung undifferentiated carcinoma through pathology, be epithelial cell, have typical cellular biology of tumor characteristic, it can be at external continuous Long Term Passages, vitro culture 1 year, pass generation more than 100, cell still growing multiplication is active.Cell strain of the present invention after testing, its general biological characteristics shows, described cell strain is the Polygons epithelioid cell, but the spherical suspension growth of encystation, in case adherent, also can grow, and adherent closely, the contact growth-inhibiting disappears.
Description of drawings
Fig. 1 is cell strain inverted microscope low suspension growth viable cell picture of the present invention.
Fig. 2 is adherent growth viable cell picture under cell strain inverted microscope of the present invention.
Fig. 3 is chromosomal a kind of striograph of cell strain of the present invention.
Fig. 4 is the chromosomal another kind of striograph of cell strain of the present invention.
Fig. 5 is the Electronic Speculum ultrastructure figure of cell strain of the present invention.
Fig. 6 is the aspect graph that cell strain of the present invention is planted in NOD-SCID mouse scapular region subcutaneous transplantation tumor tissue.
Embodiment
For the purpose, technical solutions and advantages of the present invention better are described, the invention will be further described below in conjunction with the drawings and specific embodiments.
Embodiment 1
A kind of cell strain that derives from people's lung undifferentiated carcinoma of the present invention, according to following preparation method's gained:
(1) tissue block mechanical dissociation: the cancerous lung tissue of excision is dissociated to rapidly 0.5cm with the little scissors of sterilizing
3Size, move to Biohazard Safety Equipment with the two anti-solution soaking of 100% green grass or young crops-Streptomycin sulphate, sterilization PBS damping fluid cleansing tissue is repeatedly removed the impurity such as mucus and red corpuscle, be soaked in again and move under dissecting microscope in substratum, with the careful Parenchyma of Lung Carcinoma tissue that separates of two No. 10 syringe needles, remove normal alveolar and bronchial tissue and stroma as far as possible, obtain remaining tissue;
(2) collagenase digesting: remove the stroma in lung cancer specimen under dissecting microscope after, use aseptic scalper and scissors that the remaining tissue of step (1) gained is cut into 1mm
3Tissue is with PBS buffer solution for cleaning tissue.Allow tissue precipitation, remove supernatant liquor, with aseptic scalper and scissors, residue is organized again and frittered, with PBS cleansing tissue fragment several, then add collagenase (50-200 unit/ml is dissolved in PBS), 37 ℃ hatch 4-18 hour after.Can add 3mM CaCl in the latex collagenase
2, to improve collagenase to the dissociation efficiency of tissue.By aseptic Stainless Steel Cloth or nylon net filter cell suspension, to separate cell dispersion, fragment of tissue and larger fragment; Can add fresh collagenase further to dissociate to larger fragment of tissue.After digestion fully, collect tumour cell, suspension cell in containing the DMEM in high glucose substratum of 20% foetal calf serum carries out inoculation culture, obtains survivaling cell;
(3) step (2) gained survivaling cell is carried out purifying, remove the impurity cells such as fibrocyte, the lung carcinoma cell after purifying is cultivated in containing the DMEM in high glucose substratum of 20% foetal calf serum, and culture temperature is 37 ℃, and atmosphere surrounding is 5% CO
2/ 95% air, humidity are saturated humidity, change a nutrient solution in every 3 days, go down to posterity once in every 3-5 days, and cultured continuously also goes down to posterity over 12 months, gets cell strain.
Cancerous lung tissue in described step (1) derives from 59 years old female lung cancer patient's lower-left lung cancer focus.Patient does not accept the treatments such as any chemotherapy, radiotherapy and targeted therapy before undergoing surgery, find in art that lung tumors is positioned at the lower-left lung, and quality is crisp soft, and is a large amount of downright bad, and the profit growth is invaded in the part, and parietal pleura extensively shifts.
The cell strain that obtains in described step (3) is sent to the center preservation of Chinese Typical Representative culture collection on April 1st, 2010, and deposit number is CCTCC NO:C201026, called after human lung carcinoma cell line 0114.
Embodiment 2
The detection of human lung carcinoma cell line 0114 of the present invention is identified
1, the G6PD isozyme detects source of species (by Chinese Typical Representative culture collection Spot detection).
Human lung carcinoma cell line 0114 of the present invention is a strain factor analysis lung undifferentiated cancer cell strain, derives from 59 years old female lung cancer patient's lower-left lung cancer focus.Patient does not accept the treatments such as any chemotherapy, radiotherapy and targeted therapy before undergoing surgery, find in art that lung tumors is positioned at the lower-left lung, and quality is crisp soft, and is a large amount of downright bad, and the profit growth is invaded in the part, and parietal pleura extensively shifts.
2, morphologic observation
The gross morphology of main observation of cell, as general form, caryoplasm ratio, chromatin and kernel size, what etc., and the ordered state of cytoskeletal filament microtubule etc.
2.1 inversion observation by light microscope
Main under the normal growth state gross morphology of observation of cell, as general form, caryoplasm ratio, adhesion characteristics etc.
Be inverted and observe human lung carcinoma cell line 0114 of the present invention under the optics phase microscope, as seen it has two kinds of growthhabits, the part cell is suspension growth, the cell refractivity is better, abundant a large amount of refractivity materials, the part adherent growth of containing of endochylema, iuntercellular connects closely, the iuntercellular boundary is unclear, closely attaches nondigestible dispersion between cell and plate; Suspension cell can transform to attached cell, in case adherent, just be difficult for coming off again.As shown in attached Fig. 1 and 2.
2.2 projection electron microscopic observation
(1) take out sample from 2.5% glutaraldehyde stationary liquid, use 0.2mol/L PBS(pH7.4) rinsing 3 times, each 15min;
(2) 1% OsO
4Rear fixing, room temperature 1h;
(3) 0.2mol/L PBS(pH7.4) rinsing is 3 times, each 15min;
(4) 50%, 70%, 90% and 100% acetone dewaters step by step, every gradient concentration 15min;
(5) soak into: use acetone: resin=1:1 soaks 1h, then uses acetone: resin=1:2 to soak the last virgin resin of 2h and soaks and spend the night;
(6) carry out polymerization after the sample embedding, 70 ℃, 24h;
(7) ultrathin section(ing), thickness 80nm;
(8) after lead-uranium dyeing, upper machine is observed.
The Electronic Speculum ultrastructure shows, human lung carcinoma cell line 0114 cell of the present invention has impressive cell surface long wool hair, and iuntercellular fine hair is entwined, and connects closely.Core is large, a little heterochromatin of nuclear membrane marginal distribution, and organoid is more with plastosome and endoplasmic reticulum, and plastosome mostly is zebra line-transect plastochondria, as shown in Figure 5.
2.3 immunohistochemical methods detects cell marking albumen
Adopt the S-P method to carry out immunohistochemical staining, immunohistochemical staining S-P test kit is available from Fuzhou Maixin biotechnology Development Co., Ltd.Antibody used is: CK7, TTF-1, vimentin, EGFR, VEGF and NSE.
Concrete steps are:
(1) 24 hours in advance creep plates of cell strain to be detected on the sterilization slide glass, after 24 hours, rinse 2 times with the PBS damping fluid, and cold acetone is fixed 15 minutes, is soaked in PBS standby;
(2) get required slide and add under 1 or 50 μ l peroxidase blocking solution (reagent A) room temperatures and hatched 10 minutes, with the activity of blocking-up endogenous peroxydase; PBS rinses three times, each 3 minutes;
(3) remove PBS liquid, every section adds 1 or the normal nonimmune animal serum of 50 μ l (reagent B), hatches under room temperature 10 minutes;
(4) except serum deprivation, every section adds the first antibody of 1 or 50 μ l, hatches under room temperature 60 minutes;
(5) PBS rinses three times, each 3-5 minute; Remove PBS liquid, every section adds one or the 50 biotin labeled second antibody of μ l (reagent C), hatches under room temperature 10 minutes; PBS rinses three times, each 3 minutes;
(6) remove PBS liquid, every section adds 1 or 50 μ l streptomycete antibiotin peroxidase solution (reagent D), hatches under room temperature 10 minutes; PBS rinses three times, each 3 minutes;
(7) remove PBS liquid, every section adds 2 or the 100 freshly prepared DAB of μ l or AEC solution, and microscopically was observed 3-10 minute;
(8) tap water rinses, and Hematorylin is redyed, and PBS or tap water rinse and return indigo plant;
(9) DAB colour developing, section is dry through gradient alcohol dehydration, and dimethylbenzene is transparent, the neutral gum sealing.
Immunohistochemical methods detected result determination methods: press the painted scoring of cell: brown 3 minutes; Brown color 2 minutes; Faint yellow 1 minute; Non-coloring 0 minute.Count positive cell number under same object lens, mark by the quantity of positive cell: a visual field intrinsic color cell 70% be 4 minutes; 51%-75% is 3 minutes; 11%-50% is 2 minutes; 1%-10% is 1 minute; Feminine gender is 0 minute.Two scores multiply each other, and full 3 are divided into "+"; 4 are divided into " ++ "; More than 5 minutes be " +++"."+~ +++" positive expression.
The immunohistochemical staining demonstration of human lung carcinoma cell line 0114 of the present invention, it has similar protein expression to the cell strain that derives from the metastasis tumor tissues, and its outstanding expression characteristic is that vimentin, CK7 and TTF-1 all express the positive.
Vimentin is vimentin, it is a kind of marker of mesenchymal cell wide expression, CK7 and TTF-1 are the markers that the adenocarcinoma of lung of epithelial origin is the most often expressed, the three expresses simultaneously and often shows that tumour has begun to occur the tendency of Epithelial and stromal sample conversion (EMT), the transfer ability of tumour cell improves, this points out from the side, and human lung carcinoma cell line 0114 is that a strain has the cell strain than high metastatic potential.
3, growth and proliferation of cell
Detect cell growth curve, cell division index, doubling time, cell cycle time.
3.1 mtt assay is measured growth and proliferation of cell and to the susceptibility of medicine
(1) get 96 porocytes and prepare plate, add 0.1ml in every hole and contain 2 * 10
4～10 * 10
4The preparation liquid of target cell (DMEM that contains 10% calf serum prepares liquid), preparation is 2-3 hour in the saturation vapour carbonic acid gas of 37 ℃ of 5% CO2 prepares case, allows cell attachment;
(2) prepare 0.1～100 times of successive configuration medicine of liquid with DMEM, every hole adds medicine and the cell to be checked of 0.1ml dilution, 3 repeating holes of each extent of dilution.9 of control wells: 3 positive control holes, every hole add the DMEM that 0.1ml contains 1000 times of medicines and prepare liquid and cell, 3 negative control holes, every hole adds the 0.1ml DMEM that does not contain medicine and prepares liquid and cell, 3 blank holes, every hole adds 0.1ml DMEM and prepares liquid, does not add cell.At 37 ℃ of 5% CO
2The saturation vapour carbonic acid gas prepare and prepared in case 24～48 hours or predetermined time;
(3) suck preparation liquid, with PBS washing once (as being suspension cell, should before sucking supernatant liquor centrifugal preparation plate);
(4) every hole adds 0.1ml PBS and 10 μ l MTT dye liquors, at 37 ℃ of 5% CO
2The saturation vapour carbonic acid gas prepare and prepare 4～6 hours in case;
(5) every hole adds 0.1ml acidifying Virahol, and also the available 10mmol/L HCl that contains 10% SDS replaces the acidifying Virahol, and the mixing that vibrates on vibrator allows reduzate fully dissolve.Put on enzyme connection detector and measure optical density(OD) (OD) value, detect wavelength 570nm, reference wavelength 630nm.To the mapping of diluted sample degree, standard of comparison curve and testing sample curve can be tried to achieve the content of cytokine in testing sample with the OD value.
3.2 the cell flow cytometer detects cell cycle and apoptosis
Flow cytometer is that (Beckman-Ku Erte) company produces U.S. BECKMAN-COULTER.Machine models: ELITE, optical maser wavelength 488nm, power 15 MW.Get 10
6Fixing cell washes twice with PBS, goes to add after supernatant 300 μ l DNA dye liquors (to include propidium iodide 100 μ g/ml and RNase 20 units/ml), room temperature 30min.Upper machine: adjust instrument with fluorescent microsphere (U.S. BECKMAN-COULTER company) after laser tube preheating 30min, make the HCV value of the signal that each amplifier receives＜2%, collect 12000 cells, the DNA cycle analysis calculates the percentage ratio of each phase of cell at last with the MULTYCYCLE software of U.S. PHEONIX company.In addition, apoptosis is to carry software processes with instrument, directly draws the apoptosis rate of cell.
The external doubling time of human lung carcinoma cell line 0114 of the present invention is 42.685h.
4, nucleus type analysis
Detect the caryogram characteristics, the having or not of karyomit(e) quantity, marker chromosomes, banding pattern etc.
4.1 prepare: colchicine (colchicine), physiological saline are mixed with 10 μ g/ml concentration, 15 pounds of sterilizations in 20 minutes, and packing is put-20 ℃; Hypotonic medium: 0.35%KCl; Stationary liquid (Carnoy stationary liquid) methyl alcohol: glacial acetic acid (3:l), interim preparation; Giemsa working fluid: 1 part of stoste and 10 parts of phosphoric acid buffers, interim preparation.
4.2 colchicine is processed: stopped preparation front 2-4 hour, and added colchicine (drip 2 with No. 5 needle points of 1ml syringe, making final concentration is 0.07 μ g/ml) in preparation liquid.
4.3 karyomit(e) preparation
(1) collecting cell: prepared product is all changed in clean centrifuge tube, with the centrifugal 8-10 of 1000rpm minute, abandon supernatant liquor;
(2) hypotonic processing: add the hypotonic medium 8ml of 37 ℃ of pre-temperature in the graduated centrifuge tube, use the dropper mixing, put in 37 ℃ of waters bath with thermostatic control hypotonic 15-25 minute;
(3) pre-fix: add the 0.5ml stationary liquid after hypotonic, gently centrifugal 8-10 minutes of 1000rpm after mixing;
(4) one is fixing: abandon supernatant liquor, add the 5ml stationary liquid, mixing gently, standing 20 minutes; 1000rpm is centrifugal, abandons supernatant liquor;
(5) two fixing, three fixing: same fixing;
(6) suspension processed: after abandoning supernatant liquor, visual cell's quantity adds appropriate stationary liquid to make cell suspension;
(7) drip sheet: draw cell suspension on the high slide glass that drops in a dried and clean of 10-20cm, featheriness is loose, and gas is done;
(8) dyeing: 1:10 Giemsa dyeing 5-10 minute, unnecessary dye liquor is removed in thin washing, and gas is done;
(9) microscopy: seek good dispersion, the moderate division phase of dyeing, oily Microscopic observation chromosome morphology and counting under low power lens.
The chromosome number of human lung carcinoma cell line 0114 of the present invention is few than other lung cancer cell line, and to the record analysis that the cells of 50 division stages carries out, chromosome number is from 40-52, and the karyomit(e) median is 46, mostly is hypodiploid.Do not find marker chromosomes, as shown in accompanying drawing 3 and 4.
5, allosome animal inoculation pvaccination
Inoculating cell suspension in the allosome animal body is observed into the knurl ability.Adopt hypodermic method, pinched gently skin with left hand thumb and forefinger during injection, the right hand is held syringe syringe needle is thrust, and injects at mouse back or forelimb oxter after fixing.
10
6Individual human lung carcinoma cell line of the present invention 0114 cell seeding is subcutaneous at the NOD-SCID mouse scapular region in 54 ages in week, the transplanted tumor of diameter 2mm all appears in the 3rd week plantation position, after 2 months, the knurl body increases to 1.2cm, puts to death animal after anesthesia, the transplanted tumor tissue dyes through embedded section HE, form as shown in Figure 6, cell volume is huge, core is large, kernel is obvious, and is similar to the tissue of patient in source.
Last institute should be noted that; above embodiment is only in order to illustrate technical scheme of the present invention but not limiting the scope of the invention; although with reference to preferred embodiment, the present invention has been done detailed description; those of ordinary skill in the art is to be understood that; can modify or be equal to replacement technical scheme of the present invention, and not break away from essence and the scope of technical solution of the present invention.
Claims (6)
1. a cell strain that derives from people's lung undifferentiated carcinoma, is characterized in that, called after human lung carcinoma cell line 0114, preserving number are CCTCC NO:C201026.
2. the cell strain that derives from people's lung undifferentiated carcinoma as claimed in claim 1, it is characterized in that, described cell strain is observed demonstration under Electronic Speculum, cell surface long wool hair, iuntercellular fine hair is entwined, and connects closely, core is large, the nuclear membrane marginal distribution has heterochromatin, and organoid is more with plastosome and endoplasmic reticulum, and plastosome mostly is zebra line-transect plastochondria.
3. the cell strain that derives from people's lung undifferentiated carcinoma as claimed in claim 1, it is characterized in that, described cell strain is observed under inverted microscope two kinds of growthhabits, and a kind of is suspension growth, the cell refractivity is better, endochylema is abundant, contains a large amount of refractivity materials, and a kind of is adherent growth, iuntercellular connects closely, the iuntercellular boundary is unclear, closely attaches nondigestible dispersion between cell and plate; Suspension cell can transform to attached cell, in case adherent, just difficult drop-off.
4. the cell strain that derives from people's lung undifferentiated carcinoma as claimed in claim 1, is characterized in that, the external doubling time of described cell strain is 42.685h.
5. the cell strain that derives from people's lung undifferentiated carcinoma as claimed in claim 1, is characterized in that, the protein expression of described cell strain is characterized as vimentin, CK7 and TTF-1 all expresses the positive.
6. the cell strain that derives from people's lung undifferentiated carcinoma as claimed in claim 1, is characterized in that, the chromosome number of described cell strain is the 40-52 bar.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1950285A1 (en) * | 2005-10-05 | 2008-07-30 | GenomIdea Inc. | Isolated human cell, method for obtaining the same and their use |
| CN102250840A (en) * | 2010-05-18 | 2011-11-23 | 上海睿智化学研究有限公司 | Human pancreatic cancer cell line and its application |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1950285A1 (en) * | 2005-10-05 | 2008-07-30 | GenomIdea Inc. | Isolated human cell, method for obtaining the same and their use |
| CN102250840A (en) * | 2010-05-18 | 2011-11-23 | 上海睿智化学研究有限公司 | Human pancreatic cancer cell line and its application |
Non-Patent Citations (2)
| Title |
|---|
| 宋春林 等.肺癌细胞株肺癌相关基因差异表达分析.《江西医学检验》.2004,第22卷(第5期), |
| 肺癌细胞株肺癌相关基因差异表达分析;宋春林 等;《江西医学检验》;20041231;第22卷(第5期);392-394,457 * |
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