CN102559597B - Adult nephroblastoma HANB cell strain and culturing method and application thereof - Google Patents
Adult nephroblastoma HANB cell strain and culturing method and application thereof Download PDFInfo
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Abstract
The invention discloses an adult nephroblastoma HANB cell strain with the collection number of CCTCC No:C201133. A culturing method comprises the following steps of: culturing in-vitro ascites of a patient suffering from poorly-differentiated anaplastic nephroblastoma; passing; freezing for preserving; and recovering; the cell growing speed is high, the quantity is large, the quality is uniform, and a cell culturing method is simple; as proved by evaluation on the biological characteristics such as cell form, protein structure, chromosome, neoplastic rate and the like of the cell strain, the cell strain keeps the biological characteristics of a primary tumor; and due to the usage of the adult nephroblastoma HANB cell strain to the establishment of a nephroblastoma model, a research model is provided for the occurrence, transfer mechanism and clinical treatment of the nephroblastoma.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of adult nephroblastoma cell strain and cultural method thereof.
Background technology
The nephroblastoma (Wilms ' tumor or Nephroblastoma) claim again Wei Ermusi knurl, 90% betides the children before 6 years old, and its sickness rate accounts for first place in children abdominal tumour, is the common substantive malignant tumour of belly.It originates from original kidney matrix, can form embryonal structure, and adult's morbidity is rarer, and all insensitive to radiotherapy, chemotherapy, grade malignancy is high, easily there is distant metastasis, often prognosis is not good, therefore studies the adult type nephroblastoma and has important clinical meaning.And at present without nephroblastoma immortalized cell line, for the research of its external treatment experimental study, medicaments insensitive Journal of Sex Research and mechanism has brought difficulty.
Therefore, being immortalized, quality homogeneous, biological property retained the nephroblastoma cell strain of the biological characteristics of former tumour be realize clinical treatment correlative study basis and crucial.
Summary of the invention
A technical problem to be solved by this invention is to provide that one has in-vivo tumour Histopathological characteristics, grade malignancy is high, animal tumorigenesis rate is high, the adult nephroblastoma HANB cell strain rapidly of growing.
Another one technical problem to be solved by this invention provides a kind of cultural method for adult nephroblastoma HANB cell strain.
To be solved by this invention also have technical problem to provide a kind of new purposes for adult nephroblastoma HANB cell strain.
Solving the problems of the technologies described above adopted technical scheme is: adult nephroblastoma HANB cell strain, on May 23rd, 2011, be preserved in Chinese Typical Representative culture collection center, and deposit number is CCTCC NO:C201133.
The cultural method step of above-mentioned adult nephroblastoma HANB cell strain is as follows:
1, subculture
By centrifugal ascites in vitro after modification nephroblastoma operation in patients between low differentiation, precipitation is washed with RPMI-1640, obtain primary cell, RPMI-1640 is mixed with by 1640 substratum, 1640 substratum are the commodity of selling on market, by U.S. Life Tech company, produced, sell Dong Ge bio tech ltd, Beijing; On Bechtop, primary cell is placed in to culturing bottle, with containing the RPMI-1640 suspension that massfraction is 5%～20% newborn calf serum, at 37 ℃, CO
2volumetric concentration is to cultivate in the cell culture incubator of 5%, 95% humidity, every 24 hours, change contain massfraction be 5%～15% newborn calf serum RPMI-1640 once, be cultured to Growth of Cells to plateau, complete the cultivation of going down to posterity for the first time of cell, the cell that grows to plateau is gone down to posterity, and its method is as follows:
The trypsinase aqueous solution that is 0.22%～0.28% with massfraction digests, the trypsinase aqueous solution is abandoned in suction, with containing the RPMI-1640 piping and druming bottle wall that massfraction is 1%～5% newborn calf serum, suspension cell, by suspension move in centrifuge tube 800 revs/min centrifugal, obtain cell precipitation, with going down to posterity for the first time, cultural method and condition continue to carry out sub-bottle cultivation in cell culture incubator, treat that Growth of Cells is to 75% of bottle wall area, with the method collecting cell going down to posterity, cell mass is opened with the piping and druming of dimethyl sulfoxide (DMSO) cells frozen storing liquid, obtaining deposit number is the adult nephroblastoma HANB cell strain of CCTCC NO:C201133.
The preparation method of above-mentioned RPMI-1640 is: 10g 1640 substratum, 5.0g HEPES, 3.7g NaHCO
3, 3g L-glutaminate is dissolved in 1L deionized water, after dissolving with 0.22 μ m millipore filter filtration, degerming packing, 4 ℃ of refrigerations.
2, frozen
By the method collecting cell going down to posterity for subculture step 1, cell mass is opened with the piping and druming of dimethyl sulfoxide (DMSO) cells frozen storing liquid, pack in cryopreservation tube and be placed in-20 ℃ of refrigerators frozen 2～6 hours, take out cryopreservation tube, be placed in-80 ℃ of refrigerators frozen 2～12 hours, take out cryopreservation tube, be placed in-120 ℃ of following liquid nitrogen containers and preserve.
Above-mentioned dimethyl sulfoxide (DMSO) cells frozen storing liquid is mixed by the dimethyl sulfoxide (DMSO) of 10mL and the new-born calf serum of 90mL.
3, recovery
Cell in frozen step 2 liquid nitrogen is melted in 40 ℃～42 ℃ water-baths, by cell suspension move in centrifuge tube 800 revs/min centrifugal, cell precipitation moves in Tissue Culture Flask, by go down to posterity cultural method and condition, proceed to cultivate, nutrient solution adopts the RPMI-1640 of 5%～15% newborn calf serum.
In the subculture step 1 of the cultural method of adult nephroblastoma HANB cell strain, on Bechtop, primary cell is placed in to culturing bottle, with the best massfraction that contains be that the RPMI-1640 of 20% newborn calf serum suspends, at 37 ℃, CO
2volumetric concentration is to cultivate in the cell culture incubator of 5%, 95% humidity, every 24 hours, change the best contain massfraction be 10% newborn calf serum RPMI-1640 once, be cultured to Growth of Cells to plateau, complete the cultivation of going down to posterity for the first time of cell, the cell that grows to plateau is gone down to posterity.Going down to posterity in step, the trypsinase aqueous solution that is 0.25% with best in quality mark digests, the trypsinase aqueous solution is abandoned in suction, contain the RPMI-1640 piping and druming bottle wall that massfraction is 5% newborn calf serum by the best, suspension cell, by suspension move in centrifuge tube 800 revs/min centrifugal, obtain cell precipitation, with going down to posterity for the first time, cultural method and condition continue to carry out sub-bottle cultivation in cell culture incubator, treat that cell grows to 75% of bottle wall area, with the method collecting cell going down to posterity, cell mass is opened with the piping and druming of dimethyl sulfoxide (DMSO) cells frozen storing liquid, obtaining deposit number is the adult nephroblastoma HANB cell strain of CCTCC NO:C201133.In recovery step 3, cell in frozen step 2 liquid nitrogen is melted in 40 ℃～42 ℃ water-baths, by cell suspension move in centrifuge tube 800 revs/min centrifugal, cell precipitation moves in Tissue Culture Flask, by go down to posterity cultural method and condition, proceed to cultivate the best RPMI-1640 with 10% newborn calf serum of nutrient solution.
The adult nephroblastoma HANB cell strain that above-mentioned cultural method is cultivated is in the purposes of setting up in nephroblastoma model.Its using method is as follows:
By per generation adult nephroblastoma HANB cell strain centrifugal collecting cell of vitro culture, with the phosphate buffered saline buffer of 0.01mol/L, make cell suspension, adding up to cell count is 10
6, every generation is inoculated respectively nude mice, metering is every 1mL, and Continuous Observation 2 weeks, sets up nephroblastoma animal model, can be used for development, the research of transfer mechanism and the therapeutic evaluation of radiotherapy and chemotherapy medicine clinical treatment of nephroblastoma cell.
Adult nephroblastoma HANB cell strain growth of the present invention is rapid, cultivation is simple, by biological properties such as the cellular form to this cell strain, protein structure, karyomit(e) and tumorigenesis rates, evaluate, confirm that this cell strain has retained the biological characteristics of former tumour substantially, the good model of nephroblastoma basis and clinical study, for generation, transfer mechanism and the clinical treatment of the research nephroblastoma provide research model.
Accompanying drawing explanation
Fig. 1 is that the adult nephroblastoma HANB cell strain that embodiment 1 cultivates amplifies the photo of 40 times under opticmicroscope.
Fig. 2 is that the adult nephroblastoma HANB cell strain that embodiment 1 cultivates amplifies the photo of 200 times under opticmicroscope.
Fig. 3 is the transmission electron microscope photo of the adult nephroblastoma HANB cell strain cultivated of embodiment 1.
Fig. 4 is the growth curve of the adult nephroblastoma HANB cell strain cultivated of embodiment 1.
Fig. 5 is the growth cycle figure of the adult nephroblastoma HANB cell strain cultivated of embodiment 1.
Embodiment
Below in conjunction with drawings and Examples, the present invention is described in more detail, but the invention is not restricted to these embodiment.
In following examples and experiment, the compound method of involved solution is:
RPMI-1640: 10g 1640 substratum, 5.0g HEPES, 3.7g NaHCO
3, 3g L-glutaminate is dissolved in 1L deionized water, after dissolving with 0.22 μ m millipore filter filtration, degerming packing, 4 ℃ of refrigerations.
Contain the RPMI-1640 that massfraction is 1% newborn calf serum: RPMI-1640 is to mix at 9.9: 0.1 with new-born calf serum by volume.
Contain the RPMI-1640 that massfraction is 5% newborn calf serum: RPMI-1640 is to mix at 9.5: 0.5 with new-born calf serum by volume.
Contain the RPMI-1640 that massfraction is 10% newborn calf serum: RPMI-1640 is to mix at 9: 1 with new-born calf serum by volume.
Contain the RPMI-1640 that massfraction is 20% newborn calf serum: RPMI-1640 is to mix at 8: 2 with new-born calf serum by volume.
Digestive ferment: 0.25g trypsinase powder is dissolved in 100ml deionized water, is mixed with massfraction and is 0.25% the trypsinase aqueous solution, filter-sterilized, 4 ℃ of preservations.
0.01M phosphate buffered saline buffer: 8.0g NaCl, 0.2g KCl, 1.56g Na
2hPO
412H
2o, 0.2g KH
2pO
4be dissolved in 1L distilled water, pH value is 7.4,4 ℃ of preservations after autoclaving.
Dimethyl sulfoxide (DMSO) cells frozen storing liquid: 10mL dimethyl sulfoxide (DMSO) is mixed with 90mL new-born calf serum, 4 ℃ of preservations.
Adult nephroblastoma HANB cell strain, deposit number is CCTCC NO:C201133, its cultural method is as follows:
1, subculture
By centrifugal 10 minutes 1000 revs/min of ascites in vitro after modification nephroblastoma operation in patients between low differentiation, discard supernatant liquid, precipitation is washed with RPMI-1640, obtain primary cell, RPMI-1640 is mixed with by 1640 substratum, 1640 substratum are the commodity of selling on market, by U.S. Life Tech company, are produced, and sell Dong Ge bio tech ltd, Beijing; The compound method of RPMI-1640 is: 10g 1640 substratum, 5.0g HEPES, 3.7g NaHCO
3, 3g L-glutaminate is dissolved in 1L deionized water, after dissolving with 0.22 μ m millipore filter filtration, degerming packing, 4 ℃ of refrigerations.On Bechtop, primary cell is placed in to Tissue Culture Flask, with the RPMI-1640 suspension of 20% newborn calf serum, at 37 ℃, CO
2volumetric concentration is 5%, humidity is to cultivate in 95% environment, every 24 hours, renew the RPMI-1640 of 10% fresh newborn calf serum once, be cultured to Growth of Cells to plateau, complete the cultivation of going down to posterity for the first time of cell, the cell that grows to plateau is gone down to posterity, and its method is as follows:
The trypsinase aqueous solution that is 0.25% with massfraction digests, the trypsinase aqueous solution is abandoned in suction, with containing the RPMI-1640 piping and druming bottle wall that massfraction is 3% newborn calf serum, suspension cell, by suspension move in centrifuge tube 800 revs/min centrifugal, obtain cell precipitation, with going down to posterity for the first time, cultural method and condition continuation divide 3 bottles to cultivate in cell culture incubator, treat that Growth of Cells is to 75% of bottle wall area, with the method collecting cell going down to posterity, cell mass is opened with the piping and druming of dimethyl sulfoxide (DMSO) cells frozen storing liquid, obtaining deposit number is the adult nephroblastoma HANB cell strain of CCTCC NO:C201133.
2, frozen
By the method collecting cell going down to posterity for subculture step 1, cell mass is opened with the piping and druming of dimethyl sulfoxide (DMSO) cells frozen storing liquid, pack in cryopreservation tube and be placed in-20 ℃ of refrigerators frozen 2～6 hours, take out cryopreservation tube, be placed in-80 ℃ of refrigerators frozen 2～12 hours, take out cryopreservation tube, be placed in-120 ℃ of following liquid nitrogen containers and preserve.
Above-mentioned dimethyl sulfoxide (DMSO) cells frozen storing liquid is mixed by the dimethyl sulfoxide (DMSO) of 10mL and the new-born calf serum of 90mL;
3, recovery
Cell in frozen step 2 liquid nitrogen is melted in 40 ℃～42 ℃ water-baths, cell suspension move in centrifuge tube 800 revs/min centrifugal, cell precipitation moves in Tissue Culture Flask, by go down to posterity cultural method and condition, proceeds to cultivate, and nutrient solution adopts the RPMI-1640 of 10% newborn calf serum.
Adult nephroblastoma HANB cell strain, deposit number is CCTCC NO:C201133, its cultural method is as follows:
In the subculture step 1 of the cultural method of adult nephroblastoma HANB cell strain, on Bechtop, primary cell is placed in to culturing bottle, with containing massfraction, be that the RPMI-1640 of 5% newborn calf serum suspends, at 37 ℃, CO
2volumetric concentration is to cultivate in the cell culture incubator of 5%, 95% humidity, every 24 hours, change contain massfraction be 5% newborn calf serum RPMI-1640 once, be cultured to Growth of Cells to plateau, complete the cultivation of going down to posterity for the first time of cell, the cell that grows to plateau is gone down to posterity, going down to posterity in step, the trypsinase aqueous solution that is 0.22% with massfraction digests, the trypsinase aqueous solution is abandoned in suction, with containing the RPMI-1640 piping and druming bottle wall that massfraction is 1% newborn calf serum, suspension cell, by suspension move in centrifuge tube 800 revs/min centrifugal, obtain cell precipitation, with going down to posterity for the first time, cultural method and condition continue to carry out sub-bottle cultivation in cell culture incubator, treat that Growth of Cells is to 75% of bottle wall area, with the method collecting cell going down to posterity, cell mass is opened with the piping and druming of dimethyl sulfoxide (DMSO) cells frozen storing liquid, obtaining deposit number is the adult nephroblastoma HANB cell strain of CCTCC NO:C201133, other steps of this step are identical with embodiment 1.In recovery step 3, cell in frozen step 2 liquid nitrogen is melted in 40 ℃～42 ℃ water-baths, cell suspension move in centrifuge tube 800 revs/min centrifugal, cell precipitation moves in Tissue Culture Flask, by go down to posterity cultural method and condition, proceed to cultivate the RPMI-1640 of 5% newborn calf serum for nutrient solution.
Other steps are identical with embodiment 1.
Adult nephroblastoma HANB cell strain, deposit number is CCTCC NO:C201133, its cultural method is as follows:
In the subculture step 1 of the cultural method of adult nephroblastoma HANB cell strain, on Bechtop, primary cell is placed in to culturing bottle, with containing massfraction, be that the RPMI-1640 of 20% newborn calf serum suspends, at 37 ℃, CO
2volumetric concentration is to cultivate in the cell culture incubator of 5%, 95% humidity, every 24 hours, change contain massfraction be 15% newborn calf serum RPMI-1640 once, be cultured to Growth of Cells to plateau, complete the cultivation of going down to posterity for the first time of cell, the cell that grows to plateau is gone down to posterity, going down to posterity in step, the trypsinase aqueous solution that is 0.28% with massfraction digests, the trypsinase aqueous solution is abandoned in suction, with containing the RPMI-1640 piping and druming bottle wall that massfraction is 5% newborn calf serum, suspension cell, by suspension move in centrifuge tube 800 revs/min centrifugal, obtain cell precipitation, with going down to posterity for the first time, cultural method and condition continue to carry out sub-bottle cultivation in cell culture incubator, treat that Growth of Cells is to 75% of bottle wall area, with the method collecting cell going down to posterity, cell mass is opened with the piping and druming of dimethyl sulfoxide (DMSO) cells frozen storing liquid, obtaining deposit number is the adult nephroblastoma HANB cell strain of CCTCC NO:C201133, other steps of this step are identical with embodiment 1.In recovery step 3, cell in frozen step 2 liquid nitrogen is melted in 40 ℃～42 ℃ water-baths, by cell suspension move in centrifuge tube 800 revs/min centrifugal, cell precipitation moves in Tissue Culture Flask, by go down to posterity cultural method and condition, proceed to cultivate the RPMI-1640 of 15% newborn calf serum for nutrient solution.
Other steps are identical with embodiment 1.
The adult nephroblastoma HANB cell strain that above-described embodiment 1～3 cultural method is cultivated is in the purposes of setting up in nephroblastoma model.Its using method is as follows:
By per generation adult nephroblastoma HANB cell strain centrifugal collecting cell of vitro culture, with the phosphate buffered saline buffer of 0.01mol/L, make cell suspension, adding up to cell count is 10
6, every generation is inoculated respectively nude mice, metering is every 1mL, and Continuous Observation 2 weeks, sets up nephroblastoma animal model, can be used for development, the research of transfer mechanism and the therapeutic evaluation of radiotherapy and chemotherapy medicine clinical treatment of nephroblastoma cell.
In order to verify beneficial effect of the present invention, the adult nephroblastoma HANB cell strain that contriver adopts the embodiment of the present invention 1 to cultivate is tested, and various experiment situations are as follows;
1, the form of adult nephroblastoma HANB cell strain
The adult nephroblastoma HANB cell strain of results, with containing the RPMI-1640 suspension that massfraction is 10% newborn calf serum, is made to cell suspension, be inoculated in 6 well culture plates that are placed with autoclaving cover glass, at 37 ℃, CO
2volumetric concentration is to cultivate in the cell culture incubator of 5%, 95% humidity, every 24 hours, change contain massfraction be 10% newborn calf serum RPMI-1640 once, be cultured to Growth of Cells to plateau, nutrient solution is abandoned in suction, the cover glass (being cell climbing sheet) of growing with 0.01M phosphate buffer wash cell 2 times, inhales and abandons damping fluid, adds 95% dehydrated alcohol fixed cell, adopt observation by light microscope cellular form, observations is shown in Fig. 1～2.
The glutaraldehyde phosphoric acid buffer that is first 2.5% with massfraction by the adult nephroblastoma HANB cell strain of results is fixed, the sucrose phosphoric acid buffer that is 7% with massfraction rinses, be fixed on again volume fraction and be in 1% OsO4 (osmic acid), dewater step by step with acetone, epoxy resin embedding, ultrathin section(ing), with lead citrate, acetic acid uranium double staining, adopt JEM1200 type transmission electron microscope observing cellular form, observations is shown in Fig. 3.
From Fig. 1～2, cell is variform, has small circular, fusiformis and Polygons, and indivedual cells have pseudopodium, assembles in the form of sheets growth, and caryoplasm ratio is obvious.As seen from Figure 3, tumour cell is extremely abundant, and nucleus is intensive, core is large, irregular, kernel is many, welt, bag slurry is few, the abundant microvilli of cell surface, visible rrna, rough surfaced endoplasmic reticulum and obvious, abundant fat drip, identical with former knurl body transmission electron microscope observing performance.
2, the growth characteristics of adult nephroblastoma HANB cell strain
Cell is inoculated in to 24 orifice plates by 4 × 10/mL, counts 3 porocyte numbers every day, computation of mean values, continuous 12 days, draws cell growth curve, according to Patterson formula
Td＝T×lg2/lg(Nt/No)
Calculate the cell colony doubling time, in formula, Td is cell doubling time, T be cell by Nt to No required time, Nt cultivates t days time to inoculate the cell count in hole, No is the cell count of inoculating in orifice plate on the same day.After adult nephroblastoma HANB cell strain inoculation, after the 4th day, enter logarithmic phase, within the 9th day, enter plateau, cell doubling time is 45.6 hours.
The cell in vegetative period of taking the logarithm, collect single cell suspension, centrifugal rear Eddy diffusion is used in the phosphate buffered saline buffer of 0.01mol/L in 5mL, 4 ℃ of precoolings, the massfraction that slowly adds again 4 ℃ of precoolings is 95% ethanol 15mL, making final concentration mass percent is 75%, ice bath 30 minutes, with row flow cytometer detection cell DNA content.Flow cytometer records the G1 phase (DNA synthesizes the standby period) cell 83, the G2 phase (mitotic standby period) 9.7, the S phase (synthesis phase of DNA) 7.3, G2/G1 is 1.9, for hypo-triploid cell, cell growth cycle meets the feature of tumour cell.
3, the chromosome analysis of adult nephroblastoma HANB cell strain
Get the nephroblastoma cell of cultivating 48 hours, adding colchicine working fluid to final concentration is 0.02mg/mL, continue to cultivate 4 hours, collecting cell, centrifugal, hypotonic fixing, at vacuum drying oven inner drying, film-making, with the dyeing of Ji's nurse Sa staining, 50 metacinesis phases of microscopy, carry out chromosome analysis.Adult nephroblastoma HANB cell strain went down to posterity through continuous 32 generations, karyomit(e) keeps the chromosomal feature of human tumor: chromosome number is distributed in 53～74, modal number is 62～68, account for 75%, be mainly triploid caryogram, there is most central authorities and submetacentric chromosome, all keep human chromosomal caryogram.
4, with adult nephroblastoma HANB cell strain, set up animal model
In tumor after being inoculated into nude mice test, by after the 3rd, 9 generation adult nephroblastoma HANB cell strain centrifugal collecting cells of vitro culture, with the phosphate buffered saline buffer of 0.01mol/L, make cell suspension, total cell count is 10
6, every generation is inoculated respectively 8 nude mices, metering is 1mL, Continuous Observation 2 weeks, and establish 2 groups, every group 2 of negative controls, the phosphate buffered saline buffer of the 0.01mol/L of injection equal volume.Continuous Observation 2 weeks, test-results is in Table 1.
The tumorigenesis rate of table 1 adult nephroblastoma HANB cell strain
From table 1, with inoculation time, increase knurl body and increase, every composition ratio of outflow is 100% (8/8), and negative control group produces without knurl body.
5, the significant Protein Detection of adult nephroblastoma HANB cell strain
Adopt immunohistochemical markers method (streptavidin-vitamin H amplification system), carry out respectively vimentin and EMA immunohistochemical localization.Tissue slice dewaxing, aquation, it is each 5 minutes that the phosphate buffered saline buffer of 0.01mol/L is washed twice of tissue slice, with the phosphate buffered saline buffer of distilled water or 0.01mol/L, configuring fresh massfraction is 3% hydrogen peroxide, each 5～10 minutes of room temperature closing cell's creep plate and tissue slice, with distillation washing 3 times, antigen retrieval, the phosphate buffered saline buffer of 0.01mol/L is washed 5 minutes, drip normal goats serum confining liquid, room temperature 20 minutes, get rid of unnecessary liquid, drip I anti-, the anti-concentration of I: vimentin 1: 500 (I is anti-to be produced by U.S. DAKO production company), EMA 1: 500, room temperature is spent the night or 37 ℃ 1 hour (4 ℃ spend the night after 37 ℃ of rewarmings 45 minutes) for 1 hour or 4 ℃, the phosphate buffered saline buffer of 0.01mol/L is washed three times, each 2 minutes, drip biotinylation two anti-, 20 ℃～37 ℃ 20 minutes, the phosphate buffered saline buffer of 0.01mol/L wash 3 times each 2 minutes, drip reagent SABC (sale of Wuhan doctor's moral company), 20 ℃～37 ℃ 20 minutes, the phosphate buffered saline buffer of 0.01mol/L wash 4 times each 5 minutes, develop the color with DAB (sale of Wuhan doctor's moral company): DAB colouring reagents box or autogamy chromogenic reagent (under mirror, grasping colour developing degree), distillation washing, Hematorylin is redyed 2 minutes, hydrochloride alcohol differentiation, dehydration, transparent, mounting, microscopy.Experimental result is in Table 2,
The significant protein immunization group result of table 2 adult nephroblastoma HANB cell strain
From table 2, vimentin is all expressed as seen in each generation tumour cell; And EMA this be marked in this tumour all without positive performance, consistent with former tumour immunity group result in body.
Claims (1)
1. an adult nephroblastoma HANB cell strain, its deposit number is CCTCC NO:C201133.
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| CN1336431A (en) * | 2001-08-23 | 2002-02-20 | 北京大学人民医院 | Establishment of immortal human ovary carcinoma cell strain |
| CN1982441A (en) * | 2006-02-16 | 2007-06-20 | 上海市胸科医院 | Human lung adenocarcinoma self-transferring cell strain and its construction |
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| CN1336431A (en) * | 2001-08-23 | 2002-02-20 | 北京大学人民医院 | Establishment of immortal human ovary carcinoma cell strain |
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