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1.
Xavier Léry Joseph Giannotti Ahmed Taha Marc Ravalec Said Abol-Ela 《In vitro cellular & developmental biology. Animal》1997,33(8):640-646
Summary A newly established cell line was obtained from the culture of embryonic cells of the potato tuber moth Phthorimaea operculella in low temperature conditions (19° C) using modified Grace’s medium supplemented with 10% fetal bovine serum. The population
doubling time was about 80 h when cells were cultivated at 19°C and 38 h at 27° C. The cell line had a relatively homogeneous
population consisting of various sized spherical cells. The cells were cultivated for more than 25 passages. Their polypeptidic
profile was different from profiles of other P. operculella cell lines we previously described and from other lepidopteran cells. The new cell line was designated ORS-Pop-95.
The complete replication of the potato tuber moth granulosis virus (PTM GV) was obtained in vitro by both viral infection and DNA transfection. PTM GV multiplied at a significant level during several passages of the cell
line that was maintained at 19° C. As long as the cells were maintained at 19° C, virus multiplication could also be obtained
at the same rate at 27° C. To compare PTM GV multiplied both in vivo and in vitro, we used morphological identification, serological, DNA probe diagnosis and endonuclease digest profile analysis and confirmed
the identity of the virus. 相似文献
2.
Establishment of a cell line with high transfection efficiency from zebrafish Danio rerio embryos and its susceptibility to fish viruses
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Y. L. Jin L. M. Chen Y. Le Y. L. Li Y. H. Hong K. T. Jia M. S. Yi 《Journal of fish biology》2017,91(4):1018-1031
A cell line ZBE3 isolated from a continuous cell culture derived from zebrafish Danio rerio blastomeres by clonal growth was characterized. ZBE3 cells had been subcultured for >120 passages since the initial primary culture of the blastomeres. The ZBE3 cells grow stably at temperature from 20 to 32° C with an optimum temperature of 28° C in ESM2 or ESM4 medium with 15% foetal bovine serum (FBS). The optimum FBS concentration for ZBE3 cell growth ranged from 15 to 20%. Cytogenetical analysis indicated that the modal chromosome number of ZBE3 cells was 50, the same as the diploid chromosome number of D. rerio. Significant cytopathic effect was observed in ZBE3 cells after infection with redspotted grouper nervous necrosis virus, Singapore grouper iridovirus and grass carp reovirus, and the viral replication in the cells was confirmed by real‐time quantitative PCR and transmission electron microscopy, indicating the susceptibility of ZBE3 cells to the three fish viruses. After transfected with pEGFP‐N3 plasmid, ZBE3 cells showed a transfection efficiency of about 40% which was indicated by the percentage of cells expressing green fluorescence protein. The stable growth, susceptibility to fish viruses as well as high transfection efficiency make ZBE3 cells be a useful tool in transgenic manipulation, fish virus‐host cell interaction and immune response in fish. 相似文献
3.
Summary Six established cell lines have been generated from embryos ofDrosophila melanogaster homozygous for different X-linked mutations. Four of these mutants, confer hypersensitivity to chemical mutagens in larvae.
The cell lines derived from the two mutageninsensitive stocks, serve as controls in the analyses of DNA metabolism. One cell
line (UCD-Dm-mei-9-2) is uniquely identified by a strong hypersensitivity to ultraviolet radiation. Another (UCD-Dm-mus104-1) expresses an enzyme variant not found in the other lines. The population doubling time for these cultures varies between
24 and 47 h. Labeling indices of 24.4 to 37.5% were found. The duration of the S phase in one of the control cell lines is
estimated to be about 9 h. Karyotype stability was monitored for five lines over a period of about 1 y. In general these cultures
each, became hypotetraploid with a preferential loss of the Y and fourth chromosomes. DNA synthesis in two of the lines fails
to exhibit the pattern of sensitivity to mutagens or caffeine that is observed in the corresponding primary cultures.
In primary cultures three classes of cells can be identified by autoradiography. About 50% of the cells label at a moderate
rate, 20% do not label within the first 1.5 d of culture, and the remaining cells exhibit a burst of labeling shortly after
the cultures are initiated.
This research was supported by NIH Grants GM16298 and GM22221 and by DOE Contract AT(04-3)-34 PA 210. 相似文献
4.
Xavier Lery Jean-Louis Zeddam Joseph Giannotti Liliane Croizier Gilles Fediere Said Abol-Ela 《In vitro cellular & developmental biology. Animal》1995,31(11):836-839
Summary A cell line from the main insect pest of potatoes in tropical and subtropical areas,Phthorimaea operculella (Zeller), was obtained from embryoculture. These cells were cultured in Grace’s modified medium. The cell line, designated
ORS-Pop-93, had a heterogeneous population consisting of spherical and spindle cells with great capacity to adhere and a doubling
time of 40 h. They were subcultured for more than 60 passages. Their polypeptidic profile was different from profiles of other
lepidopteran cell lines. The cell line supports the multiplication of theAutographa californica nuclear polyhedrosis virus. 相似文献
5.
Eun-Ho Park Jae-Seong Lee Ae-Kyng Yi Hisami Etoh 《In vitro cellular & developmental biology. Plant》1989,25(11):987-994
Summary A cell line (ULF-23HU) from the fin of the central mudminnow (Umbra limi) was characterized and tested for its suitability to assess cytogenetic damages induced by chemicals in fish. Cells of this
line exhibit a fibroblastlike appearance and grew optimal at 25°C in, TC-199 medium containing 10% fetal bovine serum, but
slower growth continued down to 4°C, where they could be stored for prolonged periods. Seeding efficiency of ULF-23HU cells
on the plastic substratum was approximately 85% in the above culture medium at 25°C. They had a 32-h cell cycle time taken
up by a 20-h S period as determined by the autoradiographic analysis of the fraction of labeled mitosis. Cultures showed relatively
high mitotic index (0.84 to 2.35%) during exponential growth phase lasting about 7 d. Karyological analysis of the cells at
the different subculture passages revealed constant chromosome modal number of 23 consisting of metacentric or submetacentric
chromosomes, which were primarily similar to those of in vivo cells, with one additional chromosome. The spontaneous sister
chromatid exchange rate was 5.3 per metaphse. When ULF-23HU cells were exposed toN-nitroso-N-methylurea, a clastogen in the mammalian cells, dose-dependent increases both in sister chromatid exchanges and chromosome
aberrations were clearly detected. These results on the growth kinetics and cytogenetic characteristics offered the high possibility
of the use of this cell line as a suitable in vitro model for clastogenicity studies in fish.
This work was supported by grants from Korea Sciences and Engineering Foundation and Japanese Government Research Awards for
Foreign Specialists to E.-H. Park. 相似文献
6.
Four new cell lines, designated as NTU-LY-1 to -4, respectively, were established from the pupal tissues of Lymantria xylina Swinhoe (Lepidoptera: Lymantriidae). These cell lines have been cultured approximately 80 passages during 2 years in TNM-FH medium supplemented with 8% fetal bovine serum, at a constant temperature of 28 degrees C. Each line consists of three major morphological types: round cells, spindle-shaped cells, and giant cells. The characterization of these cell lines showed that they are different from previously established lines derived from related Lepidopteran species. All new lines were susceptible to the L. xylina multiple nucleopolyhedrovirus (LyxyMNPV) and appeared to have a good potential for studying this virus. 相似文献
7.
J A Kang C-YShadduck 《In vitro》1977,13(12):849-856
Experimental tumors developed in white Pekin ducks after intramuscular implantation of 20-methylcholanthrene. Cells derived from the primary tumor were adapted successfully to grow in vitro and have growth characteristics similar to that of established cell lines of mammalian origin. The cell density rises rapidly and the doubling time is approximately 17 hr. The duck cells have been cultured successfully for at least 80 passages in votro. The continuously cultured cells have the characteristic chromosome pattern of duck, and the DNA of the duck cell line hybridized with duck liver DNA. We believe we have established a continuous cell line of avian origin. Electron-microscopic examinations of the tumor cells and RNA-directed DNA polymerase of the cell-free supernate show no evidence of endogenous virus production. 相似文献
8.
Toshinori Ide Yoshiaki Tsuji Tsuyoshi Nakashima Sadahiko Ishibashi 《Experimental cell research》1984,150(2)
Normal human diploid cells, TIG-1, ceased to proliferate at about the 62 population doubling level (PDL). Transformed clones isolated from TIG-1 cells infected with wtSV40 and those with tsA900 SV40 cultured at 34 °C were subcultured up to about 80 PDL. When the culture temperature of tsA SV40-transformed cells was shifted from 34 to 39.5 °C at 51 PDL, the growth curve of these transformed cells changed to that of normal young cells. When shifted to 39.5 °C after 62 PDL, cells immediately reached the end of their proliferative lifespan even under such favourable conditions for growth as low cell density in fresh medium. Growth of wtSV40-transformed cells did not change markedly at either temperature. These findings suggest that the clock of aging progresses in transformed cells as in normal cells, around 62 PDL being the senescent state in both cases, and that T-antigen of the tsA mutant of SV40 supports the extension of the lifespan of human cells only at the permissive temperature. 相似文献
9.
Fayuan Wen Yong-an Zhang Liangjian Qu Huan Zhang Zhongqi Yang Qilian Qin Yuzhu Wang Wenhuan Wang 《In vitro cellular & developmental biology. Animal》2009,45(8):409-413
Two cell lines designated CAF-Clan I and CAF-Clan II have been established from embryos of Clostera anachoreta (Lepidoptera: Notodontidae) in TNM-FH medium containing 10% inactivated fetal bovine serum. CAF-Clan I consists of a mixture
of three cell types: spherical cells, spindle-shaped cells, and giant cells. Most of the cultured cells formed a suspension
in the medium and were subcultured more than 60 passages. CAF-Clan II mainly consists of spindle-shaped and spherical cells
which attached to the culture surface and have undergone more than 40 passages. The cell population doubling time at 27°C
of CAF-Clan I at passage 22 and CAF-Clan II at passage 24 was about 68.5 and 38.2 h, respectively. The chromosome number of
both cell lines at passage 15 varied from 62 to 100 in the majority of cells, though a few cells exceeded 260 (n = 30). DNA amplification fingerprinting–polymerase chain reaction analysis confirmed that the origination of the two cell
lines was C. anachoreta. The susceptibility of the cell lines to baculoviruses was tested. The results showed that CAF-Clan II was susceptible to
infection of Autographa californica nucleopolyhedrovirus (AcMNPV) and Ecotropis oblique nucleopolyhedrovirus (EoNPV). Occlusion bodies (OBs) production was 129 ± 4 OBs/cell and 124 ± 15 OBs/cell for AcMNPV and
EoNPV, respectively. CAF-Clan I was less susceptible to AcMNPV compared with CAF-Clan II, while non-permissive to EoNPV. 相似文献
10.
Arun M. Khurad Min-Juan Zhang Chanchal G. Deshmukh Ravindra S. Bahekar Ashish D. Tiple Chuan-Xi Zhang 《In vitro cellular & developmental biology. Animal》2009,45(8):414-419
A new continuous cell line from ovarian tissue of commercial variety “Kolar Gold” of silkworm, Bombyx mori, was established and designated as DZNU-Bm-12. The tissue was grown in MGM-448 insect cell culture medium supplemented with
10% fetal bovine serum (FBS) and 3% heat-inactivated B. mori hemolymph at 25 ± 1°C. The migration of partially attached small round refractive cells from the fragments of ovarioles began
from the beginning of explantation. The cells multiplied partially attached in the primary culture initially, and some of
them become freely suspended after 20 passages. The cells were adapted to MGM-448 and TNM-FH media each with 10% FBS and the
population doubling time of cell line was about 36 and 24 hr, respectively. The chromosome number was near diploid at initial
passages and slightly increased at 176th passage, but a few tetraploids and hexaploids were also observed. DNA profiles using
simple sequence repeat loci established the differences between DZNU-Bm-12 and DZNU-Bm-1 and most widely used Bm-5 and BmN
cell lines. The cell line was found susceptible to B. mori nucleopolyhedrovirus (BmNPV) with 85–90% of the cells harboring BmNPV and having an average of 3–17 OBs/infected cell. We
suggest the usefulness of this cell line in BmNPV-based baculoviral expression system and also for studying in vitro virus
replication. 相似文献
11.
T. Raja Swaminathan Wazir S. Lakra A. Gopalakrishnan V. S. Basheer B. Khushwaha K. A. Sajeela 《In vitro cellular & developmental biology. Animal》2010,46(8):647-656
A new cell line [pearlspot fin (PSF)] has been developed from caudal fin of Etroplus suratensis, a brackish/freshwater fish cultivated in India. The cell line was maintained in Leibovitz’s L-15 supplemented with 10% fetal
bovine serum (FBS). The PSF cell line consisted predominantly of epithelial-like cells. The cells were able to grow at temperatures
between 25°C and 32°C with optimum temperature of 28°C. The growth rate of PSF cells increased as the FBS proportion increased
from 2% to 20% at 28°C with optimum growth at the concentration of 10% FBS. One marine fish virus (fish nodavirus) was tested
on this cell line and found not susceptible. After confluency, the cells were subcultured with a split ratio of 1:2. The cells
showed epithelial-like morphology and reached confluency on the third d after subculture. Polymerase chain reaction amplification
of mitochondrial 16S rRNA and COI indicated identity of this cell line with those reported from this fish species, confirming
that the cell line was of pearlspot origin. The cells were successfully cryopreserved and revived at the tenth, 25th, and
35th passages. The bacterial extracellular products from Vibrio cholerae MTCC 3904 were found to be toxic to PSF. Karyotyping analysis indicated that the modal chromosome number was 48. 相似文献
12.
Q Hu L Yu R Chen Y Zhang Y Xie Q Liao 《In vitro cellular & developmental biology. Animal》2012,48(7):441-448
Endometrial cancer is a major problem for women but only a small number of comprehensively characterized cell models are available for studies. Here, we established a new cell line derived from a Stage IIIc(1) Grade 2 endometrial adenocarcinoma. The cell line, designated LHY-821, was characterized using growth curve, karyotyping, immunohistochemical staining, immunoblotting, drug sensitivity assay, invasion assay, and xenografting in nude mice. LHY-821 has a doubling time of about 46?h and a colony-forming efficiency of approximately 71?%. These cells expresse high levels of progesterone receptor but not estrogen receptor and are sensitive to medroxyprogesterone acetate (MPA). LHY-821 also expresses pan-cytokeratin, PTEN, p53, β-catenin, IGF-1, and IGF-2. In addition, karyotype analysis revealed that LHY-821 possessed a near diploid karyotype including 6q-, 10p-, Xq-, 13q+, 17p+, and Triplo-12. LHY-821 showed highly tumorigenicity in nude mice (100?%) and weak invasiveness. Chemosensitivity tests showed that LHY-821 was sensitive to both carboplatin and paclitaxel. LHY-821 is an immortalized cell line which had survived more than 80 serial passages; it may provide a novel tool to study the molecular mechanism and potential treatment for endometrial cancer. 相似文献
13.
Lepidopteran heat-tolerant (ht) cell lines have been obtained with sf-9, sf-21 and several Bombyx cells. They have a distinct karyotype, membrane lipid composition, morphology and growth kinetics from the parental cell
lines. In this paper, we report the development of ht cell lines from other insect species and examination of their growth
characteristics and virus susceptibility. Adaptation of cell lines sf-9, BTI-TN-5B1-4 (High5) and BTI-TN-MG1 (MG1) to 33°C
and 35°C was carried out by shifting the culture temperature between 28°C and higher temperatures by a gradual stepwise increase
in temperature. The process of adaption to a higher culture temperature was accomplished over a period of 2 months. The cell
lines with the temperature adaption were designated as sf9-ht33, sf9-ht35, High5-ht33, High5-ht35, MG1-ht33, MG1-ht35. These
cell lines have been subcultured over 70 passages. Adaption to high temperatures was confirmed by a constant population doubling
time with individual cell lines. The population doubling time of heat adapted cell lines were 1–4 h less than these of parental
cell lines. Cell shapes did not show obvious change, however, the cell size of sf9-ht cells was enlarged and those of High5
and MG1 ht cells were reduced after heat adaption. When the cell lines were infected with Autographa californica nuclear polyhedrosis virus (AcMNPV) at 28°C, 33°C, 35°C and 37°C, production of budded virus and occlusion bodies in each
cell line was optimum at its own adapted temperature. 相似文献
14.
Herpes simplex virus DNA in transformed cells: sequence complexity in five hamster cell lines and one derived hamster tumor. 总被引:11,自引:7,他引:4
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Analyses of the hybridization kinetics of labeled herpes simplex virus 2 (HSV-2) DNA with DNA from five hamster cell lines transformed by UV light-irradiated HSV-2 revealed the following. (i) Viral DNA sequences were detected in all five cell lines tested. (ii) None of the cell lines contained the full complement of HSV-2 DNA. (iii) The amount of viral DNA present in the cells varied in different transformed cell lines and ranged from 8 to 32% of the HSV-2 DNA genome in 1 to 3 copies/cell. (iv) Two parallel passages of the same cell line (333-2-29) differed in the amount of viral DNA they contained. We also compared the viral DNA sequences present in (i) one transformed cell line (333-8-9) propagated serially in culture for 80 passages, (ii) a tumor produced by inoculation of a newborn hamster with the 333-8-9 cells, and (iii) a cell line derived from a hamster tumor as above and propagated in culture for 32 passages. The results show that viral DNA present in the hamster tumor and in the cells derived from the tumor had a lower sequence complexity than that present in the original serially passaged 333-8-9 cell line. 相似文献
15.
Trisyono A Goodman CL Grasela JJ McIntosh AH Chippendale GM 《In vitro cellular & developmental biology. Animal》2000,36(6):400-404
Summary A cell line derived from embryonic tissues of the European corn borer, Ostrinia nubilalis (UMC-OnE), was established in EX-CELL 401 medium containing 10% fetal bovine serum. The cells grew in suspension, and were
mainly spherical in shape. The cell doubling times at the 17th and 79th passages were 56 and 36 h, respectively. DNA amplification
fingerprinting showed that the DNA profile of the OnE cell line was different from that of the southwestern corn borer, Diatraea grandiosella (UMC-DgE), and that of the cotton bollworm, Helicoverpa zea (BCIRL-HZ-AM1). The OnE cell line was responsive to treatments of 20-hydroxyecdysone and the ecdysone agonists, methoxyfenozide
(RH-2485) and tebufenozide (RH-5992). These compounds caused similar effects on the cells, which included cell clumping and
decreased cell proliferation. The clumps were observed on the third day of incubation, and became larger after 7 d of incubation.
After 168 h of incubation, methoxyfenozide and tebufenozide were 35 and 11 times more effective, respectively, in inhibiting
proliferation of the OnE cells than was 20-hydroxyecdysone. 相似文献
16.
Yuanan Lu Vivek R. Nerurkar Alonso A. Aguirre Thierry M. Work George H. Balazs Richard Yanagihara 《In vitro cellular & developmental biology. Animal》1999,35(7):389-393
Summary Thirteen cell lines were established and characterized from brain, kidney, lung, spleen, heart, liver, gall bladder, urinary
bladder, pancreas, testis, skin, and periorbital and tumor tissues of an immature male green turtle (Chelonia mydas) with fibropapillomas. Cell lines were optimally maintained at 30° C in RPMI 1640 medium supplemented with 10% fetal bovine
serum. Propagation of the turtle cell lines was serum dependent, and plating efficiencies ranged from 13 to 37%. The cell
lines, which have been subcultivated more than 20 times, had a doubling time of approximately 30 to 36 h. When tested for
their sensitivity to several fish viruses, most of the cell lines were susceptible to a rhabdovirus, spring viremia carp virus,
but refractory to channel catfish virus (a herpesvirus), infectious pancreatic necrosis virus (a birnavirus), and two other
fish rhabdoviruses, infectious hematopoietic necrosis virus and viral hemorrhagic septicemia virus. During in vitro subcultivation,
tumor-like cell aggregates appeared in cell lines derived from lungs, testis, and periorbital and tumor tissues, and small,
naked intranuclear virus particles were detected by thin-section electron microscopy. These cell lines are currently being
used in attempts to isolate the putative etiologic virus of green turtle fibropapilloma. 相似文献
17.
18.
Atsushi Tsuboi Mieko Matsui Isamu Hayata Takehiko Tsuchiya 《In vitro cellular & developmental biology. Plant》1980,16(7):600-608
Summary The establishment of mouse tumor cell lines capable of proliferating in vivo and in suspension culture was undertaken. The
MM-46 tumor line, initiated from primary mammary carcinoma arising in a C3H/He mouse, was maintained for over 100 generations
in the peritoneal cavities of syngenic mice. At the 50th generation of the tumor suspension, cultures were initiated. The
established cell lines, designated TMT-1 and TMT-2, were characterized in vitro and in vivo. The morphological finding indicated
that TMT-1 and TMT-2 cells from mice closely resembled the MM-46 tumor cells. The oncogenic potential of the cultured cells
was comparable to that of the original ascites tumor. The population doubling time of TMT-1 and TMT-2 cell lines was about
12 hr in mice, whereas the population doubling time of both cell lines lengthened to 20 hr in suspension culture. The increase
of doubling time in culture was due to the prolongation of the G1 period. The cell lines, TMT-1 and TMT-2, whether from culture or mice, possessed colony forming ability in soft agar medium.
The colony forming ability of the cells decreased gradually through in vivo passages but it recovered upon recultivation of
the cells from mouse to culture. Chromosome analysis and cytotoxicity test by anti-MM antiserum indicated that TMT-1 and TMT-2
cell lines closely resembled and had been derived from MM-46 tumor line. Therefore, it is possible to assay cell survival
in vitro after in vivo experiments on these cells. 相似文献
19.
Belloncik S Petcharawan O Couillard M Charpentier G Larue B Guardado H Chareonsak S Imanishi S 《In vitro cellular & developmental biology. Animal》2007,43(7):245-254
The corn borer, Ostrinia nubilalis, is a very important pest in different countries, and the in vitro system of the insect could be a useful tool for isolation
and characterization of the pathogens and physiological responses of the insect. In this context, a cell line was derived
from the hemocytes of the European corn borer and was named AFKM-On-H for, respectively, O. nubilalis, Armand Frappier, King Mongkut Institutes, and Hemocytes. This cell line was initiated and maintained in Ex-Cell 400 medium
supplemented with 10% heat-inactivated fetal bovine serum. The cells, mostly spherical in shape, not firmly attached to the
plastic culture flasks, were passaged up to 200 times by repeated gentle pipetting of the cells. The doubling times at the
80th and 125th passages at 28°C and at the 122th and 169th passages at 25°C were 40, 29, 35, and 34 h, respectively. The AFKM-On-H
cell line was further characterized by the morphology, karyotype, random amplified polymorphic DNA analysis, and isozyme profiles.
Susceptibility of the cell line to cytoplasmic polyhedrosis viruses (CPV) Euxoa scandens (EsCPV), Dendrolimus punctatus (DpCPV), and Choristoneura fumiferana (CfCPV); nuclear polyhedrosis viruses [Autographa californica (AcMNPV) wild type and recombinant, Antherea yammamai (AnyaNPV)]; and Chilo iridescent virus was demonstrated. Relative sensitivities of the cell line to Bacillus thuringiensis and Metarhizium anisopliae toxins and effects of the molting hormone 20-hydroxyecdysone on this new hemocyte cell line were characterized. 相似文献
20.
Two new fish cell lines were established from skin (LWSK) and fin (LWFN) of leopard wrasse Macropharyngodon geoffroy. These cells grew optimally at 25° C in Leibovitz‐15 medium supplemented with 10% foetal bovine serum. Proliferation of M. geoffroy cells remained serum dependent up to cell passage 16, and cell‐plating efficiency ranged from 12 to 16%. Karyotypic analysis of these new cell lines at cell passage 8 indicated that both cell lines remained diploid with a peak chromosomal count of 144. PCR amplification of 16S mitochondrial DNA and the subsequent analysis confirmed that these cell lines were indeed derived from M. geoffroy. Results of viral challenge assays revealed that both LWSK and LWFN shared patterns of viral susceptibility similar to that of six fish viruses tested: LWSK and LWFN cells were highly permissive to channel catfish virus, spring viremia carp virus and snakehead rhabdovirus with high‐yield virus production ranging from 107·18±0·17 to 108·37±0·16 TCID50 ml?1 (mean ± s.d .). These newly established cell lines would be useful in attempts to isolate and study aquatic viruses, particularly the viral aetiology of green turtle fibropapilloma as M. geoffroy is known to be one of the common cleaner fish of green sea turtles. 相似文献