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1.
Fish cell lines: Establishment and characterization of nine cell lines from salmonids 总被引:3,自引:0,他引:3
C. N. Lannan J. R. Winton J. L. Fryer 《In vitro cellular & developmental biology. Plant》1984,20(9):671-676
Summary Nine permanent cell lines have been established from five species of salmonids native to America's Pacific Northwest. With
the exception of a hepatoma from an adult trout, the lines were derived from normal tissues of embryonic or juvenile fish.
Cells were routinely grown in Eagle's minimum essential medium with 10% fetal bovine serum. Optimum growth temperatures for
these lines ranged from 21 to 24°C. All survived storage for at least 1 yr at −65°C and at least 5 yr in liquid nitrogen.
Six of the lines were demonstrably free of any microbial contamination but mycoplasmas were found in three. Eight of the lines
were heteroploid. The morphology of only one was fibroblastic. All the lines effectively replicated one or more of the common
salmonid viruses. Isozyme patterns were consistent with those of the species of origin. These cell lines have significant
application in fish virology.
This work is a result of research sponsored in part by the Oregon State University Sea Grant College Program supported by
NOAA Office of Sea Grant, U.S. Department of Commerce, under Grant NA79AA-D-0016 and by the Oregon Department of Fish and
Wildlife under PL-89304 Anadromous Fish Act and is Oregon Agricultural Experiment Station Technical Paper 6857. 相似文献
2.
Yuanan Lu C. N. Lannan J. S. Rohovec J. L. Fryer 《In vitro cellular & developmental biology. Plant》1990,26(3):275-279
Summary Three new cell lines were established from tissues of the grass carp,Ctenopharyngodon idella. Derived from the fin, snout, and swim bladder of two apparently healthy diploid fry, these cell lines have been designated
GCF, GCS-2, and GCSB, respectively. The cells grew at temperatures between 24° and 36° C with optimal growth at 32° C and
have been subcultured more than 50 times since their initiation in August 1986. Two of the lines remained diploid or pseudodiploid
after 38 passages. The cells were tested for microbial contamination, and plating efficiencies were determined. The three
cell lines were sensitive toRhabdovirus carpio (RVC), infectious hematopoietic necrosis virus (IHNV), golden shiner virus (GSV), chum salmon virus (CSV), and infectious
pancreatic necrosis virus serotype VR299 IPNV). They were refractory to channel catfish virus (CCV), channel catfish reovirus
(CRV), chinook salmon paramyxovirus (CSP), and an Ab serotype of IPNV.
This work is a result of research sponsored by the Oregon State University Sea Grant College Program supported by NOAA Office
of Sea Grant, U.S. Department of Commerce, under grant NA85AA-D-5G-095, and was undertaken while the first author was on leave
from the Department of Fisheries, Huazhong Agricultural University, Wuhan, China. Salary support was provided by the People's
Republic of China. Oregon Agricultural Experiment Station Technical Paper 8952. 相似文献
3.
B. L. Nicholson D. J. Danner J -L. Wu 《In vitro cellular & developmental biology. Plant》1987,23(3):199-204
Summary Three new continuous cell lines were established from two species of marine fishes economically important in Asia. Perch (Lateolabrax japonicus) heart (PH), perch liver (PL), and grouper (Epinephelus amblycephalus) kidney (GK) lines were established in Eagle’s minimum essential medium with 10% fetal bovine serum and have been subcultured
over 120 times. The optimum growth temperature was 25°C for the PK and GK lines and 30° C for the PH line. The modal chromosome
numbers for each cell line are: PH (49), PK (50), and GK (65). None of the lines was susceptible to the rhabdovirus infectious
hematopoietic necrosis virus (IHNV) or channel catfish herpesvirus (CCV); however, all three cell lines were susceptible to
a variety of fish birnaviruses, including infectious pancreatic necrosis virus (IPNV), the EVE eel virus, and newly isolated
birnaviruses from a variety of fish and shellfish in Taiwan.
This research was funded by National Science Foundation grant INT-810447, the University of Main/University of New Hampshire
Sea Grant College Program, and the Maine Agriculture Experiment Station publication no. 1165. 相似文献
4.
Fish cell lines: Two new cell lines derived from explants of trunk musculature ofCynoscion arenarius
B. L. Middlebrooks D. L. Stout R. D. Ellender S. Safford 《In vitro cellular & developmental biology. Plant》1981,17(5):427-430
Summary Two cell lines were established from explants of trunk musculature of healthy, males sand seatrout,Cynoscion arenarius. One of the lines, designated CyA-1, has been carried through 150 subcultures during 6 yr. The other, designated CyA-2, has
been carried through 100 subcultures during 2 yr. Both lines grow well in L15 medium adjusted to 0.150M NaCl and supplemented with 10% fetal bovine serum. Optimal growth occurs at temperatures between 24 and 30°C. The species
of origin of both lines was confirmed by a cytotoxic antibody dye exclusion test. The karyotype of CyA-1 has not yet stabilized,
showing a modal chromosome number of 120 at Passage 9, 89 at Passage 63, and 79 at Passage 100. The karyotype of CyA-2 is
rather stable, with a modal chromosome number of 47 at Passage 1 and 49 at Passage 100. Chromosome morphology of CyA-2 is
homogeneous (small, acrocentric), whereas the chromosomes of CyA-1 show considerable size variation (with small chromosomes
possibly formed from fragmentation of original structures). Both lines were found to be free of bacterial or fungal contamination.
Both lines supported replication of lymphocystis virus strains isolated fromBairdiella chrysura (the silver perch) and fromMicropogon undulatus (the Atlantic croaker) but were refractory to 11 other viruses (4 from fish, 1 from amphibians, and 6 from mammals).
This study was supported in part by the National Oceanographic and Atmospheric Administration, Office of Sea Grant No. 04-3-158-58. 相似文献
5.
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
6.
Burt C. Lidgerding Stevan R. Phelps William B. Schill 《In vitro cellular & developmental biology. Plant》1984,20(3):167-171
Summary The electrophoretic mobilities of isozymes from 16 enzyme systems were determined for nine fish cell lines. The relative migration
of the malate dehydrogenase and 6-phosphoglucose dehydrogenase isozymes could be used together to distinguish between seven
of the fish cell lines. Two cyprinid cell lines could not be distinguished from each other but were readily separated from
the six noncyprinid lines and the one other line of cyprinid origin. 相似文献
7.
S. Wright-Perkins M. R. Daniel C. Walker 《In vitro cellular & developmental biology. Animal》1994,30(7):450-459
Summary This study investigates the characteristics of two human cell lines—1PT and 1PT VARIANT A—both derived from the same histologically
undifferentiated, neuroendocrine positive, non-small cell lung carcinoma (NSCLC) and capable of growth in unsupplemented serum-free
minimum essential medium. In stationary culture, the cells of both lines grew both attached to a plastic substratum and in
suspension; the 1PT VARIANT A line formed three-dimensional clusters of loosely adherent cells. The cell lines differed in
their DNA content, the 1PT having 1.44 times and the 1PT VARIANT A having 2.39 times the normal human diploid DNA content.
Chromosome counts supported this observation, the ploidy of the 1PT and VARIANT A lines being 1.11 and 1.64, respectively.
On transmission electron microscopy the cells of both lines had dense core granules and immature desmosomes, whereas only
the 1PT VARIANT A line had mucin granules. Both lines formed, in nude mice, tumors that, like the original tumor from which
they were derived, were histologically undifferentiated and showed local invasion. The original tumor and both lines had demonstrable
neuroendocrine markers. Cytokeratins were apparent in the tumor but not the cell lines, and neurofilaments were present in
the cell lines only. Staining for epithelial membrane antigen, neural cell adhesion molecule, and desmoplakin differentiated
between the two lines. These lines provide a useful model for the investigation of the biology of the neuroendocrine positive
subgroup of NSCLC, which is clinically important because of the possible responsiveness of these tumors to chemotherapy. 相似文献
8.
Jun-Ichiro Komura Hiroshi Mitani Akihiro Shima 《In vitro cellular & developmental biology. Plant》1988,24(4):294-298
Summary Two fibroblast-like cell lines were obtained from fins of adults of the medaka,Oryzias latipes, and serially cultured at 27° C. One cell line, which was derived from a fish of the orange-red variety, reached to population
doubling level (PDL) 434 on Day 840 by 120 passages. The other, which was derived from a fish of the inbred strain HB32C,
reached to PDL 294 on Day 551 by 80 passages. Any symptom of crisis was not detected. The cell lines were named OL-17 and
OL-32, respectively. Population doubling time was 29 h for OL-17 cells, and 32 h for OL-32 cells. Density-dependent inhibition
of growth was clear in OL-32 cells, but not so obvious in OL-17 cells. Modal chromosome number of OL-17 cells was 50, and
that of OL-32 cells was 47 (2N=48). Plating efficiency of OL-17 cells was about 10%, whereas that of OL-32 cells was about
5%. The use of conditioned medium (80% concentration) increased the plating efficiency of OL-32 cells to more than 25%. 相似文献
9.
A proteomics approach to identifying fish cell lines 总被引:2,自引:0,他引:2
Fish cell lines are relatively easy to culture and most have simple growth requirements that make cross contamination a potential problem. Cell line contamination is not an uncommon incident in laboratories handling more than one cell line and many reports have been made on cross contamination of mammalian cell lines. Although problems of misidentification and cross-contamination of fish cell lines have rarely been reported, these are issues of concern for cell culturists that can make scientific results and their reproducibility unreliable. Proper identification of cell lines is thus crucial and protocols for routine and rapid screening are preferred. Cytogenetic evaluation, DNA fingerprinting, microsatellite analysis and PCR methods have been published for inter-species identification of many cell lines, but discerning intra-species contamination has been challenging. More complex DNA fingerprinting and hybridization techniques coupled with isoenzyme analysis have been developed to discriminate intra-species contamination, however, these require complex and time consuming procedures to enable cell identification thus are difficult to apply for routine use. A simple proteomic approach has been made to identify several fish cell lines derived from tissues of the same or differing species. Protein expression signatures (PES) of the evaluated fish cell lines have been developed using 2-DE and image analysis. A higher degree of concordance was seen among cell lines derived from rainbow trout, than from other fish species. Similar concordance was seen in cells derived from the same tissues than from other tissues within the same species. These profiles have been saved in an electronic databank and could be made available to be used for discerning the origins of the various cell lines evaluated. This proteomic approach could thus serve as an additional, valuable and reliable technique for the identification of fish cell lines. 相似文献
10.
Summary A cell line was established from the ovaries of a healthy, juvenile channel catfish,Ictalurus punctatus. These cells, designated CCO, have been passed 130 times during 3 years. The cells grow well in Eagle's MEM-10 at a temperature
of 30°C. Species of origin of the cells was confirmed by a cytotoxic dye exclusion test. The cells were found to be free of
bacterial and fungal contamination. A study of chromosome preparations indicated that the karyotype is still in a state of
flux. The CCO line replicated channel catfish virus but was refractory to 12 other viruses, 4 from fish, 1 from birds, and
7 from mammals.
This research was supported by the Southeastern Cooperative Fish Disease Project, Sport Fish Restoration Funds and Regional
Research S-83 Funds. 相似文献
11.
Summary Microarray technology has burgeoned over the past few years from nucleic acid-based arrays to tissue microarrays (TMAs). This
study aimed to develop a technique to incorporate cell lines into an array and to demonstrate the usefulness of this technique
by performing immunohistochemistry for β-catenin. Cell suspensions were prepared from 23 tumor cell lines. These were fixed
in formalin, suspended in agar, and embedded in paraffin to produce a cell block. A “tissue microarrayer” was used to remove
triplicate, 0.6 mm-cores from each cell block and to transfer these into a recipient paraffin block at precise coordinates.
Immunohistochemistry was used to identify cell lines positive for β-catenin. Cultured cells were successfully incorporated
into the microarray, with preservation of cell architecture and even distribution of cells within each core. A total of 18
of 69 cores (26%) were lost in processing. A total of 16 of 23 cell lines were identified as positive for membrane and cytoplasmic
β-catenin, and 6 of 23 were negative. Only one cell line was unscorable because of complete core loss. We have developed a
“cell microarray” technique for analyzing antigen expression by immunohistochemistry in multiple cell lines in a single expriment.
This novel application of microarrays permits high-throughput, cost-efficient analysis, with the potential to rapidly identify
markers with potential diagnostic and therapeutic implications in human disease. 相似文献
12.
Rui Yan Mark Ottenbreit Bharati Hukku Michael Mally Sharong Chou Joseph Kaplan 《In vitro cellular & developmental biology. Animal》1996,32(10):656-662
Summary Methods for monitoring cell line identification and authentication include species-specific immunofluorescence, isoenzyme
phenotyping, chromosome analysis, and DNA fingerprinting. Most previous studies of DNA fingerprinting of cell lines have used
restriction fragment length polymorphism analysis. In this study, we examined the utility of an alternative and simpler method
of cell line DNA fingerprinting—polymerase chain reaction (PCR) amplification of fragment length polymorphisms. Fourteen human
cell lines previously found by other methods to be either related or disparate were subjected to DNA fingerprinting by PCR
amplification of selected fragment length polymorphism loci. Cell identification patterns by this method were concordant with
those obtained by isoenzyme phenotyping and restriction fragment length polymorphism-DNA fingerprinting, and were reproducible
within and between assays on different DNA extracts of the same cell line. High precision was achieved with electrophoretic
separation of amplified DNA products on high resolution agarose or polyacrylamide gels, and with fragment length polymorphism
(FLP) loci-specific “allelic ladders” to identify individual FLP alleles. Determination of the composite fingerprint of a
cell line at six appropriately chosen fragment length polymorphism loci should achieve a minimum discrimination power of 0.999.
The ability of PCR-based fragment length polymorphism DNA fingerprinting to precisely and accurately identify the alleles
of different human cell lines at multiple polymorphic fragment length polymorphism loci demonstrates the feasibility of developing
a cell line DNA fingerprint reference database as a powerful additional tool for future cell line identification and authentication. 相似文献
13.
DNA fingerprinting is an important new development for the authentication of cell lines. Multilocus methods such as those
developed by Alec Jeffreys provide information on a wide range of genetic loci throughout the human genome and thus give a
useful genetic “snap-shot” of a cell culture. Our work has shown that Jeffreys multilocus fingerprinting method can be applied
to cell lines from a wide range of animals including reptiles, birds, fish and diverse mammals. It can also differentiate
very closely related cell lines including those from the same mouse strain. Routine fingerprint analysis has enabled an unprecedented
level of confidence in the consistency of cell stocks. Our results demonstrate that this straightforward method represents
a powerful and readily interpreted system for cell authentication and exclusion of cross-contamination. 相似文献
14.
G. Contreras R. Bather J. Furesz 《In vitro cellular & developmental biology. Plant》1985,21(11):649-652
Summary Studies on the tumorigenicity of Vero kidney cells ofCercopithecus aethiops monkey origin were extended to various passage levels of BSC-1 aneuploid cells and to low passage CV-1 diploid cells (derived
also fromC. aethiops monkey kidney). It was found that BSC-1 cells—like Vero cells—showed increased tumorigenicity with increasing passage level
in antitymocyte globulins (ATG) treated newborn rats and in nude mice. Cells passaged over 250 times in cultures formed invasive
adenocarcinomas in newborn rats. Their malignant tumor growth was further demonstrated around the 500 passage level when tumor
metastases were detected in the lungs of four of the 14 inoculated rats. Vero cells induced such lung metastases in rats already
at passage 227. CV-1 diploid cells at low passage level produced small nodules of epithelioid cells in newborn rats at 6th
day after inoculation that had disappeared by the 21st day, and caused no local invasion nor lung metastasis. In vitro tumorigenicity
tests on BSC-1 and CV-1 cells, using chick embryo skin, human muscle and colony formation in agarose, confirmed the animal
test results.
The results of this study indicate that BSC-1 and Vero cell lines at low and high passage levels may prove to be useful tools
to study the molecular basis of malignancy.
Editor's Statement In this paper Contreras et al. document the increased malignancy of commonly-used monkey cell lines upon
long-term culture. These observations have implications for the use of these cell lines in studies of cancer cell biology,
as well as the use of these lines for the production of biologicals. David W. Barnes 相似文献
15.
Gary K. Ostrander James B. Blair Beverly A. Stark Garry M. Marley Wesley D. Bales Robert W. Veltri David E. Hinton Mark Okihiro Lisa S. Ortego William E. Hawkins 《In vitro cellular & developmental biology. Animal》1995,31(5):367-378
Summary Long-term primary cultures of epithelial cells from rainbow trout (Oncorhynchus mykiss) liver have been established. Nearly homogenous (>97%) populations of hepatocytes were placed into primary culture and remained
viable and proliferative for at least 70 d. In addition to hepatocytes, proliferative biliary cells persisted in the cultures
for at least 30 d. Finally, a third type of epithelial cell, which we have termed a “spindle cell,” consistently appeared
and proliferated to confluence in these cultures. The confluent cultures of spindle cells were successfully subcultured and
passaged.
The initial behavior, growth, and optimization of serum and media requirements for these cells is described. All three cell
types proliferated as measured by thymidine incorporation, autoradiography, proliferating cellular nuclear antigen analysis,
and propidium iodine staining. Further efforts to characterize the cells included western blotting and immunohistochemical
staining with antibodies to cytokeratins previously reported in fish liver. From these data, it appears that all three cell
populations are epithelial in nature. Furthermore, significant changes in actin organization, often indicative of transformation
or pluripotent cells, were observed with increased time in primary culture. 相似文献
16.
E. González-Reimers F. Santolaria-Fernández A. Moreno-García N. Batista-López F. Rodríguez-Moreno 《International Journal of Anthropology》1993,8(1):21-25
Harris lines represent episodes of growth arrest followed by recovery, thus reflecting episodes of mismatched imbalance between
energy intake and energy expenditure. Consumption of ethanol during the growth period — formerly extended in our wine-producing
rural areas- may lead to such a situation, due at least in part to the energy-wasteful MEOS-linked ethanol metabolism. Based
on these facts, in the present study we have analysed the relationship between ethanol intake during growth period and the
presence of Harris lines in the right tibia in 100 consecutive adult patients admitted to our hospitalization unit. Early
ethanol consumption was strongly related with the presence of two or more, and three or more Harris lines, so that ethanol
consumption should be considered in the differential diagnosis of Harris lines, and conversely, the presence of Harris lines
point to ethanol consumption during growth period. This association was independent from coexisting starvation and/or illness. 相似文献
17.
鱼类细胞培养尽管起步较晚,但截至目前已有近280余株不同的鱼类细胞系相继建立起来,在生理学、病毒学、毒理学、肿瘤及基因工程等多个领域发挥重要作用。主要对鱼类细胞系在病毒学研究中的最新应用,并结合作者自身的研究,尤其侧重于鱼类病毒分离、重要功能基因鉴定、抗病毒免疫和病毒致病机理研究等方面的进展作一概述。 相似文献
18.
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
19.
A fish expression vector, FRM, was constructed by fusing the carp β-actin promoter and first intron to the ocean pout antifreeze
protein terminator and putative boundary element. Mutant forms of the green fluorescent protein (GFP) were engineered into
this vector, and the resultant series of vectors, FRMwg, FRM3wg (green GFP), and FRM2bl (blue GFP), were used to make transgenic
zebra fish. After microinjection of either supercoiled or linearized DNA into one-celled eggs, GFP-expressing cells could
be monitored by fluorescence microscopy commencing with the midblastula transition and continuing through embryogenesis. From
adult fish, which retained scorable GFP either as patches or as a uniform fluorescence, 11 green and 1 blue GFP-expressing
lines of zebra fish have been established. Expression of GFP was nearly ubiquitous and similar among all of these lines. Embryonic
expression could be scored at 15 to 30 hours postfertilization and was seen throughout the embryo with the exceptions of the
yolk, red blood cells, and in some lines, portions of the head. Adult expression was in a majority of tissues (e.g., muscle,
brain, intestine, and heart, but not red blood cells). The notable difference between lines was that fluorescent eggs were
scorable in seven of the lines. Adult homozygotes from a different subset of eight lines could be selected by the relative
intensity of the GFP marking when compared with that in sibling heterozygotes. All 12 lines contain apparent single locus,
multicopy, tandem integrations (1.5–100 copies per cell) of the transgenic DNA. The fish expression vector FRM could be used
to drive nearly ubiquitous and strong expression of gene products other than GFP. The GFP expression vectors, FRMwg, FRM2wg,
FRM3wg, and FRM2bl, may be useful for optimization of transgenesis and as a comarker. GFP-expressing zebra fish lines could
facilitate experimental analysis of chimerism and in vivo gene targeting.
Received May 18, 1999; accepted August 26, 1999. 相似文献
20.
The differential cytotoxic effects of cadmium on fish and mammalian epithelial cells in established and primary culture were assessed by looking at the reduction of the colony-forming ability and reduction in the extent of growth. The influence of medium composition on the toxicity of cadmium was also studied using serum-free and serum-containing media. The experiments using immortalized cell lines showed that mammalian cells were more sensitive than fish cells to cadmium. Both cell types were grown at the same serum concentration. However, using the normal primary system, human epithelial tissue explants showed less sensitivity to cadmium than did similar cultures from rainbow trout. It is likely that cellular mechanisms of cadmium resistance in the different cell types are responsible for these effects. As expected, cadmium proved to be more toxic when tested in serum-free medium. With fish skin primary cultures, reduction of cell numbers was observed at concentrations as low as 5 mol/L in serum-free medium compared to 100 mol/L in serum-containing medium. This was found to be due to the reduction in the activity of free cadmium ions, caused by the presence of serum in the medium. It is concluded that serum-free formulations are probably preferable when testing pollutants in vitro. The results highlight the importance of establishing species-specific pollution tests and standardizing variables.In summary, the results show species and culture media differences in cadmium toxicity and suggest that caution is required when extrapolating results for pollutant effects from one in vitro system to another.Abbreviations CE
colony-forming efficiency
- EPC
epithelioma papulosum cyprini
- KGM
Clonetics Keratinocyte Growth Medium 相似文献