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1.
The zebrafish has become a popular model for studies of vertebrate development and toxicology. However,in vitro approaches utilizing this organism have not been fully exploited due to the absence of suitable cell culture systems. Previously, we developed methods for the culture of cells derived from zebrafish blastula-stage embryos. One of these cultures, ZEM-2, was derived in a complex medium containing trout embryo extract, trout serum and medium conditioned by buffalo rat liver cells. In this study we describe a zebrafish embryo cell line, ZEM-2A, derived from ZEM-2 following selection for growth in a simplified medium. Optimal growth of ZEM-2A cells is attained in nutrient medium supplemented with 5% fetal bovine serum. 相似文献
2.
Summary A continuous line of epithelioid cells was established from explant skin tissues of the green sea turtle,Chelonia mydas. These cells, designated GTS, have been subcultured more than 60 times in commercially available mammalian cell culture medium supplemented with 5% bovine calf serum. Of those temperatures tested, optimal growth was achieved at 30°C although replication occurred between 16 and 37°C. These cells may be held as monolayers at 8°C or stored frozen in growth medium containing 10% dimethylsulfoxide at −70 or −196°C. The modal number of 55 chromosomes per cell is in agreement with the heterogametic female diploid number of this species. The GTS line represents the first established culture of normal epithelioid skin cells to be reported for a poikilothermic species. 相似文献
3.
Qimeng Shi Yuhang Zhuang Tingting Hu Chuncheng Lu Xinru Wang Hongyu Huang Guizhen Du 《Journal of biochemical and molecular toxicology》2019,33(5)
Triclocarban (TCC), which is used as an antimicrobial agent in personal care products, has been widely detected in aquatic ecosystems. However, the consequence of TCC exposure on embryo development is still elusive. Here, by using zebrafish embryos, we aimed to understand the developmental defects caused by TCC exposure. After exposure to 0.3, 30, and 300 μg/L TCC from 4‐hour postfertilization (hpf) to 120 hpf, we observed that TCC exposure significantly increased the mortality and malformation, delayed hatching, and reduced body length. Exposure to TCC also affected the heart rate and expressions of cardiac development–related genes in zebrafish embryos. In addition, TCC exposure altered the expressions of the genes involved in hormonal pathways, indicating its endocrine disrupting effects. In sum, our data highlight the impact of TCC on embryo development and its interference with the hormone system of zebrafish. 相似文献
4.
Characterization of a cell line (SW756) derived from a human squamous carcinoma of the uterine cervix 总被引:6,自引:0,他引:6
Ralph S. Freedman James M. Bowen Albert Leibovitz Sen Pathak Michael J. Siciliano Harry S. Gallager Beppino C. Giovanella 《In vitro cellular & developmental biology. Plant》1982,18(8):719-726
Summary An established cell line, SW756, derived from a primary squamous carcinoma of the uterine cervix is described by its morphology,
ultrastructure, karyotype, genetic signature analysis, HLA typing, and tumorigenesis in the nude mouse. Cultured cells obtained
from the SW756 derived nude mouse tumor also were studied for chromosome and isozyme markers. The original tumor was poorly
differentiated carcinoma with minimal keratinization and is compared with that occurring in the nude mouse after the cultured
cells were inoculated. The nude mouse tumor showed similar histological features, but better differentiation than the original
tumor. Karyotype analysis of SW756 demonstrated a hyperdiploid stem line number and several marker chromosomes (MI-M6). No
HeLa marker chromosomes were identified. The isozyme pattern for SW756 reported by others has been confirmed. The unique chromosome
and isozyme features have been identified repeatedly in the cultured cells and, most importantly, in the post nude mouse culture.
We recommend SW756 as a defined human tumorigenic cell line derived from a primary squamous carcinoma of the uterine cervix.
This investigation was supported in part by Public Health Research Grant CA-06294 from the National Cancer Institute, Department
of Health and Human Services. 相似文献
5.
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
6.
一株棕尾别麻蝇胚胎细胞系的建立及其特性分析 总被引:1,自引:0,他引:1
双翅目昆虫细胞系广泛应用于遗传学、发育生物学、分子生物学、人和动物体病原学以及昆虫抗微生物肽的研究。本研究建立了一株新的棕尾别麻蝇Sarcophaga peregrina胚胎细胞系。该细胞系的原代培养始于2008年11月17日, 取材于棕尾别麻蝇晚期胚胎组织, 在Shields & Sang M3昆虫培养基中于28℃恒温培养, 在第26天进行第1次传代, 至今已历时21个月, 传代72次, 生长状态稳定, 被命名为Sp-E-HNU11。该细胞系的细胞形态主要呈梭形和近圆形, 杂以少量巨型细胞, 紧密贴壁生长。细胞群体倍增时间为42 h。染色体数目一般为10条或12条, 为二倍体或亚二倍体细胞系; 除一对颗粒状微型染色体外, 其他染色体呈短杆状。细胞系的β-萘酯酶和谷草转氨酶同工酶谱上分别显示出1条和3条酶带。随机引物扩增多态性 (random amplified polymorphic DNA, RAPD) 分析结果显示, 该细胞系与小菜蛾细胞系Px-E-HNU12、草地贪夜蛾细胞系IPLB-Sf-9和家蚕细胞系Bm-21E-HNU5呈现明显不同的带型特征。 Sp-E-HNU11细胞系的建立为昆虫抗微生物肽及其他相关的研究工作增添了新的研究工具和生产载体。 相似文献
7.
R. Rochford E. M. Dougherty D. E. Lynn 《In vitro cellular & developmental biology. Plant》1984,20(11):823-825
Summary A new cell line was developed from 3-d-old embryonated eggs of the cabbage looper,Trichoplusia ni, and has been designated IPLB-TN-R2. It contains a variety of morphological cell types, including myoblastlike, neuroblastlike, and epithelial-like cells. Chromosome
analysis revealed typical lepidopteran chromosomes. Isozyme characterization showed patterns similar to two other cabbage
looper cell lines (TN-368 and IAL-TND1) in the case of five enzymes but differed from these two lines for two other enzymes.
Virus infectivity tests revealed the line is highly susceptible toAutographa californica nuclear polyhedrosis virus, but no cytopathology was observed after inoculation with several other lepidopteran viruses. 相似文献
8.
Beryl Vedha Yesudhason Johnson Retnaraj Samuel Selvan Christyraj Mijithra Ganesan Karthikeyan Subbiahanadar Chelladurai Saravanakumar Venkatachalam Arun Ramalingam Johnson Benedict Vennila Devi Paulraj Jackson Durairaj Selvan Christyraj 《Cell biology international》2020,44(10):1968-1980
Zebrafish (Danio rerio), is a well‐established vertebrate animal model widely used in developmental biology and toxicological research. In the present study, foldscope is used as an innovative tool to study the developmental stages and toxicological analysis of the zebrafish embryos. Briefly, the developmental stages, such as zygote, cleavage, blastula, gastrula, segmentation, and pharyngula formation are observed and documented using simple foldscope. Toxicological parameters upon exposure to different concentration of ethanol extract of Curcuma longa and its lead compound, ar‐turmerone along with rhodamine B (bio‐coupler) on zebrafish embryos are analyzed upto 72 hr using foldscopes in live condition. The lethal endpoints, such as coagulation, lack of somite formation, non‐detachment of tail, and lack of heartbeat are clearly monitored and documented using foldscope. Bio‐evaluation of test compounds with the aid of foldscope confirms that the toxicity is directly proportional to the concentration. Our results conclude that, ethanol extract of C. longa, ar‐turmerone and rhodamine B exposed embryos remains healthy up to 96, 48, and 24 µg concentrations, respectively. Embryos exposed to higher concentrations become coagulated, however normal physiological active movement of tail lashing and heartbeat are evident in lower concentration exposed embryos. Except coagulation, no other abnormalities are observed and interestingly, the hatching ability is not delayed, when compared with the control embryos. It is confirmed that the test compounds are not highly toxic to zebrafish embryos. Hence it can be used for further analysis, especially for studying the neural‐regeneration and its neuronal development in zebrafish embryos. 相似文献
9.
Ting Tao Jinrong Peng 《遗传学报》2009,36(6):325-334
Liver is one of the largest internal organs in the body and its importance for metabolism, detoxification and homeostasis has been well established. In this review, we summarized recent progresses in studying liver initiation and development during embryogenesis using zebrafish as a model system. We mainly focused on topics related to the specification of hepatoblasts from endoderm, the formation and growth of liver bud, the differentiation of hepatocytes and bile duct cells from hepatoblasts, and finally the role of mesodermal signals in controlling liver development in zebrafish. 相似文献
10.
The zebrafish is a polular nonmammalian model for studies of neural development. We have derived cell cultures, initiated from blastula-stage zebrafish embryos, that differentiate in vitro into neurons and astrocytes. Cultures were initiated in basal nutrient medium supplemented with bovine insulin, trout serum, trout embryo extract and fetal bovine serum. After two weeks in culture the cells exhibited extensive neurite outgrowth and possessed elevated levels of acetylcholinesterase enzyme activity. Ultrastructural analysis revealed that the neurites possessed microtubules, synaptic vessicles and areas exhibiting growth cone morphology. The cultures expressed proteins recognized by antibodies to the neuronal and astrocyte-specific markers, neurofilament and glial fibrillary acidic protein (GFAP). Poly-D-lysine substrate stimulated neurite outgrowth in the cultures and inhibited the growth of nonneuronal cells. Medium conditioned by the buffalo rat liver line, BRL, promoted the growth and survival of the cells in culture. Mitotically active cells were identified in cultures that had undergone extensive differentiation. The embryo cell cultures provide an in vitro system for investigations of biochemical parameters influencing zebrafish neuronal cell growth and differentiation. 相似文献
11.
Walter E. Madsen Michael J. Walker Elizabeth A. Shaughnessy John M. Brown Tapas K. Das Gupta 《In vitro cellular & developmental biology. Plant》1990,26(10):971-977
Summary A new tumor cell line has been established from a malignant pleural effusion in a 28-yr-old female patient with a primary
alveolar rhabdomyosarcoma of the left buttock. The in vitro and in vivo growth characteristics, morphologic features, abnormal
karyotype, and immunohistochemical staining pattern indicate that this cell line is comprised of primitive malignant mesenchymal
cells derived from a human rhabdomyosarcoma. Receptor studies done on tumors grown in male athymic mice revealed a single
class of high affinity saturable cytoplasmic estrogen receptor (Bmax 2.6 fm/mg cytosol protein, Kd 0.34 mM). Likewise, sucrose density gradient analysis demonstrated specific low-capacity, high-affinity estradiol binding predominately
in the 8S region. Cell growth in monolayer culture and on soft agar in the presence of estradiol was inhibited by pharmacologic
concentrations of estradiol in a dose-responsive manner compared with control. We describe a newly characterized malignant
mesenchymal cell line derived from an alveolar rhabdomyosarcoma that is inhibited by pharmacologic doses of estradiol in vitro.
These findings suggest further investigation into the mechanism(s) of this estrogen-induced inhibition in rhabdomyosarcomas. 相似文献
12.
ZF-L cells were derived from normal adult zebrafish liver, and have been growing in culture for more than 100 generations. The cells were derived in basal nutrient medium supplemented with fetal bovine serum (FBS), trout serum, trout embryo extract, bovine insulin and mouse epidermal growth factor. After 50 generations in culture, optimal growth of the cells was achieved in medium supplemented with FBS (5%) and trout serum (0.5%). ZF-L cells were hypodiploid (modal chromosome number= 46) and exhibited an epithelial morphology. ZF-L cell homogenates exhibited alanine and aspartate aminotransferase, glucose-6-phosphatase and alkaline phosphatase enzyme activities. The cells synthesized and released several proteins into the culture medium, including a 70 kDa protein recognized by anti-bovine serum albumin IgG.Abbreviations NF
-naphthoflavone
- EGF
epidermal growth factor
- EROD
7-ethoxyresorufinO-deethylase
- FBS
fetal bovine serum
- PBS
phosphate-buffered saline
- PMSF
phenylmethylsulfonyl fluoride
- TCDD
2,3,7,8-tetrachlorodibenzo-p-dioxin 相似文献
13.
Augusto Pessina Elisabetta Mineo Maria Grazia Neri Laura Gribaldo Robert Colombi Paolo Brambilla Gintaras Zaleskis 《Cytotechnology》1992,8(2):93-102
A new murine cell line designated as SR-4987 was established by treating a long-term bone marrow culture with the supernatant
from Y-1 cells which actively produce viral C-particles (MuLV). The line showed a fibrolbast-like morphology and its mesodermal
origin was confirmed by immunocytochemical staining. Flow cytometric analysis of DNA index evidenced a tetraploid number of
chromosomes whereas cell cycle analysis showed 34.8% of cells in S phase and 60.7% in G1.In vitro growth studies demonstrated a population doubling time of 14.7h, a good plating efficiency (52.3%) and a very poor agar clonogenic
capacity (0.6%). SR-4987 was tumorigenic only in syngeneic mice in which sarcomas were induced. The line produced M-CSF in
the culture supermatant whereas G-CSF, IL-3 and GM-CSF were not detected. Studies are in progress to assess the production
of other cytokines and to verify if same autocrine growth factor is involved in the control of SR-4987 proliferation. Our
line provides a further model of stromal cells for studying the interaction between hemopoietic progenitors and their micro-environment,
as well as to study factors produced by stromal cells acting as modulators of proliferation and differentiation of related
cell populations. 相似文献
14.
Establishment of a human fetal cardiac myocyte cell line 总被引:4,自引:0,他引:4
Yi-Chong Wang Nicolas Neckelmann Ann Mayne Ahvie Herskowitz Alagarsamy Srinivasan Kenneth W. Sell Aftab Ahmed-Ansari 《In vitro cellular & developmental biology. Animal》1991,27(1):63-74
Summary Human cardiac myocytes undergo degeneration, cytolysis, and necrosis in a number of clinical disease conditions such as myocarditis,
dilated cardiomyopathy, and during episodes of cardiac allograft rejection. The precise cellular, biochemical, and molecular
mechanisms that lead to such abnormalities in myocytes have been difficult to investigate because at present it is not possible
to obtain and maintain viable cell cultures of human adult cardiac myocytes in vitro. However, human fetal cardiac myocytes
are relatively easy to maintain and culture in vitro, but their limited availability and growth, variability from one preparation
to another, and varying degrees of contamination with endothelial and epithelial cell types have made it difficult to obtain
reliable data on the effect of cardiotropic viruses and cardiotoxic drugs on such myocytes. These thoughts prompted us to
attempt to derive a cell line of human cardiac origin. Highly enriched human fetal cardiac myocytes were transfected with
the plasmids pSV2Neo and pRSVTAg and gave rise to a cell line (W1) which has been maintained in culture for 1 yr. Morphologic
and phenotypic analyses of W1 cells by flow microfluorometry and immunoperoxidase techniques indicate that the W1 cell line
shares many properties of human fetal cardiac myocytes, but appears not to react with specific antibodies known to react with
markers unique to human endothelial, epithelial, skeletal muscle, and dendritic cells. These preliminary data suggest that
the W1 cells may provide a unique source of an established cell line that shares many properties ascribed to human cardiac
myocytes.
This study was supported by grant 1RO1-25566-03 from the National Institutes of Health, Bethesda, MD, to A. Ahmed-Ansari and
by a Grant-in-Aid from the American Heart Association, Georgia Affiliate, to Nicolas Neckelmann. 相似文献
15.
The heat shock 70 family of proteins is one of the most highly conserved among all species. The genes encoding these proteins have been cloned and sequenced from bacterial species to humans with a high degree of homology preserved throughout evolution. Here we describe the cloning and characterization of a cDNA encoding a 70 kd heat shock cognate (hsc70) gene from the zebrafish (Danio rerio). A high degree of conservation is observed among hsc70 genes of other species as shown by phylogenetic analysis. The characterization of a hsc70 gene in the zebrafish provides a marker for studying the role of a constitutively expressed member of the hsp70 family in an important developmental and evolutionary model system. 相似文献
16.
Allan M. Crawford 《In vitro cellular & developmental biology. Plant》1982,18(10):813-816
Summary The first continuous coleopteran cell line, designated DSIR-HA-1179, was derived from the scarab beetleHeteronychus arator. Most cells are spindle-shaped, 25 to 40 μm long and 15 to 20 μm wide, adhere strongly to plastic surfaces, and are highly resistant to trypsin and collagenase treatment. The population doubling time of 6 d, at 27° C, is slow compared with most dipteran and lepidopteran cell lines. The cells cross-react in comparative radioimmunoassays withH. arator larval tissue but not other insect cell lines and support the replication ofOryctes baculovirus and at least two nodaviruses, black beetle virus, and Flock House virus. 相似文献
17.
Isaka K Nishi H Nakada T Osakabe Y Hokamura M Serizawa H Ebihara Y Takayama M 《Human cell》2002,15(4):200-206
We present a new cell line, EJ established from an invasive endometrioid adenocarcinoma of the uterine corpus in a 56-year-old patient. The cells show rapid growth in culture with a doubling time of 16 h and high migration activity. Monolayer-cultured cells were polygonal in shape showing a tendency to pile up without contact inhibition. Subcutaneous transplantation of the EJ cells into nude mice formed solid tumors that were histologically diagnosed as adenocarcinoma, whereas no metastasis was observed. Cultured EJ cells produced tissue polypeptide antigen (IPA). Genetic and molecular analyses revealed high telomerase activity but not estrogen receptor alpha expression. Using the DNA sequencing technique, we have screened EJ cells for p53 mutation in exon 5 to 8 but no mutation of p53 was observed. This cell line appears to represent the development of a more malignant clone with divergent receptor function and growth behavior, and provides us with an interesting new tool for the study of tumorigenesis in the human endometrium. 相似文献
18.
19.
A bioenergetics model was developed from observed consumption, respiration and growth rates for zebrafish Danio rerio across a range (18–32° C) of water temperatures, and evaluated with a 50 day laboratory trial at 28° C. No significant bias in variable estimates was found during the validation trial; namely, predicted zebrafish mass generally agreed with observed mass. 相似文献
20.
The success of a robust vertebrate inflammatory response is in part because of the migratory potential of its haematopoietic components. Once these cells converge at an inflammatory site, they interact with each other as well as non‐immune tissues and infectious agents to help manage both the scale and the duration of any ensuing response. Exactly how these blood cells, that constitute the innate and adaptive immune systems, contribute to such immune responses remain largely unknown. Traditionally, assessing these contributions relied upon histological analysis of fixed tissue sections complemented with in vitro dynamic data. Although informative, translating results from these studies into the multicellular whole‐animal setting remain difficult. Recently, non‐invasive live imaging of the immune system in animal models is providing significant insights into how immune cells function within their intact natural environment. Although the majority of these studies have been conducted within mice, another vertebrate, the zebrafish Danio rerio is being recognized as an ideal platform for non‐invasive live imaging applications. The optical transparency, rapid development, genetic tractability and highly conserved innate and adaptive immune systems of this well‐established developmental model have been exploited in a number of recent studies evaluating the immunocompetence of fluorescently tagged blood cells. In addition, similar live imaging studies are helping to dissect the ontogeny of blood‐cell development by tracking various haematopoietic precursor cells to assess their contribution to different blood lineages. This review will examine some recent advances that have helped D. rerio emerge as a live imaging platform as well as its potential to offer valuable insights into the genetics behind diseases associated with immune cell dysfunction. 相似文献