Showing papers on "Tissue culture published in 1969"
TL;DR: A cell line established in vitro from a spontaneous myeloid leukemia of SL strain mice was cytologically identified as myeloblast, and its normal differentiation appeared to be completely blocked in mass culture.
Abstract: A cell line was established in vitro from a spontaneous myeloid leukemia of SL strain mice. This cell line was cytologically identified as myeloblast, and its normal differentiation appeared to be completely blocked in mass culture. When the line cells were seeded in soft agar with a conditioned medium from normal cells, either macrophages or neutrophil granulocytes appeared from a single clone. The rate of formation of colonies containing differentiated cells always increased with an increase in the concentration of conditioned medium. The conditioned medium from this line cell was not as effective as was that from normal cells in inducing differentiation.
482 citations
TL;DR: A striking morphological change was observed in the cells adapted to culture growth; they appeared as mature neurons, while the cells of the tumor appeared as immature neuroblasts.
Abstract: Clonal lines of neurons were obtained in culture from a mouse neuroblastoma. The neuroblastoma cells were adapted to culture growth by the animal-culture alternate passage technique and cloned after single-cell plating. The clonal lines retained the ability to form tumors when injected back into mice. A striking morphological change was observed in the cells adapted to culture growth; they appeared as mature neurons, while the cells of the tumor appeared as immature neuroblasts.
Acetylcholinesterase and the enzymes for the synthesis of neurotransmitters, cholineacetylase and tyrosine hydroxylase were assayed in the tumor and compared with brain levels; tyrosine hydroxylase was found to be particularly high, as described previously in human neuroblastomas. The three enzymes were found in the clonal cultures at levels comparable to those found in the tumors. Similarly, there were no remarkable differences between the three clones examined.
461 citations
TL;DR: Mouse bone marrow cells in suspension were separated into a number of fractions on the basis of cell density by equilibrium density gradient centrifugation, or on the based of cell size by velocity sedimentation, to demonstrate that cells in some fractions formed more colonies in vivo than in the culture system.
Abstract: Mouse bone marrow cells in suspension were separated into a number of fractions on the basis of cell density by equilibrium density gradient centrifugation, or on the basis of cell size by velocity sedimentation. After each type of separation, the cells from the various fractions were assayed for their ability to form macroscopic spleen colonies in irradiated recipient mice, and for their ability to form colonies in a cell culture system. The results from either separation technique demonstrate that cells in some fractions formed more colonies in vivo than in the culture system, while cells in other fractions formed more colonies in culture than in the spleen. The results of control experiments indicate that this separation of the two types of colony-forming cells was not an artifact of the separation procedures. From these experiments it was concluded that the population of cells which form colonies in culture under the conditions used is not identical to the population of cells detected by the spleen colony assay.
405 citations
TL;DR: Mouse tumor C1300 has been established in tissue culture and cell attachment can induce morphological differentiation from an anaplastic round cell to a cell which has many properties of a mature neuron.
Abstract: Mouse tumor C1300 has been established in tissue culture. The cells have a round cell morphology in both the subcutaneous tumor and in suspension culture. However, when given a surface on which to attach, they send out processes up to 3 mm in length and assume the morphology of mature neurons. The attached cells are stained by the Bodian silver procedure for neurons, whereas the cells grown in suspension are not. Electron microscopy reveals that the attached cells contain neurofilaments, neurotubules, and densecore vesicles indicative of nerve fibers. Both free-floating and attached cells have tyrosine hydroxylase activity characteristic of sympathetic nervous tissue. Apparently cell attachment can induce morphological differentiation from an anaplastic round cell to a cell which has many properties of a mature neuron.
300 citations
TL;DR: Tissue culture cells of a Rous virus-induced rat tumor undergo syncytium formation when placed in contact with mouse embryo cell cultures infected with murine leukemia viruses, which can be used as a cytopathic end point for isolating and titrating these viruses in tissue culture.
Abstract: Tissue culture cells of a Rous virus-induced rat tumor undergo syncytium formation when placed in contact with mouse embryo cell cultures infected with murine leukemia viruses. This phenomenon can be used as a cytopathic end point for isolating and titrating these viruses in tissue culture. The principle should be applicable to detection of leukemia viruses of other species.
267 citations
240 citations
TL;DR: The evidence presented here demonstrates that the murine leukemia virus genome must be present in the original embryo cultures, and the possibility that the genetic information for making murines leukemia virus is present in a repressed form in every mouse embryo cell is discussed.
Abstract: BALB/c mouse embryo cells maintained in tissue culture on a schedule of rapid transfer at high cell density develop into tumorigenic lines that have lost contact inhibition of cell division. After several months in culture, certain of these lines begin to release mouse leukemia virus. This virus has properties indistinguishable from those of virus found in adult BALB/c mice. The evidence presented here demonstrates that the murine leukemia virus genome must be present in the original embryo cultures. The possibility that the genetic information for making murine leukemia virus is present in a repressed form in every mouse embryo cell is discussed.
155 citations
TL;DR: For example, the authors showed that normal and transformed hamster embryo fibroblasts, which communicate when cultured in medium containing fetal calf serum, appear to lose communication when cultured with medium containing calf serum.
Abstract: Epithelial cells of normal rat (adult) liver and hamster embryo in tissue culture communicate through membrane junctions: the membrane regions of cell contact are highly ion-permeable. Cancerous counterparts of these cells, cells from Morris' and Reuber's liver tumors and from x-ray-transformed embryo cultures, do not communicate under the same experimental conditions. These cells also fail to communicate with contiguous normal cells. Cancerous fibroblastic cells from a variety of tissues, including cells transformed by virus, x-radiation and chemicals, communicate as well as their normal counterparts; this is so for long- and short-term cell cultures. Communication in some fibroblastic cells is sensitive to components of blood serum: normal and transformed hamster embryo fibroblasts, which communicate when cultured in medium containing fetal calf serum, appear to lose communication in medium containing calf serum; the converse holds for hamster (adult) fibroblasts and 3T3 cells.
146 citations
TL;DR: In mouse cell lines transformed by SV40 virus, a decrease was observed in the higher ganglioside homologues disialo-ceramidetetrahexoside and monosialo, which indicates that the change is one of the virus-regulated functions, and it is postulated that it relates to the rejection of theirus-transformed cells.
Abstract: In mouse cell lines transformed by SV40 virus, a decrease was observed in the higher ganglioside homologues disialo-ceramidetetrahexoside and monosialo-ceramidetetrahexoside. Such change was observed only in cells which carry the virus genome, and it correlated with increased saturation density in tissue culture and with rejection in immunologically competent syngeneic host. This indicates that the change is one of the virus-regulated functions, and it is postulated that it relates to the rejection of the virus-transformed cells.
140 citations
TL;DR: Measles virus was propagated in monolayer cultures established from brain tissue of a patient with subacute sclerosing panencephalitis, and was identified by hemiagglutination-inhibition, but only in the supernatant of disrupted cultured cells.
Abstract: Measles virus was propagated in monolayer cultures established from brain tissue of a patient with subacute sclerosing panencephalitis. Syncytial cells were rendered fluorescent with measles specific antiserums only, by means of an indirect technique. The ultrastructural appearance of the microtubular aggregates was identical in brain tissue and in the cultured cells. Fusion experiments produced a cytopathic effect in humnan embryonic kidney and VERO cell cultures. The virus was identified by hemiagglutination-inhibition, but only in the supernatant of disrupted cultured cells.
TL;DR: In this article, the development of methods for routine cultivation of cells in vitro has provided the biochemist with a useful system for the study of cellular lipid metabolism, making it possible to obtain large quantities of a homogeneous population of cells.
Abstract: Publisher Summary The development of methods for routine cultivation of cells in vitro has provided the biochemist with a useful system for the study of cellular lipid metabolism, making it possible to obtain large quantities of a homogeneous population of cells. The degree of biological variation encountered in a tissue culture system is less than that obtained from studies employing whole animals. In addition, experimental conditions can be easily varied at will in cell culture systems. The differences between tissue culture systems and other experimental systems must be kept in mind when evaluating metabolic data obtained from in vitro cell studies. Most of the cells maintained in tissue culture are dedifferentiated in the sense that they do not carry out many of the specialized functions found in differentiated cells. These cells are not organized into specialized tissues as are cells in vivo, limiting some aspects of cellular interactions. Cells in culture are rapidly proliferating, and most lipid studies have been conducted with mixoploid cells derived from malignant tissue. The cellular growth environment of cells cultivated in vitro differs from that of cells in vivo.
Journal Article•
TL;DR: The addition of lymphocytes from patients suffering from recurrent aphthous ulceration, in either lymphocyte donor serum or inactivated foetal bovine serum, reduced significantly the level of survival of the oral epithelial cells as compared with the controls.
Abstract: In order to elucidate the possible role of humoral and cellular immune mechanisms in the pathogenesis of recurrent aphthous ulceration, a series of tissue culture experiments was undertaken in which oral epithelial cells were used as target cells. The survival of the oral epithelial cells was assessed, after culture for 24 hours, with the addition of sera, lymphocytes, or both sera and lymphocytes, from patients with recurrent aphthous ulceration and from persons free of the disease. In the experiments in which serum alone was added, there was no significant difference between the patients with recurrent aphthous ulceration and the controls.
The addition of lymphocytes from patients suffering from recurrent aphthous ulceration, in either lymphocyte donor serum or inactivated foetal bovine serum, reduced significantly the level of survival of the oral epithelial cells as compared with the controls.
TL;DR: The present studies suggest that complex carbohydrates are required for the adhesion of at least one type of animal cell and indicate that the conversion of nonadhesive to adhesive teratoma cells requires the synthesis of glycoproteins, glycolipids, and/or polysaccharides.
Abstract: Intercellular adhesion presumably involves components of the cell surface, but the chemical nature of these substances is not known. The present studies suggest that complex carbohydrates are required for the adhesion of at least one type of animal cell. Single cells obtained from “embryoid bodies,” the ascites-grown form of a mouse teratoma, aggregated in a complex tissue culture medium, but not in a glucose balanced salts solution. The active component of the tissue culture medium was identified as L-glutamine, and the only compounds found to replace it were the hexosamines D-glucosamine and D-mannosamine. A variety of studies indicated that the three compounds were active as a consequence of metabolic reactions. These results are consistent with known metabolic pathways and indicate that the conversion of nonadhesive to adhesive teratoma cells requires the synthesis of glycoproteins, glycolipids, and/or polysaccharides.
Journal Article•
TL;DR: This chapter discusses the role of physiological salt solutions used in fish tissue culture and presents a comprehensive source of information and a guide to the methods of fish cell and tissue culture.
Abstract: Publisher Summary This chapter presents a comprehensive source of information and a guide to the methods of fish cell and tissue culture. The evolution of fish cell and tissue culture follows a progression in time and complexity. Fish cell and tissue culture has been predominantly concerned with material from teleosts. Fish cells are comparable to mammalian cells in their response to freezing and storage at ultra-low temperatures, but like all poikilothermic animal cells, their rate of metabolism can be manipulated with temperature control. To a certain extent, the status of fish cell and tissue culture is also described in negative terms in the chapter. The chapter discusses the role of physiological salt solutions used in fish tissue culture.
TL;DR: The evidence suggests that the human antimalignant melanoma antibodies did not react with live malignant melanomas cells or with fixed or live cells from benign nevi or fixed guinea pig kidney, human kidney or HeLa cells.
Abstract: Cells were separated from tissue specimens of human malignant melanoma and grown in tissue culture on coverslips. After 8 days of growth the explanted cells were fixed and stored until used. Sera from 15 patients with malignant melanoma were reacted with the fixed melanoma cells and a variety of other cells. The indirect immunofluorescence method was used to test for presence of antibodies. Sera from all 15 melanoma patients reacted with fixed melanoma cells giving intense fluorescence of intracytoplasmic granules. The fluorescent material appeared to be independent of melanin granules. Homogenate of fresh melanoma but not of other neoplastic or normal tissue absorbed out the anti-melanoma antibodies. The human antimalignant melanoma antibodies did not react with live malignant melanoma cells or with fixed or live cells from benign nevi or fixed guinea pig kidney, human kidney or HeLa cells. The evidence suggests that these antibodies are unable to penetrate living cell membranes.
TL;DR: Collagen proline hydroxylase activity could be stimulated independently of cell growth by rapidly concentrating the cells indicating an activation process associated with cell density.
TL;DR: Physiologically different populations of cells appear to develop after priming and are able to respond in vitro in a macrophage-poor culture, some of the properties of these populations suggest that they are "memory cell" pools containing precursors of direct and indirect plaque-forming cells highly susceptible to a second antigenic stimulus.
Abstract: A cell suspension culture system combined with a procedure which separates most macrophages from lymphoid cells was used to investigate some of the cellular requirements for direct and indirect plaque-forming cell responses by nonprimed and primed mouse spleen cells in vitro. The plaque-forming cell response to heterologous erythrocytes in cultures of nonprimed spleen cells required both macrophages and lymphoid cells for its development. A significant indirect plaque-forming cell response did not develop in cultures of nonprimed spleen cells. In contrast, cultures of separated or macrophage-poor lymphoid cells from primed mice exhibited increasing responses relative to the response of unseparated spleen cells as the interval after priming increased. The cultures of separated lymphoid cells were not entirely free of phagocytic cells. Despite some evidence which suggests that these phagocytic cells had little function in the response, one cannot ascertain whether the lymphoid cells were responding directly to a second contact with antigen or whether the few contaminating phagocytic cells were performing a function essential to the response by the lymphoid cells. Physiologically different populations of cells appear to develop after priming and are able to respond in vitro in a macrophage-poor culture. Some of the properties of these populations suggest that they are "memory cell" pools containing precursors of direct and indirect plaque-forming cells highly susceptible to a second antigenic stimulus.
TL;DR: The phospholipid enhancement of cholesterol excretion was demonstrated in L5178Y cells prelabeling with exogenous cholesterol and L-cells prelabeled with biosynthesized cholesterol.
TL;DR: Glomeruli from normal and diseased human kidneys were explanted in tissue culture chambers and were studied by phase-contrast microscopy and time-lapse cinematography.
Abstract: Glomeruli from normal and diseased human kidneys were explanted in tissue culture chambers and were studied by phase-contrast microscopy and time-lapse cinematography. Observation from the time of exp
TL;DR: The conversion of simian virus 40 (SV40) component II deoxyribonucleic acid to component I has been used to assay polynucleotide ligase in extracts of tissue culture cells, finding increased activity was localized in the cytoplasm in vaccinia virus-infected cells.
Abstract: The conversion of simian virus 40 (SV40) component II deoxyribonucleic acid to component I has been used to assay polynucleotide ligase in extracts of tissue culture cells. All cell types examined, including chicken, hamster, mouse, monkey, and human cells, contained adenosine triphosphate-dependent ligase. After infection of mouse embryo, monkey kidney, and HeLa cells with polyoma virus, SV40, and vaccinia virus, respectively, the enzyme activity increased, but its cofactor requirement was unchanged. In vaccinia virus-infected cells, the increased activity was localized in the cytoplasm. Ligase induction occurred in the presence of cytosine arabinoside but was prevented by puromycin. Rifampicin blocked the production of infectious vaccinia particles but had little effect on the induction of ligase.
TL;DR: A clonally derived amelanotic melanoma cell line repeatedly has been forced to produce pigment by the inhibitor of DNA synthesis, I-β-D-arabinofuranosylcytosine (ara-C) at sublethal levels, and a decrease in rate of growth is found in other melanotic lines and is believed to be a significant factor in maintaining this differentiated function.
Abstract: A clonally derived amelanotic melanoma cell line repeatedly has been forced to produce pigment by the inhibitor of DNA synthesis, I-beta-D-arabinofuranosylcytosine (ara-C) at sublethal levels. One ara-C-derived melanotic line has been cloned, and has continued to produce pigment for 2 years on normal medium. The inhibitor is most effective when administered to synchronized cells in four pulses on successive days at 1.8 x 10(-5)M during the S phase of the cell cycle. Colcemid at a sublethal concentration, and growth on medium solidified with agar also evoked pigment production in this line, but a large number of other inhibitors of biosynthetic processes did not, under the conditions tested. The melanotic lines are active producers of tyrosinase (DOPA oxidase), whereas the amelanotic line produces an inhibitor of tyrosinase activity. Both enzyme and inhibitor are labile at 4 degrees C and -20 degrees C, and decay of the inhibitor in homogenates of amelanotic cells reveals a low level of residual DOPA oxidase activity. The mean population doubling time of a cloned melanotic line is 23 hr, and that of a cloned amelanotic line 16.5 hr. A similar decrease in rate of growth is found in other melanotic lines and is believed to be a significant factor in maintaining this differentiated function. Rapid growth may be related to the production of an inhibitor by the amelanotic cells.
TL;DR: A clonal line of astrocytes (designated CHB) derived from a brain tumor of human origin has been established in tissue culture and is capable of synthesizing an acid protein unique to the nervous system (S100-protein).
Abstract: A clonal line of astrocytes (designated CHB) derived from a brain tumor of human origin has been established in tissue culture. This cell line is capable of synthesizing an acid protein unique to the nervous system (S100-protein). The S100-protein synthesized by CHB cells is immunologically indistinguishable (by micro-complement fixation) from S100-protein present in human brain.
TL;DR: The tissue cultures of all cultivars studied have the capability to form roots regardless of the length of time in culture; root formation was enhanced by treatment with dalapon (2,2-dichloropropionic acid).
Abstract: Sugarcane tissues subcultured for over 4 years had lost the capability to differentiate shoots. In freshly isolated tissues from 3 cultivars, we readily obtained shoot differentiation on Murashige-Skoog medium, but shoot differentiation was lost, apparently irreversibly, after more than one subculture on a modified White's medium. Tissue from a fourth cultivar produced only roots. The tissue cultures of all cultivars studied have the capability to form roots regardless of the length of time in culture; root formation was enhanced by treatment with dalapon (2,2-dichloropropionic acid).
TL;DR: These findings, plus the demonstration that methylmalonyl-CoA mutase apoenzyme activity in whole cell homogenates from these mutant lines was not impaired, indicate that B 12 dependent methylmalonicaciduria is caused by a defect in the biosynthesis or metabolism of 5′-deoxyadenosylcobalamin.
TL;DR: N Nosema bombycis spores inoculated into cultures of Bombyx mori cells resulted in infection of the cells, and a form having two nuclei, similar to those of the sporoplasm, was seen among the newly formed spores.
TL;DR: It was confirmed in this study that tissue culture fluid harvested from rabies virus infected cell culture and inactivated by BPL or UV irradiation can be used for immunization against rabies.
Abstract: Summary and ConclusionsIt was confirmed in this study that tissue culture fluid harvested from rabies virus infected cell culture and inactivated by BPL or UV irradiation can be used for immunization against rabies. The protective activity of such preparation in mice was, however, generally not much higher than that of the NIH reference vaccine in a mouse potency test.