Hybrid cells derived from mouse and man: artificial heterokaryons of mammalian cells from different species.
TL;DR: Hybrid Cells Derived from Mouse and Man : Artificial Heterokaryons of Mammalian Cells from Different Species is presented, which describes how cells from different species can be hybrids.
Abstract: Hybrid Cells Derived from Mouse and Man : Artificial Heterokaryons of Mammalian Cells from Different Species
Citations
More filters
TL;DR: The derivation of a number of tissue culture cell lines which secrete anti-sheep red blood cell (SRBC) antibodies is described here, made by fusion of a mouse myeloma and mouse spleen cells from an immunised donor.
Abstract: THE manufacture of predefined specific antibodies by means of permanent tissue culture cell lines is of general interest. There are at present a considerable number of permanent cultures of myeloma cells1,2 and screening procedures have been used to reveal antibody activity in some of them. This, however, is not a satisfactory source of monoclonal antibodies of predefined specificity. We describe here the derivation of a number of tissue culture cell lines which secrete anti-sheep red blood cell (SRBC) antibodies. The cell lines are made by fusion of a mouse myeloma and mouse spleen cells from an immunised donor. To understand the expression and interactions of the Ig chains from the parental lines, fusion experiments between two known mouse myeloma lines were carried out.
19,053 citations
TL;DR: It appears that the cell surface of heterokaryons is not a rigid structure, but is ‘fluid’ enough to allow free ‘diffusion’ of surface antigens resulting in their intermingling within minutes after the initiation of fusion.
Abstract: Cells from established tissue culture lines of mouse ( cIID ) and human ( VA-2 ) origin were fused together with Sendai virus, producing heterokaryons bearing both mouse and human surface antigens which were then followed by the indirect fluorescent antibody method. Within 40 mm following fusion, total mixing of both parental antigens occurred in over 90% of the heterokaryons. Mouse H-2 (histocompatibility) and human surface antigens were visualized by successive treatment of the heterokaryons with a mixture of mouse alloantiserum and rabbit anti- VA-2 antiserum, followed by a mixture of fluorescein-labelled goat anti-mouse IgG and tetramethyl-rhodamine-labelled goat anti-rabbit IgG(Fc). The cIID x VA-2 fusions were carried Out in suspension and maintained at 37°C in a shaking water bath; aliquots were removed at various intervals and stained with the above reagents. The heterokaryon population was observed to change from an initial one (5-min post-fusion) of non-mosaics (unmixed cell surfaces of red and green fluorescence) to one of over 90% mosaics (total intermixing of the 2 fluorochromes) by 40 min after fusion. Mouse-human hybrid lines, derived from similar fusions, gave fluorescence patterns identical to those of the mosaic heterokaryons. Four possible mechanisms would yield such results: (i) a very rapid metabolic turnover of the antigens; (ii) integration of units into the membrane from a cytoplasmic precursor pool; (iii) movement, or ‘diffusion’of antigen in the plane of the membrane; or (iv) movement of existing antigen from one membrane site into the cytoplasm and its emergence at a new position on the membrane. In an effort to distinguish among these possibilities, the following inhibitor treatments were carried out: (1) both short- and long-term (6-h pre-treatment) inhibition of protein synthesis by puromycin, cycloheximide, and chloramphenicol; (2) short-term inhibition of ATP formation by dinitrophenol (DNP) and NaF; (3) short- and long-term inhibition of glutamine dependent pathways with the glutamine analogue 6-diazo-5-oxonorleucine; and (4) general metabolic suppression by lowered temperature. The only treatment found effective in preventing the mosaicism was lowered temperature, from which resulted a sigmoidal curve for per cent mosaics versus incubation temperature. These results would be consistent with mechanisms iii and/or iv but appear to rule out i and ii. From the speed with which the antigen markers can be seen to propagate across the cell membrane, and from the fact that the treatment of parent cells with a variety of metabolic inhibitors does not inhibit antigen spreading, it appears that the cell surface of heterokaryons is not a rigid structure, but is ‘fluid’ enough to allow free ‘diffusion’ of surface antigens resulting in their intermingling within minutes after the initiation of fusion.
758 citations
TL;DR: It is concluded that a novel membrane protein determines susceptibility to ecotropic MuLV infection by binding and/or fusion with the virus envelope.
683 citations
TL;DR: Malignancy can be suppressed when malignant cells are fused with certain non-malignant ones and the hybrid cells derived from such fusions give rise to segregants in which a loss of chromosomes is associated with reversion to malignancy.
Abstract: Malignancy can be suppressed when malignant cells are fused with certain non-malignant ones. The hybrid cells derived from such fusions give rise to segregants in which a loss of chromosomes is associated with reversion to malignancy. The expression of histo-compatibility antigens can also be suppressed when cells bearing these antigens are fused with others that express them poorly.
581 citations
TL;DR: During development and tissue repair the fusion of genetic and cytoplasmic material between cells of different origins is an important physiological process that could be important in the development of the cancer stem cell.
Abstract: Most tumours are derived from a single cell that is transformed into a cancer-initiating cell (cancer stem cell) that has the capacity to proliferate and form tumours in vivo. However, the origin of the cancer stem cell remains elusive. Interestingly, during development and tissue repair the fusion of genetic and cytoplasmic material between cells of different origins is an important physiological process. Such cell fusion and horizontal gene-transfer events have also been linked to several fundamental features of cancer and could be important in the development of the cancer stem cell.
529 citations
References
More filters
TL;DR: In this article, the optimal concentrations of amino acids, vitamins, nucleic acid constituents and various accessory growth factors for the maintenance of cell life in vitro have been established and a completely synthetic feeding solution has been devised.
Abstract: SummaryThe nutrition of animal cells in tissue culture has been studied with particular reference to amino acids, vitamins, nucleic acid constituents and various accessory growth factors. The optimal concentrations of these substances for the maintenance of cell life in vitro have been established and a completely synthetic feeding solution has been devised. Although this mixture will not support cell life indefinitely, it has been found adequate to maintain chick embryo cells for an average period of 4 to 5 weeks. The mixture is being used as a basal synthetic medium for further studies on the unidentified growth-promoting substances known to occur in natural media.
970 citations
TL;DR: The method described was developed to facilitate the analysis of chromosome complements in cells freshly isolated from monkey kidney cortex and growing on glass, and in “altered” monkey cells grown on glass or in suspension.
Abstract: The method described was developed to facilitate the analysis of chromosome complements in cells freshly isolated from monkey kidney cortex and grown on glass, and in “altered” monkey cells grown on glass or in suspension. Cells were treated with hypotonic solution (quarter-strength Tyrode or diluted medium) for 30 min, or with colchicine in a final concentration of 25 μg/ml (.0025%) for 12-18 hr followed by hypotonic salt solution for 5 min, then fixed in acetic alcohol (1:3) for 5 min. With cells centrifuged from suspended cultures, addition of fixative had to be gradual. Directly after fixation, films of cells on slides were air dried completely. This produces a more uniform and complete flattening of cells than can be achieved by manual pressure; yet, fragmentation of chromosome complements does not occur. Fixed and air dried slides may be stored for days without deterioration or they may be stained immediately in 2% natural orcein (G. T. Gurr, London) in 50% acetic acid. Preparations can be made perm...
468 citations
TL;DR: Two characteristic regions were observed in the reacting cells; one may be a part of the enfolded cell surface, and the other is a complexed projection on the cell surface at the contact point of opposed cells.
244 citations
191 citations
143 citations