Abstract: A culture medium and method for the isolation and culture of plant protoplasts is disclosed which includes in it a quantity of neutral mammalian blood serum. An addition of 1 to 25% serum to protoplast media results in a greater yield of viable protoplast cells, helps to protect the protoplasts in culture, and makes it possible to obtain and maintain corn protoplasts, a heretofore difficult task.

Abstract: On the new findings that cyclodextrin shows no cytotoxicity or only slight cytotoxicity on cell growth and that lipophilic substances such as unsaturated fatty acids and lipophilic vitamins when present together with, or included in, cyclodextrin show such effects as cell growth promoting effect and accelerating effect of the productivity of valuable products, a serum-free or serum-reduced culture medium or a substitute composition for serum for a culture medium comprising a cyclodextrin and at least one lipophilic nutrient substance, said cyclodextrin and said lipophilic nutrient substance being preferably in the form of inclusion complex between them, is provided.

Abstract: A line of human blood cells which have high levels of oxidative activity (such as oxygenase, oxidase, peroxidase, and hydroxylase activity) is disclosed. Such cells grow in suspension culture, and are useful to determine the mutagenicity of xenobiotic substances that are metabolized into toxic or mutagenic substances. Mutation assays using these cells, and other cells with similar characteristics, are also disclosed.

Abstract: Antibodies and antigens are disclosed which provide a method of detecting and a method of combating flukes. A diagnostic method for the determination of an active fluke infection in a warm-blooded animal is provided which comprises the testing of body serum or body fluids for the presence of fluke spine glycoprotein or anti-spine antibodies. Fused cell hybrids ATCC HB-8086, ATCC HB-8087 and ATCC HB-8088, as well as the antibodies produced by the fused cell hybrids, are also provided.

Abstract: Human hybridomas producing a preselected human monoclonal antibody are prepared by fusing human lymphocytes with a hybrid fusion partner. The hybrid fusion partner is the result of fusing human lymphocytes with human myeloma cells at least once wherein the resulting hybrid cell is capable of being a functional human fusion partner.

Abstract: Cells synthesizing DNA are detected in a rapid, non-radioactive assay using the monoclonal antibody reagent secreted by the hybridoma produced in accordance with the present invention. The assay is used to study DNA repair in cells that have been exposed to various environmental toxins, chemotherapeutic agents, and the like. The monoclonal antibody secreted by the hybridoma is a valuable reagent for research and diagnosis.

Abstract: Monoclonal antibodies to theophylline having 5% or less cross-reactivity with caffeine and the continuous hybrid monoclonal cell lines for their production are provided. These antibodies are useful in a particle-enhanced turbidimetric inhibition immunoassay for theophylline.

Abstract: Monoclonal antibodies demonstrating a reactivity with human breast cancer are produced. The hybridoma cultures secreting immunoglobins are produced by hydridoma technology. Splenic lymphocytes of mice, immunized with membrane-enriched fractions of metastatic human mammary carcinoma tissue are fused with the NS-1 non-immunoglobulin-secreting murine myeloma cell line. Screening of immunoglobulin reactivities and double cloning of cultures yielded 11 monoclonal antibodies that demonstrated activities with the surface of human mammary tumor cells and not with the surface of apparently normal human tissues. These monoclonal antibodies aid in the diagnosis, prognosis and treatment of human breast cancer.

Abstract: Monoclonal antibodies to theophylline having 5% or less cross-reactivity with caffeine and the continuous hybrid monoclonal cell lines for their production are provided. These antibodies are useful in a particle-enhanced turbidimetric inhibition immunoassay for theophylline.

Abstract: This invention relates to a novel cell growth-promoting material for use as a supplement to basal tissue culture media for the cultivation of animal cells in vitro and to the method for its preparation. More particularly, it relates to an infusion consisting of growth-promoting material from adult, calf or fetal bovine blood clots as a supplement to basal media for cultivation of animal cells in vitro.

Abstract: This invention relates to the detection of antibodies in sera of AIDS and pre-AIDS patients and describes the biochemical and immunological analysis of antigens associated with the virus HTLV-III Human T-Cell Leukemia Virus. It is shown that antigens associated with the infection of human cells by this virus are specifically recognized by antibodies from AIDS patients. Specifically, HTLV-III isolated from AIDS patients and transmitted by cocultivation with an HT cell line is specifically detected by antibodies from human sera taken from AIDS patients. The method of detection of antibodies preferred is a strip radioimmunoassay (RIA) based on the Western Blot technique or an ELISA (an enzyme-linked immunosorbent assay) or an indirect immunofluorescence assay.

Abstract: A process for isolating and purifying a precipitated heterologous protein from a host cell culture, including treating the host cell culture with a buffered solution of ionic strength suitable to solubilize most of the host protein but in which refractile heterologous protein is substantially insoluble, and disrupting the cells to form a supernatant and an insoluble fraction; treating the insoluble fraction with a strongly denaturing solution to solubilize the refractile heterologous protein; and recovering renatured heterologous protein.

Abstract: Immortal, antibody-producing, hybridomally-produced clones which produce antibodies that react specifically with human HLA antigens and which are produced from rat lymphocyte cells sensitized against cells from a rat strain having different histocompatibility antigens are disclosed along with methods of producing the clones and antibodies and methods of using the antibodies in tissue crossing tests.

Abstract: Droplets having an electrical charge on the surface thereof are prepared from a first liquid and dispersed in a second liquid, a sterile aqueous tissue culture medium containing serum proteins. The first liquid is non-toxic to living cells and is relatively immiscible with water. The electrical charge on the surfaces of the droplets has a charge density in the range of from about 10 to about 100 charges per square nanometer. Serum protein films adhere to the charged droplets strongly enough to prevent rupture by human fibroblast cells grown thereon.

Abstract: A process for maintaining refractile proteins in solubilized form by replacing the strongly denaturing solution with a weakly denaturing solution.

Abstract: An assay that facilitates screening of hybridoma culture media for monoclonal anti-idiotype antibodies, particularly murine monoclonal antibodies that are useful for treating human B cell tumors is disclosed. The assay is a solid phase type assay and involves: incubating a lysate of the patienREFERENCE TO GOVERNMENT GRANT OR CONTRACTThe invention described herein was made in the course of work under a grant or contract from the National Institutes of Health.

Abstract: Hybridoma tumor cell line A.T.C.C. No. HB8116. An anti-H-Y antigen monoclonal antibody substance, "Hyclonalan," produced by said cell line. Use of Hyclonalan in immunoselection methodology.

Abstract: A process for purifying heterologous proteins from higher molecular weight components, including dissolving the heterologous protein in a strong denaturing solution and removing the higher molecular weight components using a molecular sieve or high speed centrifugation.

Abstract: A process for dissolving refractile proteins from their insoluble form by using a strongly denaturing solution.

Abstract: Antibody against human chorionic gonadotropin is raised in rats, preferably Lewis Strain Inbred rats, using either whole hormone or the .beta.-subunit as the antigen, and displays low cross-reactivity with luteinizing hormone as well as high sensitivity.

Abstract: An assay for HLA D phenotype wherein antigen-pulsed monocytes are contacted with antigen-specific T lymphocytes or T cell hybridomas and the extent of selective binding between the pulsed monocytes and the T lymphocytes or T cell hybridomas determines HLA D phenotype of the monocyte. The assay is particularly useful for quickly typing donor tissue prior to transplantation.

Abstract: A process for the recovery of immunoglobulins of high purity and potency from milk, in which process milk is passed through a re-usable immunoadsorbent column comprising an insoluble carrier material to which is bound a low-affinity monoclonal antibody specific to one or more milk immunoglobulins but not specific to any other common constituent of milk, the antibody bind immunoglobulin molecules are released by eluting the immunoadsorbent.

Abstract: Human interleukin 2 (IL-2) derived from induced human malignant cells has been purified to homogeneity using multiple high performance liquid chromatography (HPLC) steps. The purified IL-2 exhibits potent activity promoting the long-term in vitro culture of antigen-specific effector T-lymphocytes and in modulating lymphocyte reactivity.

Abstract: Monoclonal antibodies, and a cell line characterized by its production of such monoclonal antibodies, demonstrating specific reactivity to an antigenic determinant possessed by a plurality of types of adenoviruses, a method of isolating such cell lines, and the use of such antibodies for diagnostic and therapeutic purposes as well as for identifying chemical compounds with similar properties, are disclosed.

Abstract: A method for the extraction of intact recombinant human immune interferon with guanidine-HCl is disclosed. This method permits the purification to homogenity of intact recombinant human immune interferon.

Abstract: Antibodies having binding affinity for two desired antigens, hereinafter "recombinant monoclonal antibodies"; recombinant monoclonal antibodies produced by a quadroma cell or a trioma cell; and methods for producing recombinant monoclonal antibodies by means of a quadroma cell or a trioma cell, wherein a quadroma cell is the fusion product of a hybridoma cell which produces an antibody having specific binding affinity to one desired antigen and a hybridoma cell which produces an antibody having specific binding affinity for another desired antigen, and wherein a trioma cell is the fusion product of a hybridoma cell which produces an antibody having specific binding affinity to one desired antigen and a lymphocyte which produces an antibody having specific binding affinity to another desired antigen.

Abstract: A medium is provided which is useful in a fast and convenient system for multiplying cassava plants vegetatively by means of in vitro micropropagation of nodal cultures. This medium includes in parts per million the following:______________________________________ about 1200 NH.sub.4 NO.sub.3 from about 1000-1500 KNO.sub.3 about 300 MgSO.sub.4.7H.sub.2 O from about 150-300 NaH.sub.2 PO.sub.4.H.sub.2 O from about 150-300 CaCl.sub.2.2H.sub.2 O from about 18.5-37 Na.sub.2.EDTA from about 13.9-27.8 FeSO.sub.4.7H.sub.2 O. ______________________________________and suitable amounts of essential ingredients for a cassava micropropagation medium.In paticular, the medium also contains in part per million about: 3.0 H.sub.3 BO.sub.3, 10.0 MnSO.sub.4.H.sub.2 O, 6.0 ZnSO.sub.4.7H.sub.2 O, 0.25 Na.sub.2 MoO.sub.4.2H.sub.2 O, 0.025 CuSO.sub.4.5H.sub.2 O, 0.025 CaCl.sub.2.6H.sub.2 O, 0.75 KI, 2.5 Nicotinic acid, 10.0 Thiamine HCl, 1.0 Pyridoxine HCl, 100.0 M-inositol, 0.5 Glycine, 0.5 Folic acid, 0.05 Biotin, 0.

Abstract: A rat myeloma cell line which does not express an immunoglobulin chain, YB2/3.0.Ag.20, is prepared from the cell line Y3-Ag 1.2.3 via a hybrid myeloma cell line. This cell line, and variants thereof prepared by passaging and/or cloning the line, may be fused with immunocyte cells from an animal sensitized to an immunogen to produce hybrid myeloma cell lines which provide a source of monoclonal antibodies to said immunogen.

Abstract: Process for preparing a continuous and spontaneous human-interferon-producing lymphoblastoid cell line which involves (a) transforming human lymphocytes isolated from a human infant from birth up to about 2 years or an aborted human fetus, the transforming being carried out with Epstein Barr Virus and (b) selecting from the resulting transformed cell lines a cell line producing more than 1000 units of interferon per milliliter when cultivated at a cell density of 10.sup.6 cells per milliliter in the absence of any interferon inducer.

Abstract: A method of culture of human cells is disclosed which comprises effecting the cultivation in a culture medium containing an extract of micro algae, such as Chlorella, Scenedesmus or Spirulina, said method permitting the normal successive cultivation of human cells to be maintained efficiently without any morphological and genetic mutations over a greater number of successive of generations than has hitherto been possible even by the incorporation of animal serum in the culture medium, even when the addition amount of such animal serum is reduced substantially or animal serum is completely excluded.

Abstract: Monoclonal antibodies which are heavy-chain specific for porcine Ig's are secreted by novel hybrid cell lines produced by fusing SP2/O myeloma cells with B-lymphocytes from BALB/c mice immunized against porcine Ig's. The monoclonal antibodies are useful in porcine immunological research and pathological diagnosis.

Abstract: Disclosed herein is a process for producing erythropoietin, comprising bringing a material containing erythropoietin into contact with an adsorbent having a monoclonal anti-erythropoietin antibody, adsorbing erythropoietin on the adsorbent, and eluting the absorbed erythropoietin from the adsorbent.

Abstract: A non-transformed cell line is produced by treating BALB/c derived 10E2 cells with 5-bromodeoxyuridine and 5-iodo-2'-deoxyuridine to produce thymidine kinaseless cells which upon multiple cloning show a flat epitheloid appearance indicative of their non-transforming potential. These cells are used to determine the tumorigenic transforming potential of any gene by introducing the gene into the cells of the non-transformed cell line along with the Herpes simplex virus thymidine kinase gene which serves as a vehicle for cotransfection. The transformation of the cells is indicative of tumorigenic potential.

Abstract: A continuous hybridoma cell line which secretes recoverable quantities of monoclonal antibodies having specificity against Vitamin B.sub.6, which antibodies are useful in a method for detecting the presence of vitamin B.sub.6 in an animal sample.

Abstract: Novel method, cells and compositions are provided involving transforming B-lymphocytes to provide immortalization for continuous production of monoclonal antibodies to a predetermined ligand. T-cell free B-lymphocytes are combined with an Epstein-Barr virus transformed cell sensitive to a cytotoxic agent which does not significantly affect the B-lymphocytes under conditions where the sensitive EBV transformed cell acting as the transfer agent is killed and efficiently transforms the B-lymphocyte recipient cells with EBV. The EBV transformed B-lymphocyte cells are amplified and cloned, the desired clones isolated in accordance with conventional techniques and then used for production of monoclonal antibodies.

Abstract: Cultivation and reproduction of insect cells in a nutrient medium in which fetal calf serum is partially or completely replaced by egg yolk.

Abstract: Novel human lymphoblastoid cells and hybridomas derived therefrom are provided. The cells are a HGPRT negative human B-cell line. The cells are readily fusible with lymphoid cells to produce hybridomas which secrete human monoclonal antibodies.

Abstract: Human T-lymphoblast cell line, Proteinaceous products produced therefrom, messenger RNA and DNA expressing the proteinaceous products. A human T-lymphoblast cell line (Mo) maintained as a continuous culture constitutively produces proteins, including immune interferon, neutrophil migration inhibition factor, granulocyte-macrophage colony-stimulating activity and erythroid-potentiating activity, as well as other proteins produced by T-cells.

Abstract: Novel compositions for use as cell growth-promoting materials are made by the following novel process involving the steps of:(a) slowly contacting serum or plasma with sufficient chilled perchloric acid to reach a 0.1 to 0.25 final molar concentration of said perchloric acid in said serum or plasma,(b) at a temperature of -1.degree. C. to 15.degree. C.

Abstract: A human non-secretory plasmacytoid continuous cell line, established for five years in more than 150 passages, is karyotypically normal, easily grown and has the characteristic features of a plasmablast excepting for its secretory defect, and can be used for the preparation of human-human hybridomas with human B-lymphocytes and separation of the resulting hybridomas from the plasmacytoma cell line by growth in CO.sub.2 -containing media, or by fluorescence activated cell sorting, or both.

Abstract: Methods featuring, in one aspect, a method of preparing an antigen or mixture of antigens substantially free of antigens specific to at least two monoclonal antibodies, said method involving contacting an antigen mixture with one or more previously-isolated monoclonal antibodies to form complexes between those antibodies and antigens present in the mixture specific to the antibodies, removing the complexes from the antigen mixture to yield a partially purified antigen mixture, immunizing an animal with the partially purified antigen mixture, fusing spleen cells from the immunized animal to myeloma cells to form hybridomas capable of producing additional monoclonal antibodies, culturing said hybridomas to produce said additional monoclonal antibodies, contacting a sample of the partially purified antigen mixture with said additional monoclonal antibodies to form complexes between said additional monoclonal antibodies and antigens present in the antigen mixture specific to the additional monoclonal antibodies,

Abstract: Methods are disclosed for isolating and culturing human mammary epithelial cells of both normal and malignant origin. Tissue samples are digested with a mixture including the enzymes collagenase and hyaluronidase to produce clumps of cells substantially free from stroma and other undesired cellular material. Growing the clumps of cells in mass culture in an enriched medium containing particular growth factors allows for active cell proliferation and subculture. Clonal culture having plating efficiencies of up to 40% or greater may be obtained using individual cells derived from the mass culture by plating the cells on appropriate substrates in the enriched media. The clonal growth of cells so obtained is suitable for a quantitative assessment of the cytotoxicity of particular treatment. An exemplary assay for assessing the cytotoxicity of the drug adriamycin is presented.

Abstract: A process for producing anti-IL-2 antibody from hybridoma cells generated by fusing activated, IL-2 immunized, murine lymphocyte cells with neoplastic murine myeloma cells. Fusion is accomplished by mixing the two cell lines together in the presence of a fusing agent. After fusion, the hybridoma cells are cultured in vitro in a supplemented tissue culture medium to thereby produce anti-IL-2 antibody. Also, the hybridoma cells are cloned by a limiting dilution procedure to isolate even more potent sources of anti-IL-2 antibody. Anti-IL-2 antibody is then purified from either tissue culture medium conditioned by hybridoma cells, or from peritoneal ascites of mice challenged with hybridoma cells.

Abstract: A process for preparing murine IL-2 from malignant neoplastic cells includes culturing murine leukemia or lymphoma cells in vitro in a protein containing medium supplemented with various additives. An optimum concentration of a T cell mitogen is added to the culture medium to stimulate maximum production of a supernate which contains IL-2. After a period of time, the supernate is collected and purified into more concentrated form. Phorbol myristate acetate may be added to a suboptimum concentration of the T cell mitogen to reduce the quantity of the mitogen required to produce maximum quantities of murine IL-2.

Abstract: A process for constitutively producing murine IL-2 from hybridoma cells generated by fusing nitrogen stimulated malignant murine cells with drug sensitive murine thymoma driver cells. A fusing agent is used to fuse the two parent cells together. After fusion, the hybrid cells are cultured in vitro in a supplemented, serum containing tissue culture medium to thereby constitutively produce IL-2. The medium also includes a group of suppressing compounds which will prevent unfused driver cells from replicating, and feeder cells used to nurture the growth of competent hybrid cells.

Abstract: A process for producing murine IL-2 from malignant neoplastic cells which are incapable of IL-2 production by mitogen stimulation alone includes culturing murine leukemia or lymphoma cells in vitro in a protein-containing medium supplemented with various additives. A T cell mitogen and IL-1 are added to the culture medium as co-stimulants inducing the production of a supernate which contains IL-2. After a period of time, the supernate is collected and then assayed for IL-2 activity. Also in the present invention, IL-1 is utilized as a co-stimulant together with a suboptimum concentration of a T cell mitogen to induce IL-2 production in cell lines capable of generating IL-2 by mitogen stimulation alone. The use of IL-1 in these instances reduces the quantity of mitogen required to produce maximum levels of IL-2.

Abstract: A process for preparing IL-2 from human malignant cells includes culturing human leukemia or lymphoma cells in vitro in a serum containing medium supplemented with various additives. The culture is stimulated by an optimum concentration of a T cell mitogen to produce a supernate which contains IL-2. After a period of time, the supernate is collected and processed to purify the IL-2. Phorbol myristate acetate may be added to the culture medium to boost production of IL-2.

Abstract: The present invention relates to processes for inserting DNA into eucaryotic cells, particularly DNA which includes a gene or genes coding for desired proteinaceous materials for which no selective criteria exist. The insertion of such DNA molecules is accomplished by cotransforming eucaryotic cells with such DNA together with a second DNA which corresponds to a gene coding for a selectable marker.The invention further relates to processes for inserting into eucaryotic cells a multiplicity of DNA molecules including genes coding for desired proteinaceous materials by cotransformation with the desired genes and with amplifiable genes for a dominant selectable marker in the presence of successively higher amounts of an inhibitor.

Abstract: Human hepatoma cell lines, useful for metabolic studies such as screening potential carcinogens and mutagens, for cultivation of viruses, and for preparation of vaccines is obtained by culturing human hepatocarcinoma or hepatoblastoma on lethally irradiated cell feeder layers in the presence of a culture medium.

Abstract: A serum-free and mitogen-free T-cell growth factor (interleukin-2) preparation is prepared from human, bovine, or procine peripheral mononuclear blood cells which are washed several times with a liquid tissue culture medium and then stimulated in a tissue culture medium supplemented with serum and mitogen. The separated stimulated cells are again washed with fresh tissue culture medium to remove substantially all of the serum and mitogen. The washed cells are suspended in fresh tissue culture medium and conditioned under incubation conditions to transfer the growth factor into the liquid. The tissue culture medium separated from the stimulated cells can be recycled to stimulate additional cells. The supernatant can be concentrated from 50 to 100-fold on an ultrafilter. The supernatant can activate the production of natural killer cells in patients suffering from tumors.