agar and Leukemia

Current concepts of immunotherapeutic approaches in leukemias and lymphomas using activated cytotoxic lymphocytes and macrophages are briefly reviewed. Defective cellular immunocytotoxic activities and effects of interleukins and chemotherapeutic drugs thereupon are discussed. In vitro assays to measure lymphokine-activated killer (LAK) and natural killer (NK) cell activities suffer from various problems, depending on the quality of the endpoints. Our clonogenic microassay for LAK cell activity, using agar-containing glass capillaries, avoids some of the potential artifacts and offers several advantages that are discussed. As an example the stimulatory effect of low mafosfamide concentrations on the LAK cell activity versus K562 human myeloid leukemia cells is demonstrated. Thus, our clonogenic LAK microassay provides a valid tool for preclinical screening of immunomodulatory agents.

Topics: Agar; Cyclophosphamide; Cytotoxicity Tests, Immunologic; Cytotoxicity, Immunologic; Humans; Immunotherapy, Adoptive; Interferons; Interleukins; Killer Cells, Lymphokine-Activated; Killer Cells, Natural; Leukemia; Lymphoma; Tumor Cells, Cultured; Tumor Stem Cell Assay

Human T-T cell hybrids are developed by fusing activated T lymphocytes exhibiting a desired immunological function or producing soluble factors with a human tumor T cell line with the objective to immortalize the T cell properties of interest. Mutagenized human tumor T cell lines, deficient for the enzyme hypoxanthine-guanine phosphoribosyl transferase have been used for the development of T-T cell hybrids. Unfused tumor cells are removed by using appropriate selection media. Certain of these media contain components (such as thymidine) that inhibit the growth of the hybrids. A different method involves the use of tumor T cell lines chemically treated, before the fusion, with irreversible biochemical inhibitors. This treatment eliminated any unfused cells of the T cell line. Recently, a method has been developed for the generation of human T-T cell hybrids without the use of mutagenized or chemically treated tumor T cell lines. Hybrids are selected on the basis of their ability to form colonies in soft agar, and their hybrid nature is confirmed by HLA typing and functional tests. The human lymphoblastoid cell lines used did not form colonies in agar. Hybrids developed by this method exhibit excellent growth characteristics and increased stability. A large number of human T-T cell hybrids producing growth, differentiation or immunoregulatory factors have been developed. Certain hybrids exhibiting immunological functions requiring direct cell-cell contact have been developed also. The advantages of using T-T cell hybrids over other methods for immortalizing T cell functions or lymphokine production are summarized. Also, the obstacles in developing T-T cell hybrids are discussed.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Agar; Cell Fusion; Cell Line, Transformed; Clone Cells; Culture Media; Humans; Hybridomas; Hypoxanthine Phosphoribosyltransferase; Leukemia; Lymphokines; Selection, Genetic; T-Lymphocytes; Tumor Cells, Cultured

The technique of bone marrow cultures has been shown to be of value in childhood acute leukemia. It now appears that acute myelogenous leukemia may be due to defective maturation of normal progenitor cells. The pattern of growth of these cells has been demonstrated to be of prognostic value. In contrast, the growth of normal progenitor cells from the bone marrow cultures of children with acute lymphocytic leukemia (ALL) may be due to the few remaining normal cells. The cause of granulocytopenia in childhood ALL is still unclear.

Topics: Acute Disease; Agar; Bone Marrow; Cell Transformation, Neoplastic; Cells, Cultured; Child; Child, Preschool; Colony-Stimulating Factors; Culture Media; Granulocytes; Hematopoiesis; Humans; Leukemia; Leukemia, Lymphoid; Leukemia, Myeloid, Acute; Phytohemagglutinins

Topics: Agar; Anemia, Aplastic; Cell Count; Cell Differentiation; Clone Cells; Colony-Stimulating Factors; Culture Techniques; Hematopoiesis; Hematopoietic Stem Cells; Humans; Leukemia; Neutropenia

Topics: Agar; Animals; Blood Cells; Bone Marrow Cells; Cell Division; Cells, Cultured; Clone Cells; Cricetinae; Culture Media; Culture Techniques; Dogs; Guinea Pigs; Haplorhini; Hematopoietic Stem Cells; Humans; Leukemia; Leukocytes; Macrophages; Methods; Mice; Rabbits; Rats; Spleen

Topics: Agar; Animals; Child; Culture Media; Culture Techniques; Hodgkin Disease; Humans; Inclusion Bodies, Viral; Leukemia; Lymphoma; Lymphoma, Large B-Cell, Diffuse; Lymphoma, Non-Hodgkin; Male; Mice; Microscopy, Electron; Mycoplasma; Neoplasms; Neoplasms, Experimental; Rabbits; Rats

To determine the antiproliferative activity of Rubus parvifolius L. (RP) extract, its medicinal serum and RP total saponins (RPTS) against K562 cells in vitro and in vivo.. Nude mice models bearing leukemia tumors were treated with different concentrations of RP extract. The size, weight and histopathological change of leukemic tumors were determined. Semi-solid agar culture and methylthiazolyl tetrazolium (MTT) assay were used to determine in vitro the inhibition of colony formation and proliferation of K562 cells respectively by different concentrations of RP medicinal serum and RPTS.. RP extract had a tumor inhibition rate of 84.8% when administered to mice at a dose of 1.0 g/day of crude RP root equivalent. Semi-solid agar culture of K562 cells in the presence of 20% (v/v) of RP medicinal serum and 150 mg/L RPTS demonstrated a 50.8% and 100% inhibition of the colony forming unit (CFU)-K562, respectively. The same doses of RP medicinal serum and RPTS showed a proliferation inhibition of 31.4% and 86.3%, respectively against K562 cells in MTT assay.. RP extract and RPTS show effective antiproliferative activity against myeloid leukemia cells in vitro and in vivo.

Topics: Agar; Animals; Cell Proliferation; Chromatography, High Pressure Liquid; Humans; K562 Cells; Leukemia; Mice; Mice, Inbred BALB C; Mice, Nude; Plant Extracts; Rosaceae; Saponins; Subcutaneous Tissue; Tumor Stem Cell Assay; Xenograft Model Antitumor Assays

The peroxisome proliferator-activated receptor gamma (PPAR gamma) is a member of the nuclear receptor family that forms heterodimers with retinoid X receptor. These heterodimers bind to DNA and activate the transcription of target genes. Here, we report that the PPAR gamma receptor protein is expressed in primary myeloid and lymphoid leukemias and in lymphoma and myeloma cell lines. In this study, we compared the activity of several PPAR gamma ligands including BRL49653 (rosiglitazone), 15-deoxy-Delta 12,14-prostaglandin J(2), and the novel triterpenoid 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid on leukemia cells. Exposure to these PPAR gamma ligands induced apoptosis in myeloid (U937 and HL-60) and lymphoid (Su-DHL, Sup-M2, Ramos, Raji, Hodgkin's cell lines, and primary chronic lymphocytic leukemia) cells. A similar exposure to these PPAR gamma ligands induced the differentiation of myeloid leukemic cells. A combination of PPAR gamma ligands with a retinoid X receptor agonist (i.e., LG100268) or a retinoic acid receptor agonist (i.e., all trans-retinoic acid) enhanced differentiating and growth-inhibitory effects. 2-Cyano-3,12-dioxooleana-1,9-dien-28-oic acid induced differentiation and apoptosis with much greater potency than the other PPAR gamma ligands in established cell lines and primary chronic lymphocytic leukemia samples. Exposure to 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid induced mitochondrial depolarization and caspase activation, which was associated with apoptosis induction. In Bcl-2-overexpressing chronic lymphocytic leukemia cells, the small-molecule Bcl-2 inhibitor HA14-1 sensitized these cells to 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid-induced apoptosis. These results suggest that PPAR gamma ligation alone and in combination with retinoids holds promise as novel therapy for leukemias by activating the transcriptional activity of target genes that control apoptosis and differentiation in leukemias.

Topics: Agar; Apoptosis; Blotting, Western; Cell Differentiation; Cell Line; Cell Line, Tumor; Cell Nucleus; Cell Proliferation; Cell Survival; Dimerization; Fibrinolytic Agents; Flow Cytometry; HL-60 Cells; Humans; Imidazoles; Immunologic Factors; Jurkat Cells; Leukemia; Ligands; Oleanolic Acid; Phagocytosis; Plasmids; PPAR gamma; Prostaglandin D2; Proto-Oncogene Proteins c-bcl-2; Receptors, Retinoic Acid; Retinoic Acid Receptor alpha; Retinoid X Receptors; RNA, Messenger; Rosiglitazone; Thiazolidinediones; Transcription, Genetic; Transcriptional Activation; Transfection; U937 Cells

Peripheral blood cells from a female infant with Down syndrome and over 60% circulating myeloblasts were cultured in soft agar. Growth was virtually restricted to cluster formation, and cluster-forming cells resided almost exclusively in the very light density fraction (SG less than 1.062). Morphological assessment of clusters revealed no evidence of cellular differentiation beyond the blast cell stage. Despite receiving no specific chemotherapy, the peripheral blood normalized within 2 months, and there was no evidence of leukemia when the patient died aged 1 year from cardiac pathology. The findings indicate that caution should be exercised when assessing prognosis on the basis of in vitro growth characteristics in such patients.

Topics: Agar; Blood Cells; Cell Differentiation; Cells, Cultured; Down Syndrome; Female; Humans; Infant, Newborn; Leukemia; Myeloproliferative Disorders

The granulocyte-macrophage colony stimulating effect of bone marrow cells is shown to be less intensive than the stimulating effect of feeding leukocytes. It is supposed that these two stimulating substances activate different progenitor populations. Stimulating effect of mixture consisting of leukocytes and bone marrow cells are very close to that of bone marrow cells alone. Leukemic bone marrow cells demonstrate the least colony stimulating ability.

Topics: Agar; Cell Differentiation; Cells, Cultured; Colony-Forming Units Assay; Culture Media; Granulocytes; Hematopoietic Stem Cells; Humans; Leukemia; Leukocytes; Macrophages; Stem Cells

Studied were a total of 70 samples of blood and corresponding lactosera taken from lactating cows that were positive for leukosis on three farms with records for the disease. The lactosera were obtained from the milk samples through treatment in two ways--with calcium chloride and with chymosin. The experiments were carried out with untreated blood sera and with the lactosera obtained in both ways. Parallel to these, blood sera and lactosera were both studied in a concentrated state after being deeply frozen and centrifugated. The agar gel precipitation test (AGPT) was employed. It was concluded that the procedure described was a readily applicable method for laboratory use in the way of establishing enzootic leukosis, making unnecessary the blood sampling on a mass scale as undesirable from an economic and epizootic viewpoint.

Topics: Agar; Animals; Cattle; Cattle Diseases; Female; Gels; Immune Sera; Lactation; Leukemia; Milk; Precipitin Tests; Pregnancy

A transforming growth factor was found in the extracts of leukemic cells obtained from the peripheral blood of 11 patients with leukemia. This factor stimulated the colony formation of anchorage-dependent BALB/c 3T3 cells in soft agar. The high levels of colony-stimulating activities were observed in the cell extracts from patients with chronic myelogenous leukemia in blastic crisis (CML BC), acute myelogenous leukemia and CML in chronic phase. The factor from a CML BC patient was heat- and acid-labile, relatively stable to dithiothreitol treatment and inactivated by pronase treatment. Molecular size of the factor seems more than 10,000 daltons.

Topics: Acids; Agar; Animals; Cell Extracts; Cell Line; Cell Separation; Dithiothreitol; Hot Temperature; Humans; Leukemia; Leukocytes; Mice; Molecular Weight; Peptides; Transforming Growth Factors

A comparison was made between the agar and methylcellulose culture systems with respect to their ability to support the clonal growth of leukemic cells obtained from patients with acute myeloblastic leukemia, acute lymphoblastic leukemia, and chronic myelogenous leukemia in blastic crisis. The number of clusters and/or colonies formed and the morphology of the cells within them varied from patient to patient. Nevertheless, no significant difference between the two culture systems within given leukemic specimens was found. No significant differences were noted among three different conditioned media used as sources of colony-stimulating activity. Most of the cells within clusters and colonies were identified to be immature members of granulocyte-macrophage series or to be indistinguishable from the preculture leukemic blast cells by morphological and cell surface marker studies. Cells from myeloid crisis in chronic myelogenous leukemia grew well in the cultures, but cells from lymphoid crisis did not proliferate.

Topics: Adult; Agar; Aged; Antigens, Surface; Cell Division; Cells, Cultured; Female; Growth Substances; Humans; Leukemia; Leukemia, Myeloid, Acute; Male; Methylcellulose; Middle Aged; Rosette Formation

Human leukaemic cell specimens were obtained from patients and directly plated into soft agar (t = 0) or cultured for 1 week in liquid phase and then plated in soft agar. Growth for 1 week in liquid phase allowed the clonal growth in agar of leukaemic specimens which were unable to clone at t = 0. Clonal growth after liquid culture consisted of the usual leukaemic type of cluster-colonies, growth of a new type of 'syncytial' cell colony or a mixture of colony types. In addition, marrow from a patient with acute lymphocytic leukaemia produced normal-appearing colonies after 1 week of growth in liquid phase. These studies suggest a similarity in the growth requirements of some leukaemic cells and normal CFUd cells.

Topics: Adult; Agar; Aged; Cell Division; Cells, Cultured; Clone Cells; Culture Media; Female; Humans; Kinetics; Leukemia; Leukemia, Erythroblastic, Acute; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Leukocytes; Macrophages; Male; Middle Aged

Topics: Adult; Agar; Cell Division; Cells, Cultured; Colony-Stimulating Factors; Cytological Techniques; Hematopoiesis; Hematopoietic Stem Cells; Humans; Leukemia; Leukemia, Myeloid

Amethopterin synchronization of bone marrow and lymph node cells makes it possible to obtain well banded and elongated metaphases and prometaphases in the majority of patients with leukemia and lymphoma. Chromosomal analysis of most neoplasias is also possible through the use of mild tumor cell disaggregation methods, short term culture on feeder layers, and special media that help in the preferential proliferation of cancer cells. In the past, only about half of all patients with acute leukemia and lymphoma could be shown to have a chromosomal defect, and only a small proportion of solid tumors could be analyzed. With the new technology, abnormal tissue from the majority of cancer patients can be successfully studied and chromosomal abnormalities detected.

Topics: Adolescent; Adult; Agar; Bone Marrow; Bone Marrow Cells; Cell Line; Chromosome Aberrations; Chromosome Banding; Female; Humans; Leukemia; Lymph Nodes; Lymphocytes; Male; Metaphase; Methotrexate; Neoplasms; Tissue Extracts

This study was undertaken with the aim of identifying the different cell types found in human bone marrow by examining their surface morphology. In an attempt to obtain a homogeneous cell population, cells were both fractionated by discontinuous albumin density gradient centrifugation (DADGC) and selectively grown in nutrient agar. Both cell preparations underwent the critical point drying technique before examination under both the scanning electron microscope (SEM) and subsequently the light microscope (LM). When the SEM image of individual cells was compared with the corresponding LM image, it was not easy to identify the different cell types, because of the shrinkage and distortion that occurred during their preparation. The shrinkage observed under the SEM amounted to a 45% reduction in mean cell diameter. This shrinkage was confirmed by comparing the SEM and LM images of the same cell. Although shrinkage occurred throughout the dehydration sequence, critical point drying was responsible for a 25% reduction in mean cell diameter. Furthermore, direct observation under LM of fixed cells drying in air from ethanol, revealed visible contraction of the cell and distortion of the cell membrane. We assume that a similar morphological change occurred during critical point drying. We conclude that the shrinkage and distortion, caused by the dehydration process involved in SEM preparation, severely limit the value of a study of surface morphology by SEM in the identification of the different cell types found in human bone marrow.

Topics: Agar; Albumins; Bone Marrow Cells; Cell Fractionation; Cell Membrane; Cell Nucleus; Cell Separation; Cells, Cultured; Centrifugation, Density Gradient; Desiccation; Humans; Leukemia; Lymphocytes; Megaloblasts; Microscopy; Microscopy, Electron, Scanning; Temperature

Experiments were carried out to use the agar gel diffusion test (AGDT) in the study of bovine leukosis. An antigen was obtained from FLK-24 cells with a latent leukosis virus infection, which proved specific and suitable for AGDT. A total of 134 serum samples were examined applyig AGDT, for the presence of specific antibodies, originating from cattle of herds in which leukosis had been hematologically demonstrated. Investigated were also 56 sera of animals presenting an unknown clinical picture. Positive reaction was established in 70 per cent of the sera taken from herds with hematologically demonstrated leukosis. The specificity of AGDT was confirmed through establishing specific antibodies also in sera of lambs experimentally infected with a virus from a FBL-J 5 cell line with a latent infection. It is admitted that AGDT is a specific, sensitive, and readily applicable method, which can successfully be used in the diagnostics of leukosis in this country.

Topics: Agar; Animals; Antibodies, Viral; Antibody Specificity; Cattle; Cattle Diseases; Immunodiffusion; Leukemia; Leukemia Virus, Bovine; Serologic Tests

Topics: Agar; Animals; Cattle; Cattle Diseases; Immunodiffusion; Leukemia; Leukemia Virus, Bovine; Mass Screening

Hematopoietic cells from the blood or bone marrow (of leukemic and nonleukemic patients) grown in vitro using soft agar tissue-culture technics may be fixed in formalin, embedded in paraffin, sectioned, and mounted on glass slides. Light microscopic examination of these sections stained with hematoxylin and eosin and with other histologic stains provides information useful in investigative and diagnostic hematology. Morphologic interpretation of the characteristics of cultured cells is within the capability of pathologists and clinical hematologists. The slides provide a permanent record of growth in vitro.

Topics: Agar; Blood Cells; Bone Marrow Cells; Cells, Cultured; Culture Media; Formaldehyde; Histological Techniques; Humans; Leukemia; Paraffin

It was shown by the modified method of agar cultures that in the peripheral blood of a healthy man, aged from 4 days to 40 years. The number of cell precursors of granulocytes and macrophages (CFU-C) varied from 0.05 to 6.38 in adults and from 0.2 to 2.9 in children per 105 nuclear cells. CFU-C content in the patients with infectious mononucleosis and acute leukemia was 0.5--14 and 0--0.3 per 105 nuclear cells, respectively.

Topics: Acute Disease; Adolescent; Adult; Agar; Cell Division; Child; Child, Preschool; Clone Cells; Female; Gels; Humans; Infant; Infant, Newborn; Infectious Mononucleosis; Leukemia; Leukocytes; Male; Middle Aged

The influence of leukaemic cells from 12 patients with acute leukaemia on normal granulopoiesis in agar culture was investigated using leukeamic cell feeder layers. Leukaemic feeder cells from 7 of the 12 patients elicited no colony growth, while cells from the remaining 5 stimulated normal colony growth. In 3 of the 7 non-stimulatory patients release of inhibitory factors from the leukaemic cells seemed responsible for the effect on normal granulopoiesis, while inappropriate colony stimulating factor (CSF) production by the feeder cells could not be ruled out in the remaining 4 patients. When the leukaemic cells were cultured with, as well as without, conditioned medium, cells from 5 of the patients formed clusters. Growth in these cultures did not correlate to the effect found in the feeder layer experiments.

Topics: Acute Disease; Adolescent; Adult; Agar; Aged; Bone Marrow; Bone Marrow Cells; Cell Division; Cells, Cultured; Clone Cells; Colony-Stimulating Factors; Female; Granulocytes; Hematopoiesis; Hematopoietic Stem Cells; Humans; Leukemia; Leukocyte Count; Leukocytes; Male

Topics: Agar; Bone Marrow; Bone Marrow Cells; Cells, Cultured; Histocytochemistry; Humans; In Vitro Techniques; Leukemia; Leukemia, Myeloid, Acute; Lymphatic Diseases; Lymphocyte Activation; Plasma Cells

Topics: Adult; Agar; Bone Marrow; Bone Marrow Cells; Cell Division; Cells, Cultured; Child; Child, Preschool; Female; Hematopoiesis; Hematopoietic Stem Cells; Hematopoietic System; Humans; In Vitro Techniques; Infant; Leukemia; Leukemia, Lymphoid; Leukemia, Myeloid, Acute; Male; Middle Aged

Topics: Acute Disease; Agar; Cell Differentiation; Cell Division; Cells, Cultured; Cytological Techniques; Diffusion; Hematopoiesis; Hematopoietic Stem Cells; Humans; Leukemia; Micropore Filters

Topics: Aerobiosis; Agar; Cell Division; Culture Media; Leukemia; Microscopy, Electron; Mycoplasma; Serotyping

Topics: Agar; Animals; Blood; Bone Marrow; Bone Marrow Cells; Cells, Cultured; Culture Media; Culture Techniques; Dialysis; Female; Growth Substances; Hematopoiesis; Leukemia; Lymph Nodes; Male; Mice; Spleen; Thymus Gland; Urine

Topics: Agar; Animals; Antineoplastic Agents; Carbon Isotopes; Drug Resistance; Humans; In Vitro Techniques; Leukemia; Methods; Methylene Blue; Motion Pictures; Neoplasms; Neoplasms, Experimental; Neoplastic Cells, Circulating; Prognosis; Sulfhydryl Reagents; Tritium

Topics: Agar; Animals; Bone Marrow; Bone Marrow Cells; Cerebrospinal Fluid; Culture Techniques; Humans; Leukemia; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Mice; Retroviridae; Sepsis

Topics: Agar; Electrophoresis; Enzyme Activation; Fibrin; Fibrinolysin; Glycoproteins; Humans; Immune Sera; Immunoelectrophoresis; Leukemia; Leukocytes; Peptide Hydrolases; Protease Inhibitors; Trypsin Inhibitors

Topics: Agar; Animals; Cell Differentiation; Cell Transformation, Neoplastic; Clone Cells; Culture Techniques; Feedback; Hematopoietic System; Leukemia; Mice; Neoplasms, Experimental

Topics: Agar; Antineoplastic Agents; Female; Heparinoids; Humans; Leukemia; Male; Neoplasm Metastasis; Neoplasms; Radiation Effects

Topics: Agar; Alcoholism; Anemia, Hypochromic; Bone Marrow; Bone Marrow Cells; Cardiovascular Diseases; Culture Techniques; Fever; Gastrointestinal Diseases; Humans; Leukemia; Skin Diseases

Topics: Agar; Antimetabolites; Bacteriological Techniques; Blood Transfusion; Bone Marrow; Bone Marrow Diseases; Humans; Leukemia; Leukemia, Lymphoid; Mycoplasma; Penicillins; Radiotherapy

Topics: Agar; Animals; Antigens; Immunodiffusion; Leukemia; Leukemia, Experimental; Mice; Neoplasms; Oncogenic Viruses; Research

Topics: Agar; Antigens; Erythrocytes; Humans; Immune Sera; Leukemia; Spleen

Topics: Agar; Antigens; Erythrocytes; Humans; Immune System Phenomena; Leukemia