Uridine uptake in a unicellular eukaryote during the interdivision period and after growth arrest

The rate of uridine uptake was measured in Tetrahymena after shiftdown to non-nutrient physiological salt solution. Uptake follows Michaelis-Menten kinetics and an apparent K_m of transport of 2 × 10⁻⁶ M has been estimated. This value is in good agreement with those reported for tissue-derived cells in culture. Incorporation of uridine into RNA follows similar kinetics suggesting that uptake is rate limiting for incorporation. Within three hours after shiftdown the rate of uptake is decreased by an order of magnitude. Also at three hours after shiftdown pairing occurs between cells of complementary mating types. It seems likely that the change in uptake is a reflection of a surface change associated with differentiation. The rate of uptake was also measured during the interdivision period using cells synchronized by a physical selection procedure. A change in rate occurs at the time the cells begin replication of DNA and is essentially stable thereafter. These results indicate that there exists in Tetrahymena a relationship between surface properties as assayed by uridine uptake and properties of growth and differentiation.     

Gompertzian growth pattern correlated with phenotypic organization of colon carcinoma,malignant glioma and non-small cell lung carcinoma cell lines

Abstract. In the current study we present a Gompertzian model for cell growth as a function of cell phenotype using six human tumour cell lines (A-549, NCI-H596, NCI-H520, HT-29, SW-620 and U-251). Monolayer cells in exponential growth at various densities were quantified over a week by sulforhodamine B staining assay to produce cell-growth curves. A Gompertz equation was fitted to experimental data to obtain, for each cell line, three empirical growth parameters (initial cell density, cell-growth rate and carrying capacity – the maximal cell density). A cell-shape parameter named deformation coefficient D (a morphological relationship among spreading and confluent cells) was established and compared by regression analysis with the relative growth rate parameter K described by the Gompertz equation. We have found that coefficient D is directly proportional to the growth parameter K . The fit curve significantly matches the empirical data ( P  < 0.05), with a correlation coefficient of 0.9152. Therefore, a transformed Gompertzian growth function was obtained accordingly to D . The degree of correlation between the Gompertzian growth parameter and the coefficient D allows a new interpretation of the growth parameter K on the basis of morphological measurements of a set of tumour cell types, supporting the idea that cell-growth kinetics can be modulated by phenotypic organization of attached cells.     

The relationship between kinetics of substrate-limited transitions and steady-state growth in continuous cultures ofAquaspirillum autotrophicum limited by pyruvate

Heterotrophic growth at steady state and during transient states caused by the sudden change of the concentration of the limiting factor in the feed medium was investigated experimentally for continuous cultures ofAquaspirillum autotrophicum limited by pyruvate. A model for describing the growth at steady state was selected from three unstructured models after statistical tests of the data. This model postulates that the growth yield increases linearly with the growth rate. Growth during transitions where the substrate remained limiting at all times was fitted with first-order kinetics. Theoretical predictions of these kinetics were derived from the unstructured models used to describe steady state. The predicted rate coefficients of the transients were compared to the experimental coefficients. It appeared that the model which best described steady-state growth also provided the best predictions for growth during the transient state. It is a widespread opinion that unstructured models are adequate to describe growth under steady-state conditions but not to predict transitions in continuous culture. However, for the particular case studied here, no higher degree of complexity was required to describe transitions, provided the growth of the culture was always limited by the substrate.     

Characteristic Species Dependent Growth Patterns of Mammalian Neoplasms

Evidence is presented, arising from an analysis of published data on tumour growth in three species of laboratory animals and in human multiple myeloma supporting a species specific relation between two supposedly independent parameters in the Gompertz equations frequently used to quantify tumour growth curves. This evidence supports the conjecture of Norton et al. (1976), based on their observations of the growth kinetics of a murine melanoma and a rat mammary carcinoma, that such a relation may be a general feature of tumour growth. Published data on the growth of xenografts of human colorectal tumours in immune-deprived mice suggests that the observed growth relation reflects the ability of a particular species to support a tumour of a certain maximum size. the existence of this relation greatly simplifies the task of predicting complete patterns of undisturbed neoplastic growth in these species.     

Alterations In Gastrointestinal Steady-State Kinetics Associated With the Growth of Experimental Tumours

Changes in the growth kinetics of the intestinal epithelium were observed in mice bearing the Lewis lung carcinoma and the T1699 mammary adenocarcinoma and in rats bearing the H-4-II-E₂ hepatoma. Proliferative activity in the jejunal tissue was markedly depressed with increasing tumour burden. Simultaneously, a significant reduction in total crypt cellularity occurred, followed by a reduction in villus height. While the total number of proliferative cells per crypt decreased, the relative proliferative compartment within the shrinking crypt increased. the rate of mucosal DNA synthesis remained constant during the initial cytokinetic changes, falling only after proliferative activity of the intestine was reduced to less than 50% of control levels. No general correlation could be drawn from the three tumour models studied between the level of gastrointestinal proliferation and tumour size, tumour growth rate or loss of weight by the tumour-bearing animals. However, intestinal proliferation was reduced by 50% when the tumour burden for each of the three tumours reached 6–8% of the host animal weight.     

Early spontaneous immortalization and loss of plasticity of rabbit bone marrow mesenchymal stem cells

Objectives: Bone marrow‐derived mesenchymal stem cells (BM‐MSC) have been widely used for cell therapy and tissue engineering purposes. However, there are still controversies concerning safety of application of these cells after in vitro expansion. Therefore, we aimed to investigate the characteristics of rabbit BM‐MSC during long‐term culture. Materials and methods: In this study, we have examined growth kinetics, morphological changes, differentiation potential and chromosomal abnormalities, as well as tumour formation potential of rabbit BM‐MSC in long‐term culture. Results and conclusion: We found that shortly after isolation, proliferation rate of rabbit BM‐MSC decreases until they enter a dormant phase. During this period of quiescence, the cells are large and multinucleate. After some weeks of dormancy we found that several small mononuclear cells originated from each large multinucleate cell. These newly formed cells proliferated rapidly but had inferior differentiation potential. Although they were immortal, they did not have the capability for tumour formation in soft agar assay or in nude mice. This is the first report of spontaneous, non‐tumorigenic immortalization of BM‐MSC in rabbits. The phenomenon raises more concern for meticulous monitoring and quality control for using rabbit BM‐MSC in cell‐based therapies and tissue engineering experiments.     

Temperature dependence of development rate and adult size in Drosophila species: biophysical parameters

Adult size in Drosophila results from the ratio of the rate of biomass increase and the rate of differentiation, both rates being temperature sensitive. Data on rates and size are collected in two tropical and two temperate Drosophila species; differentiation rate is higher in the two tropical species, growth rate differs between the large and small species of similar climatic origin. A biophysical model is used to evaluate the temperature dependence of adult size in Drosophila. The model is based upon the Sharpe–Schoolfield equation connecting enzyme kinetics and biological rates. Temperature sensitivities of growth rate, development time, and wing and thorax size are characterized by biophysical parameters. The biophysical parameter indicating trait specific temperature sensitivity is lower in tropical species than in temperate species, both for growth rate and for differentiation rate. In the larger species of a climate pair, thorax size and wing size prove to differ in pattern of temperature dependence; in the smaller species of a geographical pair, thorax size and wing size have identical patterns of temperature dependence.     

SYNERGISTIC REGULATORY EFFECTS OF TNFα, IL-1αAND IFNα ON THE GROWTH AND DIFFERENTIATION OF DAUDI LYMPHOMA CELLS

The regulatory effects of the combined treatment of tumour necrosis factorα (TNFα), interleukin-1α (IL-1α) and interferonα(IFNα) on the growth and differentiation of Daudi lymphoma cells were investigated. By means of anti-BrdU monoclonal antibodies and [³H-thymidine] incorporation a reduced proliferation rate was shown both through a combi-nation of TNFα with either IL-1α or IFNα and, above all, through simultaneous treatment with the three cytokines. In parallel, the degree of differentiation was evaluated via morphological criteria and detection of Fc receptors (FcR) and appeared higher after treatment with the three cytokines. Our results provide evidence of the increased sensitivity of this cell line to this combined treatment supporting the existence of a synergistic interaction in inducing the antiproliferative and differentiative effects.     

Somatic embryogenesis and indole alkaloid production in Catharanthus roseus

ABSTRACT

A new protocol to obtain an embryogenic cell line from cultured seedling explants of Catharanthus roseus is described. In order to assess the relationship between tissue differentiation and secondary metabolite biosynthesis, the biosynthetic capabilities (alkaloid production) of an embryogenic cell line and two non-embryogenic C. roseus strains were comparatively examined. Faster cell growth rate was associated with higher alkaloid production in the embryogenic cell line. The kinetics of ajmalicine and serpentine production by the three cell lines is also reported.     

Modeling of growth kinetics for <Emphasis Type="Italic">Pseudomonas putida</Emphasis> during toluene degradation

Glucose has been often used as a secondary substrate to enhance the degradation of primary substrate as well as the increase of biomass, especially for the inhibitory range of substrate concentration. In this study, we investigated the effect of glucose concentration on growth kinetics of Pseudomonas putida during toluene degradation for a wide concentration range (60–250 mg/l). Batch microcosm studies were conducted in order to monitor bacterial growth for three different initial concentrations (2, 5, 10 mg/ml) of glucose for a given toluene concentration. Modeling of growth kinetics was also performed for each growth curve of glucose dose using both Monod and Haldane kinetics. Batch studies revealed that bacterial growth showed a distinct inhibitory phase above some limit (∼170 mg/l) for the lowest (2 mg/ml) glucose dose, but the degree of inhibition decreased as the glucose dose increased, leading to three different growth patterns. The bacterial growth followed each of the modified Wayman and Tseng, Wayman and Tseng, and Luong model as the glucose dose increased from 2 to 10 mg/ml. This indicates that glucose has a prominent influence on bacterial growth during toluene degradation and that different kinetics should be adopted for each broth condition.     

CELL PROLIFERATION IN WALKER TUMOUR GROWING IN THE STOMACH WALL

Tumour cells from a Walker carcinosarcoma 256 were implanted in the gastric mucosa in rats. The tumour grew and infiltrated the lamina propria and the submucosal space after 7 days. It appeared to grow faster in the submucosal space than in the lamina propria. The cell proliferation was therefore studied separately in: (1) the tumour in the lamina propria, (2) the main tumour mass and (3) the tumour periphery, defined as the cells located within the outer 100–120 μm of the tumour. Mitoses arrested with vinblastine, cells labelled with tritiated thymidine and the grain count per labelled cell were studied at the three different sites. The rate of cell proliferation in the tumour was highest in the lamina propria, lower in the centre of the main tumour mass, and lowest at the periphery. Cell loss might explain the discrepancy between the rate of cell proliferation and the actual tumour growth. The factors that influence tumour cell proliferation in the different parts of the tumour are discussed.     

Effect of ammonium supplementation on production of poly-B-hydroxybutyric acid byAlcaligenes eutrophus in batch culture

Summary Alcaligenes eutrophus H16 showed some degree of oxygen sensitivity, having the fastest specific growth rate at a dissolved oxygen tension of 60% air saturation. Growth and PHB accumulation kinetics were studied and an accelerated rate of PHB synthesis was obtained when a low concentration of NH₄Cl was supplied during the PHB synthesis stage.     

Bioreactor studies on the effect of dissolved oxygen concentrations on growth and differentiation of carrot (Daucus carota L.) cell cultures

A bioreactor control system was used to investigate the effects of two dissolved oxygen concentrations (10% and 100%) on the growth and differentiation of Daucus carota L. cell cultures. The strategy used allowed the dissolved oxygen concentration to be controlled without the need for changing either the agitator speed or the total gas flow rate. During the proliferation phase, reducing oxygen resulted in a lower growth rate and in a delay in sugar uptake kinetics. Nonetheless, varying levels of oxygen were observed to have no effect on the final dry biomass. The higher alcohol dehydrogenase activity obtained under reduced oxygen conditions suggests that proliferating cultures adapted to the hypoxic environment by inducing alcoholic fermentation. Cell differentiation was highly sensitive to reduced oxygen since under this condition, the somatic embryo production was inhibited by about 75%. Sugar uptake and embryo formation were also delayed.Abbreviations ADH alcohol dehydrogenase - 2,4-D 2,4-dichlorophenoxyacetic acid - DO2 dissolved oxygen - SE somatic embryos - Tris tris(hydroxymethyl)-aminoethane     

Morphogenesis and growth kinetics of <Emphasis Type="Italic">Fusarium verticillioides</Emphasis>

The present study aimed to analyze the growth kinetics and morphogenesis of toxigenic Fusarium verticillioides strains. Growth curves based on mycelial dry weight measured after 24, 48, and 96 h and every 4 days for a period of 60 days were obtained for each strain. The morphogenesis of this fungus was studied during its developmental stages through the fluorescent method Fluorecein Diacetate-FDA and Ethidium Bromide-EB. The growth curves of the strains usually followed a homogeneous pattern comparable to the ideal growth curve. Using the fluorescence method, non-viable cells showed a light bright red coloration and viable cells presented green fluorescence with three fluorescent patterns. The present results showed that the morphogenesis of F. verticillioides is an asynchronous process characterized by the presence of a wide variety of fungal forms until 50 days of culture. The method is very useful to demonstrate the F. verticillioides growth stages as well as the perfect differentiation between viable and non-viable cell.     

Inhibition of growth and spontaneous metastasis of syngeneic transplantable tumors by an aromatic retinoic acid analogue

Summary Aromatic retinoic acid analogues selected for their favourable therapeutic ratios were tested for their effects on the growth (in vivo and in vitro) and spontaneous metastasis of a variety of murine sarcomas and carcinomas. Ro 10-1670 (a trimethylmethoxyphenyl analogue of retinoic acid) was used in in vitro studies, while its corresponding ethyl ester Ro 10-9359 was used for oral administration. Four of six fibrosarcomas, one of three squamous cell carcinomas, and one of five mammary adenocarcinomas responded to retinoid treatment in vivo by reduced growth rates (first detectable after 8–10 days), and in some cases by complete regression. The magnitude of the response was directly proportional to tumour immunogenicity, and eight tumours which failed to respond to retinoids did not evoke detectable transplantation immunity in syngeneic recipients. Retinoid administration did not significantly inhibit the development of spontaneous metastasis of non-immunogenic tumours, but decreased the incidence of secondary disease in the case of tumours of moderate immunogenicity. In vitro, retinoid treatments were generally without significant effects on tumour cell growth rate or morphology, and where growth inhibition was obtained it did not correlate with in vivo tumour responsiveness. No evidence of increased differentiation of retinoid-treated tumours was obtained either in vitro or in vivo. Taken together, the data suggest that in the 14 transplantable syngeneic tumours studied the inhibitory effects of retinoids on tumour growth and metastasis in vivo were mediated indirectly by potentiation of cell-mediated immunity directed against antigenic determinants on the tumour cell surface.     

Antidiabetic AD4743 enhances adipocyte differentiation of 3T3 T mesenchymal stem cells

AD4743 is an antidiabetic agent that, when added to fetal bovine serum (FBS), has been shown to have adipogenic activity for some proadipocyte cell lines once they reach confluence. In the current study, the effects of AD4743 on the growth and adipocytic differentiation of 3T3 T multipotential mesenchymal stem cells have been tested. 3T3 T cells, unlike other cells capable of undergoing adipocyte differentiation, are routinely induced to differentiate at low cell density. This is done using platelet-poor human plasma (HP), a potent inducer of growth arrest and differentiation. AD4743 (0-200 micrograms/ml) was tested in varied concentrations of HP or FBS, at varied cell densities, and at various times during growth and differentiation. AD4743 slowed the growth rate of 3T3 T cells and it induced their differentiation in a dose-dependent manner in medium containing 10% FBS once they reached confluence. The data suggest that the ability of AD4743 to inhibit growth may also be coupled with its ability to enhance differentiation. In addition, AD4743 (1-10 micrograms/ml) in the presence of 25% HP markedly increased the kinetics of adipocyte differentiation, at low (less than 5,000 cells/cm2) or high cell density. Greater than 50% cell differentiation could be achieved in 2 days in low density cultures; 80-95% differentiation could be achieved in just 4 days, compared to 8-12 days in a typical culture. The maximum amount of differentiation in HP was potentiated by AD4743 to a greater degree in poor lots of HP; however, the kinetics were increased in all lots. Adipocytic differentiation was measured both morphologically and by Northern blot analyses of differentiation-specific genes. AD4743 at 1-10 micrograms/ml appeared to be most effective, depending on the cell density and other conditions. The mechanism of action of AD4743 remains to be elucidated, but the enhancement of adipocyte differentiation does not appear to occur via an insulin-dependent pathway.     

Chlorophyllous Nature and Growth Characteristics of Teratoma and Habituated Tissue Cultures of Tobacco

The growth characteristics of three types of tissue cultures of tobacco have been investigated. They are (i) pigmented and differentiated teratoma tissue; (ii) pigmented and non-differentiated habituated tissue and (iii) non-pigmented and non-differentiated habituated tissue. All the tissues were grown on a 10 × White's medium without any exogenous supply of auxin. The comparative growth rate of the tissues was related to their nature of differentiation, viz., either organelle differentiation with chloroplast or organ differentiation with buds, leafy appendages, etc. Teratoma and pigmented habituated tissues had normal chlorophyll and carotenoid contents and showed more favorable growth in light than in the dark. The pigmented tissues showed a higher protein content than the non-pigmented tissues irrespective of whether they were of tumorous or normal origin. The pigmented tissues showed less dependence on sucrose availability in the medium than the non-pigmented tissues. Recovery after sucrose starvation was higher in the pigmented tissues than in the non-pigmented tissues. The results indicate that the growth rate of highly differentiated tissues is much slower than non-differentiated tissues and that the pigmented tissues photosynthesize and survive on reserve food products manufactured as a result of photosynthetic activity.     

IN VITRO PROLIFERATION OF MOUSE LYMPHOBLASTOID CELL LINES: GROWTH MODULATION BY VARIOUS POPULATIONS OF ADHERENT CELLS

The in vitro growth pattern of a number of mouse lymphoblastoid tumour cell lines was modified in the presence of adherent cell layers from various sources. The AVRij-1 and ST-4b cell lines exhibited a concentration—dependent growth pattern, i.e., they would only grow well when seeded at high starting cell concentrations. Better growth of these cells from low cell concentrations was observed in the presence of adherent cell layers from syngeneic or allogeneic bone marrow. Adherent cell layers derived from mouse spleen and pleural or peritoneal cavity could also promote the growth of the above tumour cells, but in a narrower range of cell concentrations and to a lower extent. Moreover, confluent adherent layers from the pleural and peritoneal cavities completely inhibited the growth of AVRij-1 and ST-4b cells, while adherent cell layers from the bone marrow did not inhibit growth at any cell concentration tested. The in vitro growth of concentration—independent cell lines was also affected by the presence of adherent cells from the bone marrow. Under syngeneic conditions, a slight increase in the growth of the ‘null’ or pre-B lymphoma cell line ABLS-8.1 was observed. On the other hand, the growth of tumour cells expressing more differentiated properties, such as the thymus T lymphoma tumour cell line ST-1.3 and the plasma cell tumour MPC-11.45.6.2.4, was inhibited in the presence of syngeneic bone marrow derived adherent cell layers. This inhibition was more pronounced under allogeneic conditions. Growth inhibition was also observed when concentration—independent cell lines were co-cultured with adherent cells from the pleural and peritoneal cavities. Thus, adherent cell layers from non-haemopoietic sources inhibited the growth of all cell lines tested. On the other hand, adherent cells from the bone marrow had a differential effect on growth of lymphoblastoid tumour cell lines. This depended on the in vitro growth properties of each tumour cell line and on some additional specific tumour cell properties. The latter could relate to the differentiation stage characterizing each tumour cell line. The culture method described here may serve as a model system for studies on interaction of leukaemic cell and the haemopoietic microenvironment.