Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Spheroids, Cellular”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 37 records · Page 2Linked to original sources
PubMed1995

Tumour fragment spheroids from human non-small-cell lung cancer maintained in organ culture.

Biopsy material from 17 human non-small-cell lung carcinomas (NSCLC) was maintained in agar overlay culture as tumour fragment spheroids for 40 days. A practical procedure for the formation of spheroids and organ culture is described. The mechanically dissociated tumour specimens showed a variation in their ability to generate spheroids that was not related to the ploidy or the histological differentiation of the biopsies. Light microscopic observations revealed a heterogeneous spheroid population with a mixture of tumour cells and stromal elements. Most of the histological elements normally found in human NSCLC could be seen in the spheroids. The cellular components in the spheroids varied between highly cellular to sparsely cellular, dominated by stromal elements. The squamous carcinomas were in general found to generate highly cellular spheroids more often than the adenocarcinomas. Spheroids with a different cellular content could be selected in vitro by using a morphometric technique. Diameter measurements showed a large variability in spheroid growth. Most of the spheroids decreased in size although bromodeoxyuridine labelling indicated active cell proliferation in the specimens. Frequent changes of medium did not affect spheroid growth. The culture system presented provides a model for studying the cellular heterogeneity as well as the biological characteristics of tumour tissue from individual patients in vitro.

Aged↗
PubMed1987

Influence of glucose and buffer capacity in the culture medium on growth and pH in spheroids of human thyroid carcinoma and human glioma origin.

pH gradients were measured with microelectrodes in cellular spheroids of human glioma (U-118 MG) and thyroid carcinoma (HTh7) origin. pH decreased outside the spheroids and then continuously decreased when the electrode was moved through the spheroid towards the center. The lowest pH values inside the spheroids were in the range 6.7-6.8 when grown under standard conditions with F10 medium. When medium with stronger buffer capacity (Dulbecco's minimum essential medium or Locke's) was used, the gradients in both types of spheroids were less steep. Less steep pH gradients were also obtained in both types of spheroids when the concentration of glucose was lowered to 0.1 g/liter in the medium. In the case of HTh7 spheroids the low pH inside the spheroids under standard culture conditions seemed toxic because the growth rate increased when the spheroids were cultured under conditions giving higher central pH values (high buffer capacity or low glucose concentration). No such growth-stimulating effects could be seen for the U-118 MG spheroids. The growth rate of both types of spheroids was retarded when they were grown in medium with very high glucose concentration (10 g/liter). The thickness of the viable cell layer increased for HTh7 spheroids when the concentration of glucose was lowered to 0.1 g/liter. A decrease in the thickness of the viable layer of U-118 MG spheroids was observed when they were grown at a high glucose concentration (10 g/liter).

Acid-Base Equilibrium↗
PubMed1992

Relations between the penetration, binding and average concentration of cytostatic drugs in human tumour spheroids.

A penetration assay based on freeze-drying and vapour fixation was applied to show the spatial distribution of non-bound and bound cytostatic drugs in cellular spheroids. Several studies have proposed that peripheral binding of drugs correlates with limited penetration. We showed that granular accumulation, mainly at the peripheral part of spheroids, might occur in parallel with good penetration. For example, this was the case in human glioma spheroids after incubation with Adriamycin for 15-30 min. Following treatment with actinomycin D, colon carcinoma spheroids exhibited rather good penetration but also showed granular accumulation mainly in their peripheral regions. Ara-C accumulated largely and homogeneously in the peripheral regions of colon carcinoma spheroids and this severely delayed penetration. It took about 1 h for ara-C in the central regions of the spheroids to reach the same concentration as in the culture medium. In contrast, ara-C easily penetrated glioma spheroids without accumulating noticeably at the periphery. Retention tests involving washing and further incubation in drug-free culture medium revealed that the areas demonstrating extensive accumulation most often retained the drug, indicating binding, whereas the concentration of drug in other areas decreased. The oil-centrifugation method, which was used for rapid separation of the spheroids from the drug-containing medium, showed that the average concentration of daunomycin in the spheroids exceeded that in the culture medium as early as after 15 min, by which time only limited penetration had occurred. We found that good penetration of ara-C correlated with a low average concentration in glioma spheroids, whereas limited penetration correlated with a high average concentration in colon carcinoma spheroids. The latter finding was attributable to the high accumulation of drug at the spheroid periphery. Thus, there was an inverse relationship between penetration and binding and between penetration and average drug concentration. It seemed that binding delayed or prevented penetration, whereas little, if any binding resulted in better penetration. Granular binding such as that observed Adriamycin and actinomycin D gave intermediately good penetration.

Antineoplastic Agents↗
PubMed1987

Response of spheroids implanted in the peritoneal cavity of mice exposed to cyclophosphamide and ionizing radiation.

Chinese hamster V79-171B spheroids implanted in the peritoneal cavity of C3H mice were characterized as a model for evaluating the toxicity of drugs requiring metabolic activation in vivo. After 24 hours in the peritoneal cavity, spheroid cellularity and plating efficiency were not significantly decreased, and host cell infiltration was estimated between 5 and 10%. The oxygenation of spheroids in the peritoneal cavity was assessed using their response to ionizing radiation. Spheroids were recovered after irradiation, incubated for 20 minutes in vitro with the slowly penetrating fluorescent dye, Hoechst 33342, and reduced to single cells with trypsin. Cells were analysed for clonogenicity as a function of position within the spheroid by using fluorescence-activated cell sorting in conjunction with the Hoechst diffusion gradient. When spheroids were first placed in the peritoneal cavity, the hypoxic fraction was close to 100%, but after 24 hours, cell oxygenation increased, probably due to a decrease in cell respiration rate. However, the location of a spheroid within the peritoneal cavity did not influence the radiosensitivity of individual spheroids or the amount of Hoechst 33342 delivered to the spheroid when Hoechst was given intravenously; individual spheroids recovered from mice given an intravenous injection of Hoechst showed no greater heterogeneity in binding than that observed when spheroids were incubated with Hoechst in vitro. Mice implanted with spheroids were also exposed to cyclophosphamide; the external cells of 0.6 mm diameter spheroids were about 30% more sensitive than the internal cells to the toxic effects of both cyclophosphamide and X-rays, and the combination of the two agents was additive at all depths within the spheroid.

Animals↗
PubMed1990

Damage to multicellular human H-2 glioma spheroids incubated with LAK cells: an ultrastructural study.

Local brain tumor therapy using lymphokine-activated killer (LAK) cells and recombinant interleukin-2 (rIL-2) has not proved to be effective in preliminary clinical trials. One obstacle to effective use of this therapy is ignorance about the events that follow contact of the LAK cells with glioma tissue. We used multicellular spheroids grown from human glioma cell lines as targets to study, in vitro, the effect of LAK cells against three-dimensional glioma tissue. Here we describe the ultrastructural changes in spheroids of H-2 glioma cells incubated in pellets of LAK cells for up to 24 hours. In H-2 spheroids, cellular damage was not restricted to the effector cell-target cell (effector-target) contact; it extended farther, at least partly because of nonspecific changes in the spheroid micromilieu. Formation of cytoplasmic blebs, a characteristic effect of T cells, natural killer cells, and LAK cells on single target cells, also occurs in H-2 spheroids, and it is not limited to the effector-target contact area either. These findings suggest that LAK cells release membrane-damaging agents that remain active outside the effector-target area, in the micromilieu of H-2 spheroid tissue.

Cytotoxicity, Immunologic↗
PubMed1992

Activity of the human metallothionein promoter (hMT-IIA) in cell populations isolated from varying depths in multicell spheroids following exposure to cadmium.

In order to evaluate whether tumor microenvironment might influence the response of the metallothionein promoter to heavy-metal exposure, we transfected HT-29 colon carcinoma cells with the vector phMTIIA-CAT-neo, containing a fusion gene consisting of 426 bp of the human metallothionein-IIa (hMT-IIA) promoter immediately upstream of the bacterial chloramphenicol acetyl transferase (CAT) gene. We grew one of the stable transfectants (clone 20) as three-dimensional multicell tumor spheroids, exposed them to CdCl2 and measured CAT expression in cells isolated from various depths into the spheroids. Cellular populations were isolated by flow cytometry on the basis of Hoescht 33342 fluorescence intensity, taking advantage of the dye's diffusion gradient to isolate cells from inner (dim) and outer (bright) regions of stained spheroids. When intact spheroids were incubated for 18 h in the presence of 5 microM CdCl2, CAT activity was induced in all cell fractions isolated from the spheroids, but induction was 10-fold greater in cells in the outermost fraction (fraction 10) than inner fraction (fraction 2). When spheroids were dissociated, sorted into individual fractions and then incubated with cadmium, CAT expression was maximized in all fractions. Exposure of intact spheroids to 30 microM CdCl2 resulted in increased CAT induction in cells isolated from the internal fractions of the spheroids. The data suggest that limited diffusion of cadmium through cells organized in a tissue-like arrangement may account for the lower levels of hMT-IIA promoter activity observed in cells collected from increasing depths into the spheroids.

Cadmium↗
PubMed1995

Further application and evaluation of critical cell number and modifying factors in radiocurability of multicellular spheroids.

Relationship of cure and surviving clonogenic cell number after various doses of X-irradiation was examined in multicellular spheroids of LCT1 human lung adenocarcinoma cells, LCT2 human lung small cell carcinoma cells and FSA1233 mouse fibrosarcoma cells. Some of these spheroids were cured at such doses that considerable number of clonogenic cells still survived after irradiation. Radiocurability was analyzed by comparing total clonogenic cell number in spheroids, cellular radiosensitivity and critical cell number Nc, i.e., the minimum number of clonogenic cells required to produce regrowth. (Nc-1) cells were killed by post-radiation processes and the larger the critical cell number, the more radiocurable. The LCT2 spheroids had the largest critical cell number and were most radiocurable. To investigate underlying mechanisms, modifying effect of heavily irradiated (HIR) tumor cells on the clonogenicity, i.e., plating efficiency of unirradiated tumor cells was investigated. Plating efficiencies with HIR cells showed significant decrease in LCT2 cells, no change in LCT1 cells and increase in FSA1233 cells. The results indicated that in case of LCT2 spheroids some of viable cells surrounded by dying or dead cells might have been killed with unknown toxic effect additional to direct irradiation effect. Thus, critical cell number analysis may be useful to quantify and to compare modifying effect of cellular/environmental factors in curing process of spheroids or tumors.

Adenocarcinoma↗
PubMed1996

Options of flow cytometric three-colour DNA measurements to quantitate EGFR in subpopulations of human bladder cancer.

Flow cytometric multi-parameter analysis has proven to be a powerful tool to characterize subpopulations of cell suspensions, and is applied routinely in hematology. However, in studies of cancer where there is interest in defining phenotypic markers in conjunction with DNA content, this method has hardly been applied [6]. Our objective was to develop a methodology that extends previous investigations on relative and absolute quantitation of the epidermal growth factor receptor (EGFR) in dual parameter analysis in vitro on human bladder cancer cell lines [3]. In order to quantitate EGFR content in tumours and to relate it to DNA content, tumour selection, DNA-content, and EGFR-content measurements should be carried out simultaneously. Different fluorescent dyes were used to optimize DNA assessment and antibody staining, using a single laser instrument as a practical approach for clinical routine. In vitro cultures were used to validate the quality of tumour cell selection and antigen quantitation. Therefore, two urothelial tumour cell lines-lowly and highly differentiated-were incubated under different conditions: monolayer (ML), three-dimensional multi-cellular spheroids (MCS) and cocultures (COCU) with the fibroblast cell line NI were investigated and EGFR quantitation was related to S-phase fraction (SPF). Accurate determination of instrument settings allows simultaneous three-colour analysis with DNA assessment. Tumor cell selection based on staining with phycoerythrine (R-PE) against a highly expressed urothelial glycoprotein, detected with the antibody Uro5 or against cytokeratin appeared to be possible in FL2, using the fluorochrome combination fluorescein-isothiocyanate (FITC), R-PE and propidiumiodide (PI). Using this staining protocol, relative and absolute EGFR quantitation (quantum simply cellular beads) is shown to be accurate, when FITC is used for EGFR staining and measured in the green fluorescence channel (FL1). Using this colour combination EGFR content and SPF of tumour cells were compared in different growth states, and could be monitored reliably. In spite of a higher emission spectrum of 7-aminoactinomycin-D (7-AAD), this DNA stain provided no advantage over PI. Broad coefficients of variation (CV) were found when intact cells were stained, thus hindering accurate assessment of ploidy and S-phase fraction. Similarly, Syto-13, a DNA dye detected in FL1, could not be optimized for multi-parameter measurements. Although the emission maximum is at 520 nm, the spectrum is too wide to compensate fluorescence overlap in FL2 or FL3. Quantum Red (QR), used as a streptavidin conjugate in FL3, could not be combined with two other colours for DNA staining, since sufficient compensation was not obtainable when an argon ion laser is used. The coculture model allows verification of tumour cell selection and discrimination. The high differentiated tumour cell line RT4 shows an unambiguous correlation between EGFR content and S-phase fraction. The low differentiated tumour cell line J82 presents a similar pattern of post-transcriptional EGFR regulation with respect to culture condition, however, the S-phase fraction is basically unaffected.

Carcinoma↗
PubMed1997

Human prostate tumor angiogenesis in nude mice: metalloprotease and plasminogen activator activities during tumor growth and neovascularization of subcutaneously injected matrigel impregnated with human prostate tumor cells.

BACKGROUND: A critical aspect for growth of solid tumors is the development of a blood supply. Our objective was to establish a model for the study of angiogenesis of human prostate tumors by examining the growth of microvessels into Matrigel containing human prostate tumor cells implanted subcutaneously in nude mice. METHODS: Human prostate tumor cell lines PC-3 and LNCaP were injected in Matrigel under the abdominal skin of nude mice and were harvested at 4, 8, and 14 days post-injection. The growth of tumor cells and blood vessels was examined histologically and by immunohistochemical localization of von Willibrand Factor VIII (vWF). Since plasminogen activators and matrix metalloproteases are associated with angiogenesis, the activities and molecular forms of these proteases were determined in Matrigel control and Matrigel-tumor cell subcutaneous implants. RESULTS: Blood vessel formation in the Matrigel implants containing LNCaP and PC-3 cells was demonstrable at 8 days post-injection. However, the pattern of blood vessel formation by the two tumor cell lines was different; PC-3 tumors showed a more invasive phenotype and smaller diameter blood vessels, whereas LNCaP tumors grew as large cellular spheroids surrounded by large, dilated blood vessels. Many blood vessels of PC-3 tumors expressed vWF by day 14 of growth, whereas most blood vessels in LNCaP tumors were immunohistochemically negative for this antigen. Mouse skin blood vessels in the same PC-3 and LNCaP tumor histological sections were positive for vWF. Matrigel contained both plasminogen activator and metalloprotease activities. The plasminogen activator activity in Matrigel control implants was totally inhibited by 4 days post-injection, indicating the presence of an inhibitor provided by the host mouse. LNCaP tumor cells injected did not have appreciable plasminogen activator activity, nor did LNCaP tumors develop plasminogen activator activity with tumor growth post-injection. PC-3 cells did have plasminogen activator activities, which were partially negated after subcutaneous injection (4 days), but then increased again by 8 days post-injection. This increase in plasminogen activator activity was due to urokinase (about 54 kDa) produced by the tumor and not by the mouse host (mouse urine urokinase about 44 kDa). Matrigel alone demonstrated gelatinase B (about 95 kDa) activity in zymograms, and gained considerable gelatinase A (about 70 and 74 kDa) activity after subcutaneous implantation. No metalloprotease activity from the tumor cells could be distinguished over that contributed by the mouse host cells in the Matrigel. Matrigel also contains caseinolytic activities of approximately 56, 80, 85, and 89 kDa. After subcutaneous injection of Matrigel, the 89 kDa form increases considerably in activity and the others are diminished. This pattern is also observed in LNCaP and PC-3 tumors post-injection, except the PC-3 tumors demonstrate increased 56 kDa activity. CONCLUSION: The subcutaneous growth of LNCaP and PC-3 prostate tumor cells in Matrigel in nude mice can be used to study tumor-induced angiogenesis. However, the organization of LNCaP and PC-3 tumor growth and the pattern of microvessels associated with each tumor are different in this system, implying that each tumor has unique influences on the pattern of microvessel development. The mode of action by which this is brought about is not known, but may be due to specific factors produced/ released by the tumor cells.

Animals↗
PubMed1984

Influence of migratory blood cells on the attachment of tumor cells to vascular endothelium.

The purpose of this study was to determine whether the presence of migratory blood cells in association with tumor emboli had the capacity to alter the attachment of tumor cells to vascular endothelium. Highly metastatic RT7-4bs rat hepatocarcinoma cells were labelled with [125I]UdR before being allowed to form mixed cellular spheroids incorporating resident peritoneal macrophages, activated peritoneal macrophages, polymorphonuclear leukocytes, splenic T lymphocytes, or splenic B lymphocytes derived from both normal and tumor-bearing animals. The presence of polymorphonuclear leukocytes or activated macrophages led to a considerable increase in the number of tumor cells attaching to endothelial cell monolayers in vitro. The presence of T or B lymphocytes from either normal or tumor-bearing rats was without effect on tumor attachment to endothelium. Increased tumor cell retention in the lungs was evident for mixed spheroids containing tumor cells and polymorphonuclear leukocytes compared to homotypic tumor spheroids composed of tumor cells alone. Furthermore, preinjection of polymorphonuclear leukocytes intravascularly or inoculation of tumor cells as heterotypic spheroids containing polymorphonuclear leukocytes increased lung colony formation over that obtained after inoculation of tumor cells alone. Several simple sugars were tested for their ability to block tumor cell, polymorphonuclear leukocyte or activated macrophage binding to endothelium in vitro. The results indicate that the glycosylated cell surface components mediating tumor cell attachment to endothelium are not identical with those mediating attachment of either polymorphonuclear leukocytes or activated macrophages. Medium conditioned during mixed spheroid formation was without effect on tumor cell attachment to endothelium. We conclude that the presence of some, but not all classes of leukocytes can modulate tumor cell attachment to vascular endothelium, an effect most likely mediated by a mechanism involving direct contact between the leukocytes and the endothelial cell monolayer.

Animals↗
PubMed1993

Bioreducible mustards: a paradigm for hypoxia-selective prodrugs of diffusible cytotoxins (HPDCs).

Existing hypoxia-selective cytotoxins (HSCs) are designed to kill only the hypoxic subpopulation in tumours, and to be used in conjunction with other therapies (e.g., radiation). A new class of drugs, hypoxia-activated prodrugs of diffusible cytotoxins (HPDCs) are proposed. These are designed to exploit, rather than merely deal with, tumour hypoxia, by releasing diffusible cytotoxins on bioreduction in hypoxic regions. Such diffusible cytotoxins are required to be much more cytotoxic than the parent prodrug, to be sufficiently stable (half lives from 0.1 to 10 min) to allow them to diffuse up to 200 microns from the hypoxic regions, and to be equally effective against all major tumour cell subpopulations, including non-cycling cells. Nitrogen mustards, which show little cell cycle specificity, which kill cells by a well-understood mechanism (DNA cross-links), and which have stabilities and reactivities able to be predictably controlled by structural variations, are proposed as suitable candidates fur such diffusible cytotoxins. Design parameters for two classes of potential HPDCs are discussed; nitro-deactivated aromatic mustards, and cobalt (III) complex-deactivated aliphatic mustards. Examples of both classes show greater cell-killing activity against intact compared with dissociated multi-cellular spheroids. This suggests they may indeed function as HPDCs, by penetrating to the hypoxic core of the spheroid and there releasing potent cytotoxins which diffuse out to kill surrounding cells at lower oxygen tensions.

Animals↗
PubMed1990

Expression of verapamil hypersensitivity in multidrug-resistant cells grown as multicellular spheroids.

It is known that certain multidrug-resistant cell lines are hypersensitive to verapamil and to some other membrane-active agents. We examined the expression of verapamil hypersensitivity in multidrug-resistant, verapamil-hypersensitive CHO cell line by cells grown in the form of multicellular spheroids. We found that although vincristine resistance is expressed at high levels in multicellular spheroids, we could not detect verapamil hypersensitivity when cells were grown in this form. Using a dry autoradiographic method to study the entry of [3H]-verapamil into the spheroids, we demonstrated that the lack of hypersensitivity was not due to incomplete drug penetration. We therefore propose that when hypersensitive multidrug-resistant cells are grown as three-dimensional spheroids, cellular interactions may modify the expression of verapamil hypersensitivity.

Animals↗
PubMed1988

In vitro and in vivo responses of a panel of murine colon tumours to TCNU: a positive correlation.

TCNU is highly active against a panel of three histologically distinct transplantable murine adenocarcinomas of the colon (MAC tumours). Significant reductions in colony formation (greater than 70%) were observed in vitro in all three cell lines following a 1 h exposure to TCNU at experimentally achievable drug C X t values. A good correlation exists between in vivo tumour responses and in vitro cell responses in all cases. Dose-response curves generated at increasing exposure times suggest that no active or long lived products of TCNU are formed as a result of the drug's spontaneous breakdown in vitro (rate of breakdown was 0.078 micrograms min-1 at 37 degrees C). Preliminary studies with the HT-29 human colon cell line have demonstrated that multi-cellular spheroids are more responsive to TCNU (2 X) than the same cells cultured as monolayers.

Animals↗
PubMed1993

Estramustine induces disorganization of microtubules, perinuclear retraction of vimentin and endoplasmatic reticulum, and inhibits cell migration.

The effects of the mitotic inhibitor estramustine on the cytoskeleton of DU 145 and AG 1518 cells were studied. Estramustine caused a partial disassembly of microtubules and withdrawal of microtubules from the cell periphery, disorganized microtubules and delayed regrowth of disassembled microtubules. It also induced a spheroid cellular morphology and affected cellular adhesion and survival. Sometimes microtubules seemed to be organized from several microtubule-organizing centers. The cytoskeleton-dependent cell migration was inhibited in the presence of estramustine and the microtubule-interacting vimentin and endoplasmatic reticulum retracted to the perinuclear area. Our results show that not only a complete disassembly of microtubules, but also disturbances of the microtubule network can have dramatic effects on microtubule-dependent processes and localization of cellular organelles. These effects could be of importance in the treatment of prostatic carcinoma with estramustine.

Cell Adhesion↗
PubMed2003

Galactosylated PVDF membrane promotes hepatocyte attachment and functional maintenance.

One of the major challenges in BLAD design is to develop functional substrates suitable for hepatocyte attachment and functional maintenance. In the present study, we designed a poly(vinylidene difluoride) (PVDF) surface coated with galactose-tethered Pluronic polymer. The galactose-derived Pluronic F68 (F68-Gal) was adsorbed on PVDF membrane through hydrophobic-hydrophobic interaction between PVDF and the polypropylene oxide segment in Pluronic. The galactose density on the modified PVDF surface increased with the concentration of the F68-Gal solution, reaching 15.4 nmol galactosyl groups per cm2 when a 1 mg/ml of F68-Gal solution was used. The adsorbed F68-Gal remained relatively stable in culture medium. Rat hepatocytes attachment efficiency on F68-Gal modified PVDF membrane was similar to that on collagen-coated surface. The attached hepatocytes on PVDF/F68-Gal membrane self-assembled into multi-cellular spheroids after 1 day of culture. These attached hepatocytes in spheroids exhibited higher cell functions such as albumin synthesis and P450 1A1 detoxification function compared to unmodified PVDF membrane and collagen-coated surface. These results suggest the potential of this galactose-immobilized PVDF membrane as a suitable substrate for hepatocyte culture.

Albumins↗
PubMed2004

Mathematical modelling of the spatio-temporal response of cytotoxic T-lymphocytes to a solid tumour.

In this paper a mathematical model describing the growth of a solid tumour in the presence of an immune system response is presented. In particular, attention is focused upon the attack of tumour cells by so-called tumour-infiltrating cytotoxic lymphocytes (TICLs), in a small, multicellular tumour, without necrosis and at some stage prior to (tumour-induced) angiogenesis. At this stage the immune cells and the tumour cells are considered to be in a state of dynamic equilibrium--cancer dormancy--a phenomenon which has been observed in primary tumours, micrometastases and residual disease after ablation of the primary tumour. Nonetheless, the precise biochemical and cellular mechanisms by which TICLs control cancer dormancy are still poorly understood from a biological and immunological point of view. Therefore we focus on the analysis of the spatio-temporal dynamics of tumour cells, immune cells and chemokines in an immunogenic tumour. The lymphocytes are assumed to migrate into the growing solid tumour and interact with the tumour cells in such a way that lymphocyte-tumour cell complexes are formed. These complexes result in either the death of the tumour cells (the normal situation) or the inactivation (sometimes even the death) of the lymphocytes. The migration of the TICLs is determined by a combination of random motility and chemotaxis in response to the presence of chemokines. The resulting system of four nonlinear partial differential equations (TICLs, tumour cells, complexes and chemokines) is analysed and numerical simulations are presented. We consider two different tumour geometries--multi-layered cell growth and multi-cellular spheroid growth. The numerical simulations demonstrate the existence of cell distributions that are quasi-stationary in time and heterogeneous in space. A linear stability analysis of the underlying (spatially homogeneous) ordinary differential equation (ODE) kinetics coupled with a numerical investigation of the ODE system reveals the existence of a stable limit cycle. This is verified further when a subsequent bifurcation analysis is undertaken using a numerical continuation package. These results then explain the complex heterogeneous spatio-temporal dynamics observed in the partial differential equation (PDE) system. Our approach may lead to a deeper understanding of the phenomenon of cancer dormancy and may be helpful in the future development of more effective anti-cancer vaccines.

Cell Division↗
PubMed2006

Monocyte chemoattractant protein-1 (MCP-1) inhibits the intestinal-like differentiation of monocytes.

Monocytes (MO) migrating into normal, non-inflamed intestinal mucosa undergo a specific differentiation resulting in a non-reactive, tolerogenic intestinal macrophage (IMAC). Recently we demonstrated the differentiation of MO into an intestinal-like macrophage (MAC) phenotype in vitro in a three-dimensional cell culture model (multi-cellular spheroid or MCS model). In the mucosa of patients with inflammatory bowel disease (IBD) in addition to normal IMAC, a reactive MAC population as well as increased levels of monocyte chemoattractant protein 1 (MCP-1) is found. The aim of this study was to investigate the influence of MCP-1 on the differentiation of MO into IMAC. MCS were generated from adenovirally transfected HT-29 cells overexpressing MCP-1, macrophage inflammatory protein 3 alpha (MIP-3alpha) or non-transfected controls and co-cultured with freshly elutriated blood MO. After 7 days of co-culture MCS were harvested, and expression of the surface antigens CD33 and CD14 as well as the intracellular MAC marker CD68 was determined by flow-cytometry or immunohistochemistry. MCP-1 and MIP-3alpha expression by HT-29 cells in the MCS was increased by transfection at the time of MCS formation. In contrast to MIP-3alpha, MCP-1 overexpression induced a massive migration of MO into the three-dimensional aggregates. Differentiation of IMAC was disturbed in MCP-1-transfected MCS compared to experiments with non-transfected control aggregates, or the MIP-3alpha-transfected MCS, as indicated by high CD14 expression of MO/IMAC cultured inside the MCP-1-transfected MCS, as shown by immunohistochemistry and FACS analysis. Neutralization of MCP-1 was followed by an almost complete absence of monocyte migration into the MCS. MCP-1 induced migration of MO into three-dimensional spheroids generated from HT-29 cells and inhibited intestinal-like differentiation of blood MO into IMAC. It may be speculated that MCP-1 could play a role in the disturbed IMAC differentiation in IBD mucosa.

Antigens, CD↗
PubMedDate not supplied

Analysis of tridimensional mixed cultures of mouse B16 melanoma cells and 3T3 fibroblasts.

The interactions between invasive malignant cells and normal fibroblastic cells were studied in a cellular spheroid model in vitro. Murine B16 melanoma cells (previously cultured in monolayer for a short period) and 3T3 mouse fibroblasts (greater than or equal to 130 passages in monolayers) were cultured under tridimensional conditions (pure or mixed spheroids). As compared to pure 3T3 or mixed spheroids, B16 spheroids were smaller and characterized by a higher proliferation rate, a lower degree of necrosis, and a less abundant extracellular matrix. Disintegration was observed in some pure 3T3 or mixed spheroids. Melanogenesis progressively increased inside B16 or mixed spheroids. By immunohistochemical methods and electron microscopy, laminin, fibronectin and collagen I, III and IV in extracellular matrix were studied in the three types of spheroids.

Animals↗