PubMed Health⌕ Search

SEARCH · PubMed Health

Results for “Diffusion Chambers, Culture”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original 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
PubMed1976

Growth of HeLa cells in diffusion chamber cultures in vivo.

In order to investigate some characteristics of Millipore diffusion chamber cultures (DCC) and their potential for the quantitation of tumor cell growth kinetics, HeLa S-3-cells were grown in diffusion chambers implanted i.p. in mice. HeLa cells grew pseudologarithmically in DCC with a population-doubling time of approximately 2.8 days when 9.2 to 12.0 X 10(5) cells were placed in the chambers initially. The population-doubling time varied as a function of the initial innocula size but was always longer than the in vitro population-doubling time of 1.2 days. Comparable harvests of cells from experiment to experiment varied quantitatively by no more than a factor 1.8. This variaton somewhat limits the use of DCC for the quantitation of subtle effects. The fate and morphology of HeLa cells in DCC were determined from scanning electron microscopy. DCC might be exploited in clinical studies of the growth of patients' tumors and the effects of various therapies in a host-mediated system.

Animals↗
PubMed1981

Appearance of B- or T-lymphocyte markers after diffusion chamber culture of acute lymphoblastic leukemia cells.

Blast cells derived from blood and marrow samples of a patient with acute lymphoblastic leukemia (ALL), as well as from the Reh line originally established from an ALL patient, were cultured in diffusion chambers implanted i.p. into preirradiated CBA mice. At different intervals over a period of up to 20 days, surface immunoglobulins, ALL antigen, and T-cell antigen were investigated by using direct immunofluorescence. Rosette formation was tested with sheep and mouse erythrocytes. On day 0, the cells expressed only ALL antigen at the surface, and no rosette formation was observed. During culture the patient's lymphoblasts, which originally had cytoplasmic IgM in addition to ALL antigen, expressed surface immunoglobulins as well as mouse erythrocyte receptors. The Reh line cells were ambivalent in two experiments developing T-cell antigens and sheep erythrocyte receptors as well as mouse erythrocyte receptors. Our data suggest that the differentiation arrest in leukemic lymphoblasts can be overcome, thus entailing a surface pattern similar to mature T- or B-lymphocytes.

Antigens↗
PubMed1976

Assessment of erythrocytic and granulocytic colony formation in an in vivo plasma clot diffusion chamber culture system.

Normal rat bone marrow cells seeded into a plasma clot diffusion chamber culture developed into erythrocytic and granulocytic colonies in vivo. Chambers implanted into the peritoneal cavity of normal hosts showed erythrocytic colony numbers reaching an initial peak on day 2, declining on days 3--5, and increasing in a secondary growth phase on day 7. Day 2 colonies were evenly dispersed; day 7 colonies were grouped into discrete areas of bursts. Granulocytic colony numbers reached a peak on day 4 and gradually declined through day 7. Cells in various stages of differentiation could be detected in both colony types. Colony numbers were proportional to the number of marrow cells seeded into the chamber. Host animals treated with phenylhydrazine induced a marked increase in erythrocytic colony numbers and size and a decrease in granulocytic colony formation. Host animals treated with endotoxin suppressed erythrocytic colonies while increasing granulocytic colony size. This method may prove advantageous for the study of hematopoietic colony formation in a physiologic environment.

Animals↗
PubMed1990

Human normal B lymphocytes produce a growth-promoting activity for granulocyte progenitors in diffusion chamber culture after stimulation with pokeweed mitogen.

We have measured the effect of normal B lymphocytes on more primitive granulocyte progenitors (CFU-dG) clonal growth in double-diffusion chamber culture in vivo. It was found that pokeweed mitogen (PWM) stimulated B cells produce a growth-promoting activity which augments the CFU-dG--derived myeloid colony formation in a dose-dependent fashion. All colonies formed under the experimental conditions were composed exclusively of granulocytes at different stages of maturation. Unstimulated B lymphocytes did not effect the CFU-dG clonal proliferation.

B-Lymphocytes↗
PubMed1985

Response of granulocyte-committed progenitors from patients with chronic myeloid leukemia to humoral regulators release by macrophages in agar diffusion chamber culture.

We investigated the responsiveness of granulocyte-committed progenitors (CFU-G/D) from patients with chronic myeloid leukemia (CML) and healthy subjects to stimulating and inhibiting activities released by murine macrophages in diffusion chamber culture. CFU-G/D from CML demonstrate a normal response to macrophage-derived stimulation. The responsiveness of CFU-G/D from patients with CML to indomethacin-sensitive inhibition was significantly suppressed. In this regard no difference between CFU-G/D from bone marrow and blood of patients with CML could be observed. Colonies formed both by CFU-G/D from healthy subjects and CML consisted exclusively of cells of granulocyte line: from myeloblasts up to polymorphonuclear granulocytes. Similar cellular composition of colonies could be noted during macrophage-derived stimulation and inhibition of CFU-G/D growth. In conclusion, we have demonstrated that CML CFU-G/D which proliferate and differentiate in diffusion chamber culture show a normal response to macrophage-derived stimulation but are less sensitive to indomethacin-dependent inhibition.

Adult↗
PubMed1985

Allocation of the suppressive activity of normal peripheral blood lymphocytes induced by diffusion chamber culture and Con A stimulation to the G2 phase of the cell cycle.

Peripheral blood mononuclear cells stimulated either by diffusion chamber culture or by a high Con A concentration exhibit suppressive activity under conditions where no increase in cell number takes place. Instead an accumulation of large cells is observed which, according to their DNA contents, are classified as cells in the G2 phase of the cell cycle. By elutriation separation the suppressive activity is shown to be confined to this cell cycle phase.

Adult↗
PubMed1982

Two classes of murine granulocyte progenitor cells expressed in plasma clot diffusion chamber cultures.

Two distinct classes of granulocyte progenitor cells present in normal mouse bone marrow are expressed sequentially in the vivo plasma clot diffusion chamber culture system. By several criteria, progenitor cells giving rise to granulocyte colonies on day 4 of culture (CFU-D4) are different from those giving rise to colonies on day 7 (CFU-D7). These differences include: cell cycle activity as measured by in vitro incubation with cytosine arabinoside, residual concentration in the bone marrow after in vivo treatment of donor mice with cytosine arabinoside or methotrexate, resistance to osmotic lysis, size as determined by velocity sedimentation, and the morphology of the granulocyte colonies to which these cells give rise. The CFU-D7 appears to represent an earlier progenitor cell than the CFU-D4 in the differentiation pathway of the granulocyte and is analagous in many respects to the BFU-E in the erythroid pathway.

Animals↗
PubMed1983

Chronic myelocytic leukaemia: cytogenetical studies on haemopoietic colonies and diffusion chamber cultures.

In 20 patients with chronic myelocytic leukaemia in the chronic phase or in blast crisis, several in vitro (CFU-C, BFU-E) and in vivo (diffusion chamber) culture techniques were used to demonstrate the existence of a still diploid cell clone. In 4 out of 12 patients in the chronic phase, Ph1-negative metaphases were found in the various cultures for haemopoietic progenitor cells but not in the standard suspension cultures used for cytogenetics. However, all the 8 patients investigated during blast crisis had only Ph1-positive and other abnormal clones, indicating the loss of the diploid clone during progression of the disease.

Acute Disease↗
PubMed1985

Mixed megakaryocytic-granulocytic differentiation during diffusion chamber culture of peripheral blast cells from the blast crisis of chronic myelocytic leukemia.

Morphologically and cytochemically undifferentiated peripheral blast cells from three patients in blast crisis of Ph'-positive chronic myelocytic leukemia were analysed morphologically, immunologically and cytogenetically prior to and during in-vivo diffusion chamber culture (DC) to investigate their differentiation capacity. In two patients immunological markers characteristic of the megakaryocytic lineage were found on the original cells, but lineage-specific differentiation markers were absent on the blast cells of the third patient. During DC culture multilineage differentiation capacity could be demonstrated immunologically and morphologically in all three patients with expression of megakaryocytic and granulocytic markers as well as terminal differentiation along these lineages. Cytogenetic analysis prior to and during DC culture provided evidence that the cells differentiating in culture were derived from the blast crisis clones and that the event leading to blast crisis might have occurred in a pluripotent precursor cell which retained its differentiation capacity along several lineages.

Adult↗
PubMed1995

Human bone tissue formation in diffusion chamber culture in vivo by bone-derived cells and marrow stromal fibroblastic cells.

Direct grafts of human cells into immunocompromised or cortisone-treated animals, either alone or within carrier materials, have been used with some success to assess the developmental capability of the grafted cells. However, identification of the donor or host origins of the generated tissue in such direct grafts is essential. In an alternative and extensively used experimental system, cells are cultured within the isolated environments of diffusion chambers, which are surgically implanted in appropriate hosts. This system allows the direct study of the cellular potentials for differentiation as host tissues are excluded. In the present study, human osteoprogenitor cell populations derived from trabecular bone explants or marrow suspensions of 3 patients (2 females aged 14 years and 1 male aged 27 years) were cultured in the absence or the continuous presence of dexamethasone (10 nmol/L). Cells were impregnated into porous hydroxyapatite ceramics before subcutaneous implantation, or placed within diffusion chambers before intraperitoneal implantation, in athymic mice. All subcutaneous implants of cells in ceramic showed morphological evidence for the formation of bone tissue. In the diffusion chambers it was found that both marrow- and bone-derived fibroblastic cells cultured in the absence of dexamethasone generally produced fibrous tissue only. When cultured in the continuous presence of dexamethasone (10 nmol/L), these cell populations produced similar osteogenic tissues with active osteoblasts, wide osteoid seams, and mineralized tissue, with cartilage toward the interior of the chamber. These results validate the diffusion chamber as an experimental system to study human osteogenesis using appropriately primed cell populations.

Adolescent↗
PubMed1985

Assessment of early hemopoiesis in in vivo diffusion chamber cultures.

Many studies have provided evidence that the cells which give rise to colonies containing granulocytes (CFU-DG) share several characteristics with other primitive hemopoietic precursors (CFU-S and CFU-MIX). Cell cycle status, sensitivity to hypotonic lysis, and velocity sedimentation rates are similar. On the other hand, CFU-DG can be distinguished from these precursors by difference in sensitivity to cryopreservation and hyperthermia. Uniqueness of the in vivo diffusion chamber culture system is accentuated by finding humoral activities produced by human HUT 102 and Mo T cell lines and murine BCL1 B cell line which seem to be different from the activities identifiable by CFU-C, BFU-E, and CFU-MIX assays in vitro.

Animals↗
PubMed1981

Expression of common acute lymphoblastic leukaemia antigen and terminal deoxynucleotidyl transferase in normal mononuclear blood cells during diffusion chamber culture.

Peripheral mononuclear blood cells of 2 healthy individuals were cultured over a 13 d period in diffusion chambers. A high percentage of cALL antigen positive cells and later on TdT containing cells appeared during culture. The cALL+ cells could be clearly distinguished into 2 groups: a population with weak and diffuse fluorescence and a small number of ring-like brightly dyed cells. The first group develops into TdT+ cells and therefore resembles immature T-cells. The interpretation of the ring-like stained cells is still preliminary, but it cannot be excluded, that these cells represent a cell population with more than lymphatic differentiation capacity.

Antigens, Neoplasm↗
PubMed2000

Granulocytopoiesis versus cytokine activities in peritoneal diffusion chamber cultures in the mouse.

Regulation of granulocyte formation was studied by correlating granulocytopoiesis in diffusion chambers with activities of putative regulatory cytokines in such chambers. The implantation procedure increased the levels in 1- and 2-d chambers of interleukin 6 (IL-6), G-CSF, TNF-alpha, and non-specific granulocyte/macrophage (G/M) colony-stimulating activities (CSA), assessed with bioassays and immunoassays. The activities subsided rapidly thereafter. They could be increased by vinblastine, cyclophosphamide, and a sterile inflammatory reaction (s.c. implanted copper rods; Cu-r). Anti-inflammatory indomethacin curtailed the IL-6, but raised the TNF-alpha, G-CSF, and CSA levels in Cu-r mice. Interferon inducer poly-I:C augmented G-CSF, but decreased TNF-alpha levels. Mouse blood cells cultured in chambers expanded their granulocyte/macrophage progenitor population rapidly during the first week of culture; this population being significantly larger on day 3 in perturbed than in unperturbed mice. A marked decline followed in the second week, significantly larger in mice given cytotoxic treatment than in controls. Peritoneal G-CSF and TNF-alpha may explain progenitor and granulocyte growth and development in diffusion chambers during the first week of culture. GM-CSF and IL-3 were apparently without any influence. None of the peritoneal cytokines assayed could explain the population decline during the second week, which was possibly caused by exhaustion of earlier progenitor cells, rather than by intra-chamber feedback mechanisms.

Animals↗
PubMed1976

Fetal hemopoiesis in diffusion chamber cultures. III. The effect of neutropenia.

Proliferation and differentiation of granulocytes, macrophages, and both myeloid committed (CFC) and pluripotent (CFU) stem cells in diffusion chamber (DC) cultures of fetal liver were studied in order to evaluate the role of circulating humoral factors in the control of fetal myelopoiesis. When DC with fetal liver cells were implanted into mice rendered neutropenic by pretreatment with cyclophosphamide, more granulocytes and CFC were produced through day 10 as compared to DC implanted into saline pretreated control hosts. A difference in CFU recovery from fetal liver suspensions grown in DC implanted into neutropenic and control hosts was not seen until day 10. Serum CSF concentrations were increased in neutropenic as compared to control host mice 2 and 3 days after implantation of DC. Levels of serum inhibitors of colony growth showed marked variability but, in general, were similar in both groups. These data provide evidence that fetal CFC and fetal myelopoiesis are influenced by a circulating humoral factor present in neutropenic serum. CSF may be the factor, although the data presented in this paper do not establish this with any certainty.

Agranulocytosis↗
PubMed1976

Unregulated growth of murine leukemic cells and suppression of normal granulocyte growth in diffusion chamber cultures.

The patterns of proliferation of C1498 mouse acute leukemic cells have been studied using the diffusion chamber technique of cell culture. These malignant cells grow to the same maximal cell concentration irrespective of initial cell input. Leukemic cells proliferate equally well with or without the stimulus of prior host irradiation. When cells cultured for several days are diluted to the original input concentration and recultured, they rapidly proliferate back to maximal cell number. All of these findings are in sharp contrast to the behavior of granulocytes from normal mouse marrow grown in the same culture system. Co-culutre of equal numbers of normal marrow cells and leukemic cells results in virtually complete inhibition of normal marrow cells and leukemic cells results in virtually complete inhibition of normal granulocyte growth. These experiments provide a means of studying malignant growth of leukemic cells, as compared to the controlled growth and differentiation of normal granulocytes, and mechanisms by which leukemic cells suppress normal granulocyte development.

Animals↗
PubMed1975

Diffusion chamber culturing of haematopoietic cells: methodological investigations and improvement of the technique.

Mouse bone marrow cells were cultured to determine some basic characteristics of the diffusion chamber (DC) technique. The main findings were as follows: (i) Incorporated 3H-thymidine was conveniently measured after deposition of a cell sample on glass fibre discs followed by methanol washing. Varying the specific activity from approximately 2-approximately 20 Ci/mmole did not affect the relationship between DC cellularity and isotope incorporation. Isotope uptake was similar regardless of whether 3H-thymidine of high or low specific activity had been used. (ii) Higher cell yields were obtained when Millipore or Acropor filters were heat-sealed rather than glued to the plastic rings, when Millipore filters were moistened before DC filling, when we omitted gluing the closing plugs, and when we used an average pore size of 0.22 mum for the DC walls rather than one of 0.10 mum or 0.45 mum. Varying the ring thickness from 2.0-2.5 mm did not impair the cell growth, nor did removal of a softening agent from the plastic rings improve it. (iii) More cells were retrieved from DC carried by young rather than by older mice. Results were not influenced by sex or strain differences between the donor and the host or by the number of implants, whether single or double, within an individual host. (iv) Adding Ficoll to the pronase solution increased the yield of viable CFU-C. (V) Diffusion rate through the DC walls declined with increasing period of culturing, so that i.p. 3H-thymidine is not a flash label for 7-day cultures, for example. The great variability of 3H-thymidine diffusion into i.p. DC was markedly reduced by in vitro exposure to the unopened DC to the isotope. (vi) DC could be incubated in vitro in a medium devoid of protein for at least 6 hours without a fall in 3H-thymidine incorporation rate or CFU-C content, provided that pH was kept constant.

Animals↗
PubMed1985

Potentiality of the murine colony-forming cells detected by an in vivo diffusion chamber culture system.

Culture of a mixture of bone marrow cells with and without T6 chromosome marker in diffusion chambers in mice yielded colonies (CFU-DG) containing cells of a single karyotype, suggesting clonality. Injection of individual CFU-DG colonies into lethally irradiated mice resulted in increased spleen colony formation on day 12 (CFU-S). The possibility of endogenous origin was excluded by demonstrating the presence of T6 marker in both CFU-DG and CFU-S colonies in karyotypically normal hosts. These findings suggest that the cells giving rise to granulocytic colonies in diffusion chambers also can give rise to multipotential hemopoietic cells.

Animals↗