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At least 19 recordsLinked to original sources
PubMed2005

Cell culture block array for immunocytochemical study of protein expression in cultured cells.

Immunocytochemical staining of cultured cells using specific antibodies is a powerful technique to study the expression and subcellular localization of proteins. However, this technique is associated with sample-to-sample variations because samples are handled individually and manually. Cell permeation is needed when intracytoplasmic or nuclear proteins are studied. Storage of cultured cells is difficult, and experiments must be repeated if additional studies are desired later, which introduces more variations. We developed a cell culture block array technique that converts cultured cells into a permanently fixed form identical to tissue sections prepared for pathologic examination. Cells from different cultures can be embedded in a single block. Many identical sections, each containing cells from multiple cultures, may be stained with different antibodies using an automated stainer. As a result, sample-to-sample variation is eliminated. Because cells in these blocks are sectioned by knives, all cellular proteins come into direct contact with antibodies, and cell permeation is not needed. Such blocks can be conveniently stored for years without loss of antigens, providing a constant source for future studies. We demonstrated the utility of this technique by studying the proliferation and neuroendocrine differentiation of prostate cancer-derived LNCaP cells cultured in vitro.

Cell Culture Techniques↗
PubMed2004

A three-dimensional flow control concept for single-cell experiments on a microchip. 1. Cell selection, cell retention, cell culture, cell balancing, and cell scanning.

An ideal microchip for single-cell experiments should be able to allow us to culture cells, to select any desired single cell from a group, to retain the cell for convenient cellular signal detection, and to deliver any buffer or reagent directly to the cell at any time during continual detection and observation. Most importantly, any negative impact on the live cell should be minimized. To accomplish all these functions, we developed a three-dimensional liquid flow control concept and employed special liquid flow fields to manipulate and retain a single yeast cell freely in the chip. A zero-speed point was controlled to retain the cell for three-dimensional cell balancing and cell scanning. A dispersive flow delivered reagents at a high speed to very near the cell and provided them to the cell at a low speed. No force stronger than its gravitational force was exerted on the cell, which could be balanced on different positions on an arc-sloping wall, thus minimizing any negative impact on the cell due to strong liquid flows. Specifically, we demonstrate on-chip single-cell culture, cell wall removal, and reagent delivery. Subsequently, single-cell fluorescence detection was performed, and noise filtering and background correction were applied for data processing.

Cell Culture Techniques↗
PubMed1990

[Cell culture of middle-ear epithelium of the guinea pig--histochemical localization of mitochondrial enzymatic activities in cultured cells].

Primary cell culture system from middle-ear epithelium of the guinea pig was established in defined condition. Mucosal cells were dispersed with enzymatic procedure and over 90% of the cell viability was obtained. Collagen gel and fibronectin coated Thermanox plate were used as culture substrates, and cultured cells on both materials formed confluent epithelial linings. Histochemical localization of succinate dehydrogenase, cytochrome oxidase and adenosine triphosphatase in mitochondria were examined. Cultured ciliated cells and some non-ciliated cells with numerous microvilli showed strong activities of succinate dehydrogenase and cytochrome oxidase. Also in vivo, normal ciliated epithelium near the eustachian tube in the middle-ear cavity of the guinea pig revealed strong mitochondrial metabolic activities. We concluded that this system would be useful for the study of cellular multiplication and differentiation systems of the middle-ear epithelium.

Adenosine Triphosphatases↗
PubMedDate not supplied

New miniaturized hollow-fiber bioreactor for in vivo like cell culture, cell expansion, and production of cell-derived products.

We have developed a miniaturized hollow-fiber bioreactor system for mammalian cell culture with a volume of 1 mL. Cell and medium compartments of the bioreactor are separated by a semipermeable membrane, and oxygenation of the cell compartment is accomplished using an oxygenation membrane. As a result of the geometry of the transparent housing, cells can be observed by microscopy during culture. The leukemic cell lines CCRF-CEM, HL-60, and REH were cultivated up to densities of 3.5 x 10(7)/mL without medium change or manipulation of the cells. As shown using CCRF-CEM cells, growth in the bioreactor was strongly influenced and could be controlled by the medium flow rate. As a consequence, consumption of glucose and generation of lactate varied with flow rate. Depending on the molecular size cutoff of the membranes used, added growth factors such as GM-CSF, as well as factors secreted from the cells, are retained in the cell compartment for up to 1 week. This new miniaturized hollow-fiber bioreactor offers advantages in tissue engineering by continuous nutrient supply for cells in high density, retention of added or autocrine produced factors, and undisturbed long-term culture in a closed system.

Animals↗
PubMed2005

Poly-L-lactide surfaces subjected to long-term cell cultures: cell proliferation and polymer degradation.

Short term cell cultures are usually grown in contact with biomaterials to assess cytocompatibility. Depending on the rate of material degradation or corrosion, the time of culture can be a key-point in the method which, if too short, may not show any effect of the released material on the cells. A long term culture was therefore carried out with L929 fibroblast cells in contact with PLLA/PDLA samples for up to eight months. The degradation was measured in terms of shear-strength properties, intrinsic viscosity of the material and its cristallinity. The effect of the material on the cells was evaluated by measuring the growth rate of the cells. A significant decrease in the shear strength of the material was measured after three months. The rate of modification of the intrinsic viscosity was regular and decreased progressively throughout the culture period. Differential scanning calorimetry showed that the samples were initially essentially amorphous and that contact with the cell culture and its medium did not change its crystallinity level. The growth rate of the cells was not modified by the presence of the material when compared to the control. This study showed this material to be cytocompatible for a long period of time, even after detection of modifications of its physico-chemical properties.

Animals↗
PubMed1983

Assay of anticancer drugs in tissue culture: cell cultures of biopsies from human astrocytoma.

A method has been developed for measuring the drug sensitivity of human gliomas in short-term culture, using scintillation counting or autofluorography. Cell cultures prepared from malignant astrocytomas were treated with anticancer drugs whilst in exponential growth in microtitration plates. After drug treatment and a recovery period, residual viability was measured by [3H] leucine incorporation followed by scintillation counting or by [35S] methionine incorporation and autofluorography in situ. In 5 glioma cell lines tested against 6 drugs, the microtitration method correlated well with monolayer cloning. Although replicate samples of the same tumour showed little variation in chemosensitivity, there was marked variation between the chemosensitivities of cultures derived from the tumours of different patients. However, as variability between replicates was apparent during drug exposure or shortly after, it is important to allow the assay to run as long as possible after drug removal. It is hoped that this assay may provide the basis of a method for the prediction of in vivo chemosensitivity or the screening of potential chemotherapeutic drugs.

Antineoplastic Agents↗
PubMed1990

[Antibodies to cell culture cells as an interference factor in chlamydia serology].

Antibodies against Chlamydia trachomatis were detected with the immunoperoxidase assay (IPA) in 32 patients suffering from rheumatic diseases. In this test the cells infected with Chlamydia trachomatis serotype L2 were the antigen. In addition all sera were tested for anticellular antibodies, using non-infected cells as antigen. 13 out of these 32 IPA-positive patients (41%) were also positive with respect to anticellular antibodies.

Antibodies↗
PubMed1980

Mode of subacute sclerosing panencephalitis (SSPE) virus infection in tissue culture cells. II. Cell-free viruses in cell cultures infected with Kitaken-1 and Biken strains of SSPE virus.

Cell-free infectious viruses were successfully recovered by the aid of freezing and thawing from cultures infected with the Kitaken-1 and Biken strains of subacute sclerosing panencephalitis (SSPE) virus. Our results including those in a previous report which dealt with the Niigata-1 strain of SSPE virus show that cell-free viruses can be detected from all of the SSPE virus-carrying cultures established in Japan. It was also found that cell-free infectious viruses can be recovered efficiently by dispersing the virus-carrying cultures with EDTA. The inclusion of trypsin in the EDTA solution, however, caused a poor recovery of the infectious viruses. Infection of cells with the cell-free viruses readily established the virus-carrying cultures that have characteristics comparable to those of their original cultures. The culture infected with the Kitaken-1 strain produced infectious viruses in about ten times the amount of the other two infected cultures. The buoyant densities of the cell-free infectious viruses were almost the same among the three strains, the values being 1.120 to 1.132, but significantly less than that of 1.164 of measles virus. The low density can be ascribed to one of the characteristics of these SSPE viruses.

Cell Line↗
PubMed1989

Evaluation of techniques to demonstrate foot-and-mouth disease virus in bovine tongue epithelium: comparison of the sensitivity of cattle, mice, primary cell cultures, cryopreserved cell cultures and established cell lines.

Tongue epithelia infected with each of the 7 serotypes of foot-and-mouth disease virus (FMDV) were used to evaluate in vivo and in vitro systems for the detection of FMDV. Cattle inoculated by the intradermal route in the tongue (IDL) and suckling mice inoculated intraperitoneally were compared for susceptibility to FMDV with freshly prepared bovine thyroid cell cultures; cultures from cryopreserved bovine thyroid, bone marrow, mammary gland, myocardium, tongue, ovary and kidney cells; cultures from cryopreserved embryonic ovine kidney, newborn ovine kidney, ovine testicle, bone marrow, and chloroid plexus cells; and the continuous porcine kidney cell lines MVPK-1 and S6. The mean titers determined for each serotype in each system were statistically compared. The FMDV titers obtained in freshly prepared bovine thyroid cell cultures and by cattle IDL inoculation were the highest and were statistically indistinguishable. The titers obtained by suckling mouse inoculation were significantly lower than the titers obtained in thyroid cultures for serotypes A, C, Asia 1, and SAT 3. The cattle IDL assay was significantly more sensitive than the mouse assay for serotype A. The cell cultures from the cryopreserved newborn ovine kidney and embryonic ovine kidney were significantly less susceptible to serotype Asia 1 when compared with the fresh bovine thyroid cultures, but not significantly different when compared with the cattle assay for all serotypes. Cryopreservation of bovine thyroid cells directly after trypsinization resulted in the loss of susceptibility to FMDV serotype SAT 2. The other cryopreserved cell culture systems exhibited no or minimal susceptibility to all 7 serotypes, or exhibited considerable inconsistency. The established cell lines MVPK-1 and S6 were not susceptible to serotype A, and were less sensitive to serotype C than other culture systems. Quality control of cell cultures used to evaluate field specimens for FMDV was critical. The cell cultures of cryopreserved ovine kidney cells provided the most practical diagnostic system.

Animals↗