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At least 19 recordsLinked to original sources
PubMed1991

Biocompatibility evaluation of glass ionomer cement using cell culture techniques.

Cell culture techniques have been employed to carry out a preliminary investigation into the biocompatibility of a glass ionomer cement (GIC). A modification of the commonly used Agar Overlay test and a rapid, colorimetric assay based on the reduction of tetrazolium salt were used. The GIC is shown to leach a cytotoxic agent, possibly fluoride. This agent is effectively removed by an extraction procedure, and re-examination of the GIC demonstrates its cytocompatibility.

Agar↗
PubMed1980

Studies of renal cell function using cell culture techniques.

Cell culture, a powerful tool for the study of cell biology, offers advantages for the study of renal cell function. Epithelial cells derived from a variety of organs, including the kidney, form oriented epithelial sheets in culture that have many structural characteristics (microvilli, tight junctions) of epithelia in situ. There is evidence of transepithelial transport of salt and water by cells of two lines (MDCK and LLC-PK1) derived from mammalian kidney. LLC-PK1 cells may also manifest the glucose transport system of the proximal tubule. Cells of both lines have adenylate cyclase activity sensitive to hormones. Two lines of cells derived from toad urinary bladder form epithelia with a high transepithelial resistance and transport sodium actively from apical to basolateral surface. The rate of sodium transport in both lines is stimulated by cyclic AMP and by aldosterone. There are important differences in the characteristics of the response of the two lines to aldosterone as well as in their sensitivity to inhibition of sodium transport by amiloride. These differences may lead to new insights regarding the molecular events in the response to aldosterone and in the inhibitory action of amiloride. Cultures of kidney cells have also been used effectively to study the biosynthesis of the hormonal derivative of vitamin D and to study prostaglandin production. In addition, cell culture is ideally suited for study of the developmental biology of the kidney.

Aldosterone↗
PubMedDate not supplied

Cerebrospinal fluid cytology in patients with brain tumors; a simple method using the cell culture technique.

A simplified cytologic method using the cell culture technique was employed in 71 cases with brain tumor. Neoplasitc cells were demonstrated in 17 cases (24%), that is in 11 out of 30 cases of glioma, four out of ten cases of metastatic brain tumor, and two out of 17 cases of meningioma. None of 14 other miscellaneous tumors proved positive. Identification of glioma cells could be easily made because they usually showed characteristic morphology and good proliferation in vitro. However, for other types of tumor, conventional methods were considered to be superior to the culture method because their exfoliated cells usually underwent rapid degeneration without showing characteristic morphology during the culture. Some of the illustrative cases were presented.

Adolescent↗
PubMed2001

[Rat keratinocyte primary cultures based on conductive polypyrrole primary cell culture technique].

Orthogonal experimental design has been applied to study the suitable cell culture medium and condition of conductive polypyrrole on rat keratinocyte cell cultures. The results showed that one thousand milliliters of complete minimum essential medium(CMEM) which contained 900 ml minimum essential medium(MEM), 10% fetal bovine serum(FBS), 1% NEAA, 6 ml insulin and 10 mg hydrocortisone was fit for keratinocyte cell culture. And the conductive polypyrrole obtained with 10 mA current density for 100 second switching times, given 100 mV electrical stimulation voltage for 2 hours, would stimulate the growth of rat keratinocyte cell in vitro cultures. The cell density under culture conditions described above was 1.30 times as high as that under routine culture conditions.

Animals↗
PubMed2002

Biotechnology apprenticeship for secondary-level students: teaching advanced cell culture techniques for research.

The purpose of this article is to discuss small-group apprenticeships (SGAs) as a method to instruct cell culture techniques to high school participants. The study aimed to teach cell culture practices and to introduce advanced imaging techniques to solve various biomedical engineering problems. Participants designed and completed experiments using both flow cytometry and laser scanning cytometry during the 1-month summer apprenticeship. In addition to effectively and efficiently teaching cell biology laboratory techniques, this course design provided an opportunity for research training, career exploration, and mentoring. Students participated in active research projects, working with a skilled interdisciplinary team of researchers in a large research institution with access to state-of-the-art instrumentation. The instructors, composed of graduate students, laboratory managers, and principal investigators, worked well together to present a real and worthwhile research experience. The students enjoyed learning cell culture techniques while contributing to active research projects. The institution's researchers were equally enthusiastic to instruct and serve as mentors. In this article, we clarify and illuminate the value of small-group laboratory apprenticeships to the institution and the students by presenting the results and experiences of seven middle and high school participants and their instructors.

Animals↗
PubMed1985

Alterations in the phosphoprotein composition of tumor cell clones induced by cell culture techniques used routinely in assessing metastatic behavior in vivo.

To investigate whether the cell dispersion techniques commonly employed to harvest monolayer cultures of tumor cells for injection into experimental animals might induce alterations in cellular biochemistry, we compared the phosphoprotein profiles of 6 B16 melanoma clones of distinct metastatic potential using 2-D gel electrophoresis after growth in monolayer culture, after suspension by treatment, with trypsin/EDTA and after injection of suspended cells into syngeneic mouse plasma. Trypsin/EDTA treatment and subsequent exposure to syngeneic mouse plasma induced significant alterations in phosphoprotein composition in all clones. Most alterations were quantitative, involving either enhanced or diminished expression of specific phosphoproteins, but qualitative changes involving expression of novel phosphoproteins were also observed. None of the changes in phosphoprotein composition correlated with metastatic potential. The principle alteration induced in all clones by trypsin/EDTA involved enhanced phosphorylation of an NP-40-soluble component with a molecular weight of 79,000 and an isoelectric point of 6.3 [pp79 (6.3)]. This determinant was detected in extracts of B16 monolayer cultures but its level of phosphorylation was enhanced significantly by trypsin/EDTA treatment and by exposure of the harvested cells to syngeneic mouse plasma. These data indicate that procedures commonly employed to harvest tumor cells for assay of tumorigenic and metastatic potential may provide extensive alterations in phosphoprotein composition and that biochemical investigations of tumor cells grown in monolayer culture may not accurately reflect the metabolic status of the same cells immediately prior to and following i.v. injection into experimental animals.

Animals↗
PubMed1990

Virological applications of the grid-cell-culture technique.

Whole mounts of intact virus-infected cells have been used for several decades to examine virus-cell relationships and virus structure. The general concept of studying virus structure in association with the host cell has recently been expanded to reveal interactions between viruses and the cytoskeleton. The procedure permits utilization of immuno-gold protocols using both the transmission and scanning electron microscopes. The grid-cell-culture technique is reviewed to explain how it can be exploited to provide valuable information about virus structure and replication in both diagnostic and research laboratories. The use of the technique at the research level is discussed using bluetongue virus as a model. The procedure can provide basic structural information about intact virions and additional data on the intracellular location of viruses and virus-specific structures and about the mode of virus release from infected cells. Application of immunoelectron microscopy reveals information on the protein composition of not only released virus particles but also cell surface and cytoskeletal-associated viruses and virus-specific structures. Collectively, this simple and physically gentle technique has provided information which would otherwise be difficult to obtain.

Bluetongue virus↗
PubMed1991

[In vitro toxicity evaluation of 12 kinds of dental materials using a modified cell culture technique].

In view of the fact that up to date the various cell toxicity tests could not come to an identical result and compare with each other. This paper, referring to the covering documents of F.D.I. and I.S.O., summing up the principles and characteristics of various cell culture techniques, establishes a modified cell toxicity test. This method, with more respects of wide observations and tangible results, is easy to grasp, well to repeat and can be reserved for checking the results, meanwhile, it can be avoided the contradiction caused by the different experiments on one kind of material. Therefore, it is very worthy of appraising the biological safety of the materials. The study of 12 kinds of dental materials expressed that the toxicity order of copper alloy was mean value of 0 (Jiao Tong University), copper alloy tridium was mean value of 3 (U.S.A.), copper alloy T was mean value of 2 (Jiao Tong University) and the others were mean value of 0-1. This paper also discussed several problems with common concerns in studying the cell toxicity of the biomaterials.

Animals↗
PubMed1986

An established cell line from mouse kidney medullary thick ascending limb. I. Cell culture techniques, morphology, and antigenic expression.

The effective use of cultured cells as model systems for investigating the differentiation and regulation of transport processes in renal tubular epithelial cells depends on the availability of functional long-term cell lines derived from specific nephron segments. Conventional culture procedures that treat cells as proliferating microorganisms possess several inherent limitations that could contribute to phenotypic instability and limited proliferative capacity in vitro. In this study, culture techniques were adopted that avoid exposure of cells to proteolytic enzymes, maintain intercellular contacts, and allow cells to remain continually adherent to a collagen gel substratum. This methodology resulted in the development of a continuous epithelial cell line from identified, microdissected segments of the mouse kidney medullary thick ascending limb.

Animals↗
PubMed1978

The r.b.e. of different-energy neutrons as determined by human bone-marrow cell-culture techniques.

The effect of X-rays and different-energy neutrons on human bone-marrow cells was studied using two different cell-culture techniques--diffusion chamber (DC) growth and colony formation in vitro (CFU-C). Based on the survival of proliferative granulocytes in DC on day 13, the D0 value was 80 rad with X-rays, and 117 rad as measured by the CFU-C assay. The D0 values for neutrons depended on the radiation source and the energy level. The r.b.e. values, which dropped with increasing energy levels of mono-energetic neutrons, were (i) 0.44 MeV; DC 3.7, CFU-C 4.1; (ii) 6 MeV; DC 1.8, CFU-C 2.0; (iii) 15 MeV; DC 1.6, CFU-C 1.6; (iv) fission neutrons; DC 2.6, CFU-C 2.4.

Bone Marrow↗
PubMed2006

Biocompatibility of furcal perforation repair material using cell culture technique: Ketac Molar versus ProRoot MTA.

OBJECTIVE: The objective of this study was to evaluate the cytotoxicity of furcal perforation repair materials, GI and MTA, using cell culture technique. STUDY DESIGN: The extract of ProRoot MTA and Ketac Molar were treated on PDL cells in a 96-well tissue-culture plate. Cell proliferation after an incubation period of 3 days was determined by using MTT assay. RESULTS: The growth of cultured human periodontal fibroblast cells were suppressed by both perforation repair materials. The percent of cell viability in the Ketac Molar group was lower than in the ProRoot MTA group (P = .000). CONCLUSIONS: Although Ketac Molar has the advantage of adhering to dentine, it is more cytotoxic to the PDL cells than MTA. In selecting the perforation repair material, it is recommended not only to consider the sealing ability of the material with dentine but also the biocompatibility of material to the underlying tissue.

Aluminum Compounds↗
PubMed1982

Evaluation of the biologic effects of dental materials using a new cell culture technique.

The biological effects of seven dental cements--including zinc oxide-eugenol, EBA, silicate, zinc phosphate, silico-phosphate, polycarboxylate, and glass ionomer--and a composite resin were evaluated by a new cell culture technique. The agar overlay containing serum was proposed as a dentin model. The effects of thickness and serum concentration of the agar overlay on cell growth were studied and compared to in vivo findings. Some correlative phenomena between the cell culture results and in vivo data are presented.

Cells, Cultured↗
PubMed1977

Evaluation of biologic effects of dental materials using four different cell culture techniques.

The cytotoxicity of fresh and 1-day-old silicate cement, composite restorative material and zinc oxide-eugenol cement (ZOE) was tested using human epithelial cells (NCTC 2544) in four different cell culture systems: (1) 51Cr-release from prelabeled cells after incubation for 4 and 24 h in the presence of the materials. (2) Implanting the materials on an agar everlay and visualizing any cytotoxic effects after 24 h by neutral red vital stain. (3) Cell growth during 5 d in the presence of the materials. (4) Colony-forming ability after exposure of the cells for 30 min to medium previously incubated with the materials for 24 h. Freshly prepared and 1-day-old ZOE exhibited a prominent cytotoxic effect in all four systems. A less marked effect was found for the composite material in systems 2, 3, and 4, while silicate cement appeared to be the least toxic material in these three systems. Neither silicate cement nor composite showed any cytotoxic effect in system 1 based on 51Cr-release. It is concluded that the effects obtained by the cell culture techniques did not mimic the reactions obtained when the materials are tested under conditions which reflect their clinical use.

Cell Division↗
PubMed1987

The grid-cell-culture technique: the direct examination of virus-infected cells and progeny viruses.

We describe a method for the structural analysis and identification of viruses, without purification or concentration steps which could alter virus morphology. Virus-infected cells grown on carbon-Parlodion-coated electron microscope grids release large numbers of progeny viruses which adsorb to the surface of the grid and are revealed by negative staining. The technique is rapid, sensitive and can be used at three levels. Negative staining of whole cell preparations revealed both extracellular and intracellular viruses or nucleocapsids beneath the plasma membrane; non-ionic detergent extraction of cells infected with certain viruses reveals cytoskeleton-associated, virus-specific structures normally only observed after thin sectioning; cultures prepared by either procedure are suitable for colloidal gold immunological studies. Extracellular and cytoskeletal-associated viruses were heavily and specifically labelled with gold. The results indicate that the technique may be used to rapidly identify unknown viruses on the basis of size, topography, morphology and mode of maturation from the infected cell, as well as the presence of characteristic intracellular cytoskeletal-associated structures. The technique also has potential use in the sero-grouping and sero-typing of viruses with specific monoclonal antibodies.

Animals↗
PubMed1988

New and sensitive standard cell culture technique for the detection of cytomegalovirus in clinical specimens.

Conventional tube cell culture has been recognised as the most sensitive technique available for human cytomegalovirus (HCMV) detection. Low-speed centrifugation of specimen inocula onto cell culture monolayers has been shown to increase the efficiency of infection with the AD 169 strain of HCMV. Therefore a centrifugal force of 900g for 1 hour at 37 degrees C was used to enhance the detection of HCMV cytopathic effect (CPE) in shell vials that contained circular coverslips with a monolayer of human embryonic lung (HEL) fibroblasts. Of 195 specimens, HCMV CPE was detected in 18 specimens (9.02%) on shell vial culture assay, whereas conventional tube cell culture was positive in only 13 specimens (6.6%). The shell vial culture assay was significantly more sensitive (P less than 0.05). Furthermore the development of the cytopathic effect on shell vial culture assay was significantly earlier (P less than 0.01) and more extensive. Urine samples were sonicated and the results obtained with immunofluorescence using human immune serum demonstrated that sonication increased both the intensity of fluorescence and number of fluorescent foci of HCMV-infected cells and also decreased the non-specific fluorescence of the background.

Cells, Cultured↗
PubMed1996

Tissue engineering and autologous transplant formation: practical approaches with resorbable biomaterials and new cell culture techniques.

The engineering of living tissues in vivo requires new concepts in cell culture technology. In contrast to conventional cell cultures, the development of tissues depends on a three-dimensional arrangement of cells and the formation or synthesis of an appropriate extracellular matrix. Special emphasis is given to the major role of the extracellular matrix and cell differentiation in an artificial tissue. New technical approaches of in vitro tissue engineering are compared to the natural development of tissues in vivo. Current methods using resorbable biomaterials, tissue encapsulation and perfusion culture are discussed. Major consideration is given to scaffold structures of biomaterials that define a three-dimensional shape of a tissue or guide matrix formation. The different goals of tissue engineering such as in vitro models and transplant production are taken into account in the described techniques. Practical concepts comprising cell multiplication and differentiation in subsequent steps for future clinical applications are outlined.

Animals↗