scholarly journals Simple Method for Establishing Primary Leporidae Skin Fibroblast Cultures

Cells ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 2100
Author(s):  
Fábio A. Abade dos Santos ◽  
C. L. Carvalho ◽  
Isabel Almeida ◽  
Teresa Fagulha ◽  
Fernanda Rammos ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Enzymatic Digestion ◽  
Primary Fibroblast ◽  
Simple Method ◽  
Cheap Method ◽  
Immortalized Cell ◽  
The Many

Commercial hare and rabbit immortalized cell lines are extremely limited regarding the many species within the lagomorpha order. To overcome this limitation, researchers and technicians must establish primary cell cultures derived from biopsies or embryos. Among all cell types, fibroblasts are plastic and resilient cells, highly convenient for clinical and fundamental research but also for diagnosis, particularly for viral isolation. Here, we describe a fast and cheap method to produce primary fibroblast cell cultures from leporid species, using dispase II, a protease that allows dermal–epidermal separation, followed by a simple enzymatic digestion with trypsin. This method allows for the establishment of an in vitro cell culture system with an excellent viability yield and purity level higher than 85% and enables the maintenance and even immortalization of leporid fibroblastic cells derived from tissues already differentiated.

Phenotypic plasticity in the intercalated cell: the hensin pathway

1998 ◽  
Vol 275 (2) ◽  
pp. F183-F190 ◽  
Author(s):  
Qais Al-Awqati ◽  
S. Vijayakumar ◽  
C. Hikita ◽  
J. Chen ◽  
J. Takito
Keyword(s):  
Collecting Duct ◽  
Basolateral Membrane ◽  
Cell Types ◽  
Band 3 ◽  
Cytoskeletal Proteins ◽  
Β Cell ◽  
Biochemical Basis ◽  
Intercalated Cell ◽  
Immortalized Cell

The collecting duct of the renal tubule contains two cell types, one of which, the intercalated cell, is responsible for acidification and alkalinization of urine. These cells exist in a multiplicity of morphological forms, with two extreme types, α and β. The former acidifies the urine by an apical proton-translocating ATPase and a basolateral Cl/HCO3 exchanger, which is an alternately spliced form of band 3. This kidney form of band 3, kAE1, is present in the apical membrane of the β-cell, which has the H+-ATPase on the basolateral membrane. We had suggested previously that metabolic acidosis leads to conversion of β-types to α-types. To study the biochemical basis of this plasticity, we used an immortalized cell line of the β-cell and showed that these cells convert to the α-phenotype when plated at superconfluent density. At high density these cells localize a new protein, which we term “hensin,” to the extracellular matrix, and hensin acts as a molecular switch capable of changing the phenotype of these cells in vitro. Hensin induces new cytoskeletal proteins, makes the cells assume a more columnar shape and retargets kAE1 and the H+-ATPase. These recent studies suggest that the conversion of β- to α-cells, at least in vitro, bears many of the hallmarks of terminal differentiation.

PRIMARY FIBROBLAST CELL CULTURES FROM TRANSBRONCHIAL BIOPSIES OF LUNG TRANSPLANT RECIPIENTS1

2001 ◽  
Vol 71 (2) ◽  
pp. 337-339 ◽  
Author(s):  
Michael Tamm ◽  
Michael Roth ◽  
Monique Malouf ◽  
Prashant Chhajed ◽  
Peter Johnson ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Lung Transplant ◽  
Fibroblast Cell ◽  
Primary Fibroblast

Amphiregulin Induces Proliferative Activities in Osseous Dysplasia

2009 ◽  
Vol 88 (6) ◽  
pp. 563-568 ◽  
Author(s):  
H. Shigeishi ◽  
S. Yamaguchi ◽  
K. Mizuta ◽  
K. Nakakuki ◽  
S. Fujimoto ◽  
...  
Keyword(s):  
Cellular Differentiation ◽  
Expression Patterns ◽  
Cell Types ◽  
Gingival Fibroblasts ◽  
Osseous Dysplasia ◽  
The Difference ◽  
Immortalized Cell ◽  
Differentiation And Proliferation ◽  
Characteristic Growth

Human osseous dysplasia (OD) is a benign fibro-osseous neoplasm of periodontal ligament origin in which normal bone is replaced with fibrous connective tissue containing abnormal bone or cementum. However, cellular differentiation and proliferation in OD have not been fully elucidated. In vitro culture systems have distinct advantages for analytical studies. Therefore, we established immortalized cell lines (OD-1) from OD lesions of the jaw from an individual with gnathodiaphyseal dysplasia (GDD). We hypothesized that OD-1 had a characteristic growth mechanism different from that of mineralized-associated cells such as osteoblasts. To clarify the difference of gene expression patterns between OD-1 and osteoblasts, we compared the profiles of genes expressed in the 2 cell types by microarray analysis. We identified amphiregulin to be highly expressed in OD-1 compared with osteoblasts and gingival fibroblasts. OD-1 showed proliferative activities regulated in an autocrine manner by amphiregulin, and amphiregulin may play a significant role in the proliferation of OD.

scholarly journals Attachment and invasion ofToxoplasma gondiiandNeospora caninumto epithelial and fibroblast cell linesin vitro

Parasitology ◽  
2005 ◽  
Vol 131 (5) ◽  
pp. 583-590 ◽  
Author(s):  
YING LEI ◽  
M. DAVEY ◽  
J. T. ELLIS
Keyword(s):  
Cell Lines ◽  
Neospora Caninum ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Vero Cells ◽  
Host Cells ◽  
Trypsin Treatment ◽  
Epithelial Cell Lines ◽  
Pre Treatment

Attachment and invasion ofToxoplasma gondiiandNeospora caninumto a cat and a dog fibroblast cell line and 2 epithelial cell lines (a cat kidney and Vero) were comparedin vitrousing fluorescence antibody methodology. In addition, trypsin treatment of tachyzoites was used to determine whether protein molecules were essential to the process of invasion. The results show that bothT. gondiiandN. caninuminvaded all 4 cell lines, and that pre-treatment ofT. gondiitachyzoites with trypsin caused an increase in the ability of the parasite to invade these host cells. FurthermoreT. gondii, in comparison toN. caninum, invaded all 4 cell lines at greater levels. The results here support the conclusion that bothT. gondiiandN. caninumhave the ability to invade a variety of cell types including both dog and cat cells, and questions the utility of Vero cells as an appropriate host cell forin vitrostudies on the biology of these taxa.

scholarly journals Endothelial injury in scleroderma.

1979 ◽  
Vol 149 (6) ◽  
pp. 1326-1335 ◽  
Author(s):  
M B Kahaleh ◽  
G K Sherer ◽  
E C LeRoy
Keyword(s):  
Endothelial Cells ◽  
Present Report ◽  
Gel Filtration ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Endothelial Damage ◽  
Vascular Lesions ◽  
Vascular Changes ◽  
Heat Stable

Functional and structural vascular lesions have been observed in the organs involved in scleroderma. The etiology of these vascular changes is poorly understood. The ability to isolate, characterize, and maintain endothelial cells in vitro provides a target cell population to study endothelial damage in scleroderma. The present report describes the effect of scleroderma serum on endothelial, smooth muscle, and fibroblast cell types. Sera from patients with scleroderma (31/52) and Raynaud's syndrome (11/19) contain cytotoxic activity, specific for endothelial cells, which is nondialyzable, heat-stable, and elutes with albumin on gel-filtration chromatography.

scholarly journals Cell proliferation induced by modified cationic dextran

2018 ◽  
Vol 14 (4) ◽  
Author(s):  
Kamil Kamiński ◽  
Krystyna Stalińska ◽  
Anna Niziołek ◽  
Maria Wróbel ◽  
Maria Nowakowska ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Lines ◽  
Positive Impact ◽  
Cell Aggregation ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Cationic Modification ◽  
Diphenyl Tetrazolium Bromide ◽  
The Impact

Abstract The interaction between oppositely charged membranes and polycations causes cell aggregation, loss of membrane fluidity, and membrane degeneration and may cause an increase of its permeability. Unfortunately, the interaction is the reason why the use of polycations in medicine is severely limited. Therefore, in this paper, we share our observations related to the action of 40-kDa dextran modified using glycidyltrimethylammonium chloride, resulting in increased fibroblast cell proliferation. Using viability and proliferation tests [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, crystal violet, 3H-thymidine incorporation], we have observed that cationic dextran derivatives exert a positive impact on nonepithelial cell proliferation in vitro. This phenomenon has been noted for human and mouse fibroblasts and several other nonepithelial cell lines. However, the effect seems to be most pronounced for fibroblast cell lines. The presented studies allow to examine the impact of the polymer structure and the methods of its cationic modification on this newly observed phenomenon. The observation is unique because positively charged macromolecules usually exhibit high toxicity in all cell types in vitro.

scholarly journals Lipid-Derived Mediators in Endogenous Anti-Inflammation and Resolution: Lipoxins and Aspirin-Triggered 15-epi-Lipoxins

2002 ◽  
Vol 2 ◽  
pp. 169-204 ◽  
Author(s):  
Charles N. Serhan ◽  
Nan Chiang
Keyword(s):  
Cell Types ◽  
Cellular Systems ◽  
Eicosatetraenoic Acid ◽  
Small Peptides ◽  
On Line ◽  
The Many ◽  
G Protein Coupled ◽  
Physiologic Responses

It is well appreciated that lipid-derived mediators play key roles in inflammation and many other physiologic responses where multicellular processes are involved. Among them, lipoxins (LX) and aspirin-triggered LX (ATL) evoke actions of interest in a range of physiologic and pathophysiologic processes, and these two series have emerged as founding members of the first class of lipid/chemical mediators “switched on” in the resolution phase of an inflammatory reaction. These unique compounds possess a trihydroxytetraene structure and are both structurally and functionally distinct among the many groups of lipid-derived bioactive mediators. LXA4 and 15-epi-LXA4(a member of the ATL series) display leukocyte-selective actions that enable them to serve as endogenous “stop signals” in multicellular events in that they modulate adherence, transmigration, and chemotaxis. Both LXA4and 15-epi-LXA4elicit these responses via a G protein-coupled receptor (GPCR), termed ALXR, identified in human and murine tissues. Among eicosanoids, ALXR is stereoselective for LXA4(5S,6R,15S-trihydroxy-7,9,13-trans-11-cis-eicosatetraenoic acid). Its aspirin-triggered 15R epimer (15-epi-LXA4) and their bioactive stable analogs act in the subnanomolar to nanomolar range in human cellular systems and murine models of acute inflammation and reperfusion. ALXR also has the ability to interact with a wide panel of small peptides that give different signaling responses in vitro than LXA4or its analogs, suggesting that ALXR is capable of serving as a multirecognition receptor in immune responses. Characterization of ALXR and development of metabolically stable LX and ATL analogs that are mimetics rapidly advanced our appreciation of the mechanism of LX actions and the potential utility of these counter-regulatory biocircuits in the quest to control local inflammatory events. In this on-line update, LX and ATL biosynthesis and the LXA4specific receptor, termed ALXR, are reviewed with a focus on their roles in inflammation and resolution with respect to pharmacology, molecular biology, and signal transduction in several cell types and animal models investigated thus far.

scholarly journals THE RATE OF DIVISION OF ANTIBODY-FORMING CELLS DURING THE EARLY PRIMARY IMMUNE RESPONSE

1968 ◽  
Vol 127 (5) ◽  
pp. 983-1002 ◽  
Author(s):  
Donald A. Rowley ◽  
Frank W. Fitch ◽  
Donald E. Mosier ◽  
Susan Solliday ◽  
Lionel W. Coppleson ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Drug Treatment ◽  
Cell Cultures ◽  
Cell Types ◽  
Primary Immune Response ◽  
Spleen Cells ◽  
Sheep Erythrocytes ◽  
Cycle Times ◽  
Blocking Agents

Mitotic blocking agents, colchicine or Velban, were used to estimate cycle times of spleen cells which release hemolysin for sheep erythrocytes (plaque-forming cells). The cells were obtained either from rats immunized with sheep erythrocytes or from cultures of mouse spleen cells immunized in vitro with the same antigen. 2, 3, or 4 days after immunization, animals or cell cultures were treated with mitotic blocking agents for periods of time ranging from 2.5 to 7 hr; plaque-forming cells were then enumerated. Decreased numbers of plaque-forming cells were found after such treatment. The extent of reduction was a function of duration of the drug treatment and the method of immunization, but was independent of the time after immunization. The evidence presented is consistent with premises that: (a) plaque-forming cells in mitosis do not release sufficient antibody to be detected, (b) mitotic blocking agents, by arresting plaque-forming cells in metaphase, prevent not only detection of these cells but also the increase in number of plaque-forming cells which would have resulted from cell division, (c) mitotic blocking agents do not affect release of antibody by cells in interphase. Cell cycle times, based on the extent of reduction of plaque-forming cells per unit time of drug treatment, were estimated using a mathematical model appropriate for an exponentially increasing population of cells. Cell cycle times estimated using the mitotic blocking agents agreed well with cell doubling times calculated from the increase in plaque-forming cells occurring 1–4 days after immunization. Increased responses produced by higher antigen doses or treatment of immunized animals with an adjuvant resulted from an increased rate of division of responding cells and their progeny. The results are consistent with a cell selection theory of antibody formation. Antigenic stimulation causes relatively few cells to proliferate and to synthesize antibody; apparently the magnitude of the response is dependent primarily on the rate of division of responding cells. It is suggested on the basis of observations of in vitro-immunized cell cultures that the rate of division of responding cells may be dependent on the rate of interaction between two cell types, both of which are essential for the in vitro plaque-forming cell response.

The production and effect of interferon on influenza virus growth in chick embryo lung epithelial and fibroblast cell cultures

1969 ◽  
Vol 15 (3) ◽  
pp. 273-277 ◽  
Author(s):  
Sunidhkumar S. Gandhi ◽  
Robert B. Stewart
Keyword(s):  
Epithelial Cells ◽  
Chick Embryo ◽  
Cell Cultures ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Fibroblastic Cell ◽  
Virus Growth ◽  
Lung Epithelial ◽  
Fibroblastic Cells ◽  
Virus Strains

Cultures of fibroblastic cells prepared from chick embryo lung infected with low multiplicities of influenza type A virus strains were found to produce more interferon than did cultures of epithelial cells prepared from the same organ. Fibroblastic cell cultures were also found to be more sensitive to the action of interferon than were epithelial cells with respect to the levels of infectious virus produced and the duration of interferon action. Cultures of the two cell types treated with interferon did not differ with respect to the number of cells involved in virus synthesis.

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