Conditioned medium mediated immune protection in In Vitro model of endogenous neurotoxin N-methyl-salsolinol in an astrocyte dependent manner

2015 ◽  
pp. 217-220
Keyword(s):  
Conditioned Medium ◽  
In Vitro Model ◽  
Dependent Manner ◽  
Immune Protection ◽  
Vitro Model

In vitro study of placental trophoblast calcium uptake using JEG-3 human choriocarcinoma cells

1991 ◽  
Vol 98 (3) ◽  
pp. 333-342
Author(s):  
R.S. Tuan ◽  
C.J. Moore ◽  
J.W. Brittingham ◽  
J.J. Kirwin ◽  
R.E. Akins ◽  
...  
Keyword(s):  
Vitamin D3 ◽  
Calcium Uptake ◽  
In Vitro Model ◽  
Dependent Manner ◽  
Choriocarcinoma Cells ◽  
Vitro Model ◽  
Human Choriocarcinoma Cells ◽  
Functional Components ◽  
1,25 Dihydroxy Vitamin D3

During human fetal development, placental syncytiotrophoblasts actively transport calcium from the maternal to the fetal circulation. Two functional components, a cytosolic Ca2(+)-binding protein (CaBP) and a Ca2(+)-ATPase have been identified in the syncytiotrophoblasts of the chorionic villi. We report here the calcium uptake properties of a human choriocarcinoma cell line, JEG-3, which was used as an in vitro model cell system for the syncytiotrophoblasts. In culture, JEG-3 proliferated as large syncytial aggregates expressing typical syncytiotrophoblast markers. 45Ca uptake by JEG-3 was a substrate- and temperature-dependent, membrane-mediated active process that exhibited linear kinetics for up to 7 min. Both the CaBP and the Ca2(+)-ATPase were expressed by JEG-3, on the basis of biochemical, histochemical, immunochemical and or mRNA assays. Immunohistochemistry and in situ hybridization revealed that JEG-3 cells were heterogeneous with respect to the expression of the CaBP. The Ca2(+)-ATPase activity of JEG-3 was similar to the placental enzyme in terms of sensitivity to specific inhibitors, and was detected histochemically along the cell membrane. Fura-2 Ca2+ imaging revealed that calcium uptake by JEG-3 was not accompanied by a concomitant increase in cytosolic [Ca2+], suggesting a specific Ca2+ sequestration mechanism. The involvement of calciotropic hormonal regulation was evaluated by studying the response of JEG-3 to 1,25-dihydroxy vitamin D3. Calcium uptake was significantly stimulated in a dose-dependent manner by a 24-h treatment of the cells with 1,25-dihydroxy vitamin D3 (optimal dose approximately 0.5 nM); the CaBP level doubled whereas steady-state CaBP mRNA did not, suggesting that CaBP expression was regulated by 1,25-dihydroxy vitamin D3. These observations strongly suggest that the JEG-3 human choriocarcinoma cells should serve as a convenient in vitro model system for studying the cellular mechanism and regulation of transplacental calcium transport.

219 EFFECT OF THE ENDOCRINE DISRUPTING CHEMICALS ON PLACENTAL TRANSPORT IN BeWo CELLS AS AN IN VITRO MODEL

2015 ◽  
Vol 27 (1) ◽  
pp. 199
Author(s):  
J.-H. Lee ◽  
E.-B. Jeung
Keyword(s):  
In Vitro Model ◽  
Endocrine Disrupting Chemicals ◽  
Reproductive Hormones ◽  
Dependent Manner ◽  
Endocrine Disrupting ◽  
Calcium Cations ◽  
Vitro Model ◽  
Dose Dependent ◽  
Iron And Calcium

The placenta exchanges vital factors, including oxygen, carbon dioxide, copper, iron, calcium cations, and glucose, which are essential to fetal growth. Each molecule is transferred by specific receptors that are located at the cell membrane or in the cytoplasm. Copper, iron, calcium cations, and glucose transfer genes are regulated by estrogens, vitamin D, and human placental lactogen. Regulations of these receptors depend on pregnancy time length and maternal and fetal nutrient environment with various pathways. Some synthetic plastics known as endocrine disrupting chemicals (EDC) have a similar structure to reproductive hormones such as estrogens. Thus, these substances have a potential effect on the expression of genes which are regulated by estrogens or progesterone by interfering their pathways. Having an estrogenic property, EDC interact with oestrogen receptors and elevate or decrease the expression of target genes which are responsible for transporting essential molecules such as copper, iron, and calcium. To examine the effects of EDC exposure during pregnancy, we conducted an in vitro model study using the BeWo human trophoblast cell line. The BeWo cell was treated with well-known EDC, octyl-phenol (OP), nonyl-phenol (NP), and bisphenol A (BPA) in a dose-dependent manner (10–7, 10–6, and 10–5 M) for 24 h. The expression of copper (CTR1, ATP7A), iron (IREG1, HEPH), and calcium transporting genes (PMCA1, TRPV6), were measured by real-time RT–PCR and Western blot. The expression of copper, iron, and calcium transporting genes were elevated in a dose-dependent manner by all well-known EDC, including OP, NP, and BPA, as well as E2. To unveil the mechanism of these elevations of ionic transporting genes, an ERE promoter study will be needed. Taken together, essential cation transporting genes in placenta are modulated by EDC.

Vitamin D-regulated calcium transport in Caco-2 cells: unique in vitro model

1991 ◽  
Vol 260 (2) ◽  
pp. G207-G212 ◽  
Author(s):  
A. R. Giuliano ◽  
R. J. Wood
Keyword(s):  
Vitamin D ◽  
Cell Line ◽  
Calcium Transport ◽  
In Vitro Model ◽  
Colon Adenocarcinoma ◽  
Intestinal Cell ◽  
Maximal Velocity ◽  
Dependent Manner ◽  
Vitro Model

The human colon adenocarcinoma cell line Caco-2 is the only intestinal cell line to differentiate spontaneously in culture exhibiting structural and biochemical characteristics of mature enterocytes and to possess a vitamin D receptor in the fully differentiated state. Transepithelial calcium transport was characterized in differentiated Caco-2 cells grown on permeable filters supports to assess the potential utility of this cell line as an in vitro model to study 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]-induced calcium transport. Calcium transport was increased in a dose-dependent manner by 1,25(OH)2D3. Total calcium transport at different calcium concentrations could be fitted to a modified Michaelis-Menten equation containing a linear transport component. The maximum rate of saturable calcium transport was increased by 4.3-fold (P less than 0.005) in cells treated with 10(-8) M 1,25(OH)2D3. This treatment also increased the apparent buffer calcium concentration that results in half-maximal velocity from 0.4 to 1.3 mM but had no significant effect on nonsaturable calcium transport. Caco-2 cells grown on permeable filter supports provide a unique in vitro human cell culture model to study the mechanism of vitamin D-regulated transepithelial intestinal calcium transport.

scholarly journals Ethanolic extract of ocimum kilimandscharicum linnleaves: phytochemical and in vitro pharmacological activity

2021 ◽  
pp. 106-109
Author(s):  
Yamini N ◽  
Lahari S ◽  
Phani deepthi V
Keyword(s):  
In Vitro Model ◽  
Ethanolic Extract ◽  
Clinical Development ◽  
Dependent Manner ◽  
Thrombolytic Activity ◽  
Standard Drug ◽  
Ethanolic Extracts ◽  
Vitro Model ◽  
Dose Dependent

Using an in vitro model, the anti-thrombolytic efficacy of ethanolic extracts of Ocimum kilimandscharicum Linn was investigated. The researchers discovered that different concentrations of the extract had significant anti-thrombolytic activity in a dose-dependent manner , which was comparable to a standard drug. As a result of the presence of flavonoids and polyphenols in the plant extract, it can be concluded that it has a promising future in the treatment of thrombosis. This knowledge will be useful in the clinical development of thrombolytic therapeutics by identifying more potent anti-thrombolytic principles from natural resources..    

Fibronectin provides a conduit for fibroblast transmigration from collagenous stroma into fibrin clot provisional matrix

1997 ◽  
Vol 110 (7) ◽  
pp. 861-870 ◽  
Author(s):  
D. Greiling ◽  
R.A. Clark
Keyword(s):  
Wound Healing ◽  
In Vitro Model ◽  
Collagen Gel ◽  
Fibrin Clot ◽  
Dependent Manner ◽  
Fibrin Gel ◽  
Concentration Dependent Manner ◽  
Vitro Model ◽  
Early Wound

After injury, the wound space is filled with a fibrin/fibronectin clot containing growth factors released by platelets and monocytes. In response to these factors, fibroblasts migrate into the fibrin clot and contribute to the formation of granulation tissue. The functional mechanisms allowing fibroblasts to leave the collagenous matrix of normal connective tissue and invade the provisional matrix of the fibrin clot have not been fully defined. To investigate these mechanisms we established a new in vitro model which simulates specific aspects of early wound healing, that is, the migration of fibroblasts from a three-dimensional collagen matrix into a fibrin clot. This transmigration could be induced by physiological concentrations of platelet releasate or platelet-derived growth factor BB (PDGF-BB) in a concentration-dependent manner. At 24 hours irradiated fibroblasts invaded the fibrin gel almost as well as non-irradiated cells, indicating that transmigration was independent of proliferation. Plasminogen and its activators appear to be necessary for invasion of the fibrin clot since protease inhibitors decreased the amount of migration. These serine proteases, however, were not necessary for exit from the collagen gel as fibroblasts migrated out of the collagen gel onto a surface coated with fibrin fibrils even in the presence of inhibitors. Removal of fibronectin (FN) from either the collagen gel or the fibrin gel markedly decreased the number of migrating cells, suggesting that FN provides a conduit for transmigration. Cell movement in the in vitro model was inhibited by RGD peptide, and by monoclonal antibodies against the subunits of the alpha5 beta1 and alpha v beta3 integrin receptor. Thus, the functional requirements for fibroblast transmigration from collagen-rich to fibrin-rich matrices, such as occurs in early wound healing, have been partially defined using an in vitro paradigm of this important biologic process.

Interleukin-8 stimulates the migration of human colonic epithelial cells in vitro

1999 ◽  
Vol 97 (3) ◽  
pp. 385-390 ◽  
Author(s):  
Andrew J. WILSON ◽  
Keith BYRON ◽  
Peter R. GIBSON
Keyword(s):  
Cell Migration ◽  
Epithelial Cells ◽  
In Vitro Model ◽  
Interleukin 8 ◽  
Colonic Epithelium ◽  
Dependent Manner ◽  
Colonic Epithelial Cells ◽  
Tnf Α ◽  
Vitro Model

The migration of colonic epithelial cells (restitution) is an important event in the repair of mucosal injuries. Interleukin-8 (IL-8) is a physiological initiator of the chemotactic migration of leucocytes. This study aimed to determine whether IL-8 had a similar effect on migration in an in vitro model of wounded colonic epithelium. Cell migration over 24 h was assessed in circular wounds made in confluent monolayers of the human colon cancer cell line LIM1215. This migration was stimulated in a concentration-dependent manner by IL-8, with maximal effects of approx. 1.75-fold above basal migration. The motogenic effect of IL-8 was mediated independently of effects on cell proliferation. In contrast, it was partially dependent upon gene transcription and protein synthesis and involved the activation of pertussis-toxin-sensitive G-proteins. The short-chain fatty acids, acetate, propionate, butyrate and valerate, the activator of protein kinase C (phorbol-12-myristate-13-acetate) and tumour necrosis factor-α (TNF-α) all stimulated the secretion of IL-8. However, only the motogenic effect of TNF-α was dependent upon IL-8. In conclusion, IL-8 stimulated cell migration in an in vitro model of colonic epithelium, whereas the motogenic effect of at least one physiologically relevant factor was dependent upon an increase in its endogenous levels. If IL-8 stimulates colonic epithelial restitution in vivo, this would have ramifications for the control of repair processes following wounding of the colonic mucosa.

Development and initial application of an in vitro model of apoptosis in rodent cholangiocytes

1997 ◽  
Vol 272 (1) ◽  
pp. G106-G115 ◽  
Author(s):  
F. G. Que ◽  
G. J. Gores ◽  
N. F. LaRusso
Keyword(s):  
In Vitro Model ◽  
Interleukin 1 ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Initial Application ◽  
Histological Data ◽  
Vitro Model ◽  
Induced Apoptosis ◽  
Intracellular Ions

Although histological data suggest that cholangiocytes die by apoptosis in human liver diseases, no information exists on the mechanisms of cholangiocyte apoptosis. Thus our aims were to establish an in vitro model of cholangiocyte apoptosis and to test the hypothesis that changes in intracellular ions would cause apoptosis in cholangiocytes by a protease-sensitive pathway. A large number of proapoptotic agents were ineffective in inducing apoptosis in rat or human cholangiocytes in culture; in contrast, beauvericin, a K+ ionophore, caused apoptosis in both cell lines, despite their expression of Bcl-2. Although beauvericin decreased intracellular K+ and increased intracellular Ca2+, abolishing the K+ gradient did not prevent beauvericin-induced apoptosis; in contrast, omission of extracellular Ca2+ inhibited apoptosis by 42%. The interleukin-1 beta-converting enzyme (ICE) family protease inhibitor, Z-Val-Ala-Asp chloromethylketone, inhibited apoptosis in a concentration-dependent manner. By Northern blot analysis, cholangiocytes expressed the mRNA for three members of the ICE protease family: ICE, ICE/ CED-3 homologue-1 (ICH-1), and cysteine protease P-32 (CPP-32). Cleavage of a substrate for CPP-32-like protease activity, but not a substrate for ICE and ICH-1, increased after beauvericin treatment. In summary, we have established an in vitro model of apoptosis in cholangiocytes. Our data suggest that beauvericin-induced apoptosis occurs by a Ca(2+)-dependent CPP-32 protease-sensitive pathway despite cholangiocyte expression of Bcl-2.

scholarly journals Low Dose BCG Infection as a Model for Macrophage Activation Maintaining Cell Viability

2016 ◽  
Vol 2016 ◽  
pp. 1-17 ◽  
Author(s):  
Leslie Chávez-Galán ◽  
Dominique Vesin ◽  
Denis Martinvalet ◽  
Irene Garcia
Keyword(s):  
Cell Activation ◽  
In Vitro Model ◽  
Low Dose ◽  
Mycobacterial Infection ◽  
Dependent Manner ◽  
Research Fields ◽  
Current Vaccine ◽  
Vitro Model

Mycobacterium bovisBCG, the current vaccine against tuberculosis, is ingested by macrophages promoting the development of effector functions including cell death and microbicidal mechanisms. Despite accumulating reports onM. tuberculosis, mechanisms of BCG/macrophage interaction remain relatively undefined. In vivo, few bacilli are sufficient to establish a mycobacterial infection; however, in vitro studies systematically use high mycobacterium doses. In this study, we analyze macrophage/BCG interactions and microenvironment upon infection with low BCG doses and propose an in vitro model to study cell activation without affecting viability. We show that RAW macrophages infected with BCG at MOI 1 activated higher and sustained levels of proinflammatory cytokines and transcription factors while MOI 0.1 was more efficient for early stimulation of IL-1β, MCP-1, and KC. Both BCG infection doses induced iNOS and NO in a dose-dependent manner and maintained nuclear and mitochondrial structures. Microenvironment generated by MOI 1 induced macrophage proliferation but not MOI 0.1 infection. In conclusion, BCG infection at low dose is an efficient in vitro model to study macrophage/BCG interactions that maintains macrophage viability and mitochondrial structures. This represents a novel model that can be applied to BCG research fields including mycobacterial infections, cancer immunotherapy, and prevention of autoimmunity and allergies.

scholarly journals Morphology, Transcriptomics and In Vitro Model of Skin from Polar Cod (Boreogadus Saida) and Atlantic Cod (Gadus Morhua)

Fishes ◽  
2020 ◽  
Vol 5 (4) ◽  
pp. 34
Author(s):  
Elisabeth Ytteborg ◽  
Øyvind Johannes Hansen ◽  
Vibeke Høst ◽  
Sergey Afanasyev ◽  
Ireen Vieweg ◽  
...  
Keyword(s):  
Gadus Morhua ◽  
In Vitro Model ◽  
Atlantic Cod ◽  
Polar Cod ◽  
Dependent Manner ◽  
Boreogadus Saida ◽  
Polar Cod Boreogadus Saida ◽  
Vitro Model ◽  
Skin Healing

Fish skin is a multifunctional barrier tissue with high regeneration capacity that interacts with the surrounding environment and provides protection. Functional importance, high complexity and activity make skin an attractive tissue for studying the effects of environmental challenges and chemical stressors in fish. The aim of this work was to characterize skin from polar cod (Boreogadus saida) and Atlantic cod (Gadus morhua), and to test cod skin as an in vitro model in exposure studies. Both species have similar skin structures including epidermis, mucous cells, club cells and scales. However, microarchitectural differences were detected; Atlantic cod has a smooth epidermal surface and overlapping scales, whereas polar cod has a folded outer surface and discontinuous scales. Genome-wide microarray found 6.5k genes with expression differences, which suggested more active turnover of proteins, proliferation and motility of cells in skin of polar cod. Hydrogen peroxide (H2O2) was used to examine skin responses. Transcriptome response was stronger in the skin of polar cod, with 155 differentially expressed genes. The skin from Atlantic cod was further used to develop a cell culture. H2O2 decreased the cell migration rate in a dose-dependent manner, which could indicate reduced skin healing capacity. The results revealed novel skin structures and confirmed that the skin from cod is a promising tissue for evaluation of stressors.

scholarly journals Hyperosmolar Potassium (K+) Treatment Suppresses Osteoarthritic Chondrocyte Catabolic and Inflammatory Protein Production in a 3-Dimensional In Vitro Model

Cartilage ◽  
2017 ◽  
Vol 10 (2) ◽  
pp. 186-195 ◽  
Author(s):  
Josh Erndt-Marino ◽  
Erik Trinkle ◽  
Mariah S. Hahn
Keyword(s):  
Inflammatory Mediators ◽  
Protein Production ◽  
In Vitro Model ◽  
Culture Media ◽  
Dependent Manner ◽  
Proof Of Concept ◽  
3 Dimensional ◽  
Inflammatory Protein ◽  
Vitro Model

Objective The main goal of this study was to provide a proof-of-concept demonstrating that hyperosmolar K+ solutions can limit production of catabolic and inflammatory mediators in human osteoarthritic chondrocytes (OACs). Methods A 3-dimensional in vitro model with poly(ethylene glycol) diacrylate (PEGDA) hydrogels was used. Catabolic and pro-inflammatory protein production from encapsulated OACs was assessed following culture for 1 or 7 days in the presence or absence of 80 mM K+ gluconate, 80 mM sodium (Na+) gluconate, or 160 mM sucrose, each added to culture media (final osmolarity ~490 mOsm). Results Relative to untreated controls, OACs treated with hyperosmolar (80 mM Na+ gluconate or 160 mM sucrose) solutions produced lower levels of catabolic and inflammatory mediators in a marker- and time-dependent manner (i.e., MMP-9 after 1 day; MCP-1 after 7 days ( P ≤ 0.015)). In contrast, OAC treatment with 80 mM K+ gluconate reduced catabolic and inflammatory mediators to a greater extent (both the number of markers and degree of suppression) relative to untreated, Na+ gluconate, or sucrose controls (i.e., MMP-3, -9, -13, TIMP-1, MCP-1, and IL-8 after 1 day; MMP-1, -3, -9, -13, TIMP-1, MCP-1, and IL-8 after 7 days ( P ≤ 0.029). Conclusions Hyperosmolar K+ solutions are capable of attenuating protein production of catabolic and inflammatory OA markers, providing the proof-of-concept needed for further development of a K+-based intra-articular injection for OA treatment. Moreover, K+ performed significantly better than Na+- or sucrose-based solutions, supporting the application of K+ toward improving irrigation solutions for joint surgery.

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