Primary Culture of Strial Marginal Cells of Guinea Pig Cochlea: Growth, Morphologic Features, and Characterization

1991 ◽  
Vol 100 (12) ◽  
pp. 999-1006 ◽  
Author(s):  
Josué Achouche ◽  
An H. Wu ◽  
Der S. Liu ◽  
Patrice Tran Ba Huy
Keyword(s):  
Guinea Pig ◽  
Primary Culture ◽  
Cultured Cells ◽  
Stria Vascularis ◽  
Primary Cultures ◽  
Suitable Model ◽  
Type I ◽  
Cellular Mechanisms ◽  
Electron Microscopic ◽  
Marginal Cells

To further investigate the cellular mechanisms involved in the formation of endolymph, primary cultures of marginal cells of guinea pig were established. Minute explants obtained by mechanical dissociation of stria vascularis were plated on collagen type I precoated impermeable substrate in serum-free, hormone-supplemented medium. A confluent layer of epithelial-like cells was obtained within 2 weeks. The cultured cells formed domes, demonstrating that they retain some of their transepithelial properties. Polarization was also suggested by electron microscopic observation of apical microvilli and tight junctions. Immunohistochemical methods revealed that the cultured cells coexpressed cytokeratin and vimentin, demonstrating their epithelial origin, although some degree of dedifferentiation occurred. Thus, a primary culture of marginal cells can be established that may be a suitable model for an in-depth investigation of the function of the marginal cells.

Movement of Monovalent Ions across the Membranes of Marginal Cells of the Stria vascularis in the Guinea Pig Cochlea

ORL ◽  
1993 ◽  
Vol 55 (2) ◽  
pp. 61-67 ◽  
Author(s):  
Shizuo Komune ◽  
Takashi Nakagawa ◽  
Kazutaka Hisashi ◽  
Takashi Kimituki ◽  
Takuya Uemura
Keyword(s):  
Guinea Pig ◽  
Stria Vascularis ◽  
Guinea Pig Cochlea ◽  
Monovalent Ions ◽  
Marginal Cells

scholarly journals Macromolecular Organization and In Vitro Growth Characteristics of Scaffold-free Neocartilage Grafts

2007 ◽  
Vol 55 (8) ◽  
pp. 853-866 ◽  
Author(s):  
Anthony J. Hayes ◽  
Amanda Hall ◽  
Liesbeth Brown ◽  
Ross Tubo ◽  
Bruce Caterson
Keyword(s):  
Articular Cartilage ◽  
Type I Collagen ◽  
Primary Cultures ◽  
Surface Zone ◽  
Type I ◽  
Focal Lesions ◽  
Electron Microscopic ◽  
Chondrocyte Phenotype ◽  
Macromolecular Organization

Recent advances in tissue engineering offer considerable promise for the repair of focal lesions in articular cartilage. Here we describe (1) the macromolecular organization of tissue-engineered neocartilage grafts at light and electron microscopic levels, (2) their in vitro development, and (3) the effect of chondrocyte dedifferentiation, induced by monolayer expansion, on their resultant structure. We show that grafts produced from primary cultures of chondrocytes are hyaline in appearance with identifiable zonal strata as evidenced by cell morphology, matrix organization, and immunohistochemical composition. Like native articular cartilage, their surface zone contains type I collagen, surface zone proteoglycan, biglycan and decorin with type II collagen, aggrecan, chondroitin sulfate, chondroitin-4-sulfate, and keratan sulfate, becoming more prominent with depth. Assessment of cell viability by Live/Dead staining and cell-cycle analysis with BrDU suggest that the in vitro tissue has a high cellular turnover and develops through both appositional and interstitial growth mechanisms. Meanwhile, cell-tracker studies with CMFDA (5-chloromethyl-fluorescein diacetate) demonstrate that cell sorting in vitro is not involved in their zonal organization. Finally, passage expansion of chondrocytes in monolayer culture causes progressive reductions in graft thickness, loss of zonal architecture, and a more fibrocartilaginous tissue histology, consistent with a dedifferentiating chondrocyte phenotype. (J Histochem Cytochem 55: 853–866, 2007)

Voltage-activated K channel in luminal membrane of marginal cells of stria vascularis dissected from guinea pig

1994 ◽  
Vol 80 (1) ◽  
pp. 86-92 ◽  
Author(s):  
Hiroshi Sunose ◽  
Katsuhisa Ikeda ◽  
Masaaki Suzuki ◽  
Tomonori Takasaka
Keyword(s):  
Guinea Pig ◽  
Stria Vascularis ◽  
K Channel ◽  
Luminal Membrane ◽  
Marginal Cells

scholarly journals Regulation of collagen production and collagen mRNA amounts in fibroblasts in response to culture conditions

1986 ◽  
Vol 239 (1) ◽  
pp. 179-183 ◽  
Author(s):  
S R Quinones ◽  
D S Neblock ◽  
R A Berg
Keyword(s):  
Type I Collagen ◽  
Cultured Cells ◽  
Primary Cultures ◽  
Culture Conditions ◽  
Type I ◽  
Collagen Production ◽  
Tendon Cells ◽  
Chain Synthesis ◽  
Polypeptide Chains

Collagen synthesis and mRNA amounts for the alpha 1 and alpha 2 polypeptide chains of Type I collagen were measured in embryonic-chick tendons and in tendon cells both in suspension and in primary cultures. The percentage of protein production represented by collagen in suspension-cultured cells was initially the same as in the intact tendon; however, on an hourly basis, there was actually a steady decline in collagen production by suspended cells. Collagen production in primary cultures of chick tendon fibroblasts was decreased when compared with intact tendon, even though ascorbate-supplemented primary cultures were able to maintain higher rates of collagen production than were non-supplemented cultures. The amounts of mRNA for alpha 1(I) and alpha 2(I) polypeptide chains of collagen responded in similar fashions to different culture conditions and were compared with the amounts of mRNA for beta-actin. In primary cultures the available alpha 1 and alpha 2 collagen mRNAs support proportionately higher collagen production than in the intact tendon. However, the ratio of alpha 1/alpha 2 mRNA and polypeptide-chain synthesis did not remain 2:1, but increased with the concomitant production of Type I trimers composed of three alpha 1 chains. Removal of fibroblasts from their environment in vivo appears to alter the amounts of mRNA for alpha 1 and alpha 2 chains and to alter the utilization of those mRNAs for polypeptide synthesis.

Electrical, contractile, and ultrastructural properties of adult rat and guinea-pig ventricular myocytes in long-term primary cultures

1989 ◽  
Vol 67 (7) ◽  
pp. 740-750 ◽  
Author(s):  
M. Horackova ◽  
C. Mapplebeck
Keyword(s):  
Guinea Pig ◽  
Cultured Cells ◽  
Ventricular Myocytes ◽  
Contractile Activity ◽  
Primary Cultures ◽  
Morphological Characteristics ◽  
Ultrastructural Organization ◽  
Adult Rat ◽  
Intercellular Contacts

The electrical, contractile, and morphological characteristics of ventricular myocytes isolated from adult rat and guinea-pig hearts and maintained in cultures for 7–24 days are described. These cultured cells form different networks, depending on the species, when plated at certain density and maintained under specific conditions; the cells within the networks appear to be electrically coupled. Their resting and action potentials, their contractile activity (shortenings), and their pharmacological responses qualitatively resemble those of freshly isolated myocytes. Cultured cells from both species exhibit near-normal ultrastructural organization of sarcomeres, myofilaments, and mitochondria, as well as formation of intercellular contacts, or gap junctions. These data indicate that cultured adult rat and guinea-pig myocardial cells that make intercellular contacts possess electrical, contractile, and ultrastructural properties and responses to pharmacological agents similar to those of the respective adult myocardial tissues and the functionally intact freshly isolated cells from which these cultures are prepared. Thus, this study indicates that long-term cultures (7–24 days) of networked cardiac myocytes could be used as a valuable experimental model in various investigations of excitation–contraction coupling in cardiac muscle.Key words: cultured adult cardiomyocytes, contractility, electrical activity, ultrastructure, long-term primary cultures.

Helicobacter pylori lipopolysaccharide activates Rac1 and transcription of NADPH oxidase Nox1 and its organizer NOXO1 in guinea pig gastric mucosal cells

2005 ◽  
Vol 288 (2) ◽  
pp. C450-C457 ◽  
Author(s):  
Tsukasa Kawahara ◽  
Motoyuki Kohjima ◽  
Yuki Kuwano ◽  
Hisano Mino ◽  
Shigetada Teshima-Kondo ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Guinea Pig ◽  
Nadph Oxidase ◽  
Primary Cultures ◽  
Transcriptional Level ◽  
Type I ◽  
Gastric Mucosal Cells ◽  
Mucosal Cells ◽  
H Pylori ◽  
Constitutively Active

Primary cultures of guinea pig gastric mucosal cells express NADPH oxidase 1 (Nox1), a homolog of gp91 phox, and produce superoxide anion (O2−) at a rate of ∼100 nmol·mg protein−1·h−1 in response to Helicobacter pylori ( H. pylori) lipopolysaccharide (LPS) from virulent type I strains. The upregulated O2− production also enhances H. pylori LPS-stimulated tumor necrosis factor-α or cyclooxygenase-2 mRNA expression, which suggests a potential role for Nox1 in the pathogenesis of H. pylori-associated diseases. The H. pylori LPS-stimulated O2− production in cultured gastric mucosal cells was inhibited by actinomycin D as well as cycloheximide, suggesting that the induction is regulated at the transcriptional level. The LPS treatment not only increased the Nox1 mRNA to a greater extent but also induced expression of the message-encoding, Nox-organizing protein 1 (NOXO1), a novel p47 phox homolog required for Nox1 activity. In addition, H. pylori LPS activated Rac1; i.e., it converted Rac1 to the GTP-bound state. A phosphoinositide 3-kinase inhibitor, LY-294002, blocked H. pylori LPS-induced Rac1 activation and O2− generation without interfering with the expression of Nox1 and NOXO1 mRNA. O2− production inhibited by LY-294002 was completely restored by transfection of an adenoviral vector encoding a constitutively active Rac1 but not an inactive Rac1 or a constitutively active Cdc42. These findings indicate that Rac1 plays a crucial role in Nox1 activation. Thus the H. pylori LPS-stimulated O2− production in gastric mucosal cells appears to require two distinct events: 1) transcriptional upregulation of Nox1 and NOXO1 and 2) activation of Rac1.

Pepsinogen secretion from dispersed chief cells from guinea pig stomach

1984 ◽  
Vol 247 (1) ◽  
pp. G95-G104 ◽  
Author(s):  
J. P. Raufman ◽  
V. E. Sutliff ◽  
D. K. Kasbekar ◽  
R. T. Jensen ◽  
J. D. Gardner
Keyword(s):  
Guinea Pig ◽  
Incubation Temperature ◽  
Suitable Model ◽  
Cellular Mechanisms ◽  
Gastric Glands ◽  
Chief Cells ◽  
Pepsinogen Secretion ◽  
Mucosal Cells ◽  
Percoll Density Gradient ◽  
Guinea Pig Stomach

In the present study we examined the actions of various secretagogues on pepsinogen secretion from freshly dispersed chief cells prepared from guinea pig stomach. Chief cells were obtained by preparing dispersed gastric glands, subjecting the glands to mechanical disruption in the presence of EGTA, and fractionating the resulting mucosal cells on a Percoll density gradient. Chief cells constituted 90% of the final cell suspension and cell viability was 99%. In these cells, pepsinogen secretion was stimulated by agents whose actions are probably mediated by calcium: carbachol, cholecystokinin, and A23187. Pepsinogen secretion was also stimulated by agents whose actions are probably mediated by cAMP: secretin, vasoactive intestinal peptide, and 8-bromo-cAMP. Reducing the incubation temperature from 37 degrees to 4 degrees C or adding carbonyl cyanide m-chlorophenylhydrazone abolished secretagogue-induced pepsinogen secretion. These results indicate that freshly dispersed chief cells from guinea pig stomach are responsive to secretagogues and provide a suitable model for investigating cellular mechanisms of secretagogue-induced pepsinogen secretion.

Distribution and significance of endothelin 1 in guinea pig cochlear lateral wall

2007 ◽  
Vol 121 (8) ◽  
pp. 721-724 ◽  
Author(s):  
D-Y Xu ◽  
Y-D Tang ◽  
S-X Liu ◽  
J Liu
Keyword(s):  
Guinea Pig ◽  
Stria Vascularis ◽  
Lateral Wall ◽  
Complex Method ◽  
Spiral Ligament ◽  
Basal Cells ◽  
Endothelin 1 ◽  
Marginal Cells ◽  
Spiral Ligament Fibrocytes ◽  
Avidin Biotin Complex

AbstractEndothelin 1 is a vasoconstrictive peptide with many biological functions. To investigate the distribution of endothelin 1 in guinea pig cochlear lateral wall and the significance of endothelin 1 in maintaining cochlear homeostasis, the immunohistochemistry avidin biotin complex method was applied by using rabbit anti-endothelin 1 polyclonal antibody as primary antibody. Endothelin-1-like activities were detected in the marginal cells, spiral prominence epithelial cells, outer sulcus cells, stria vascularis capillaries, basal cells and spiral ligament fibrocytes.These results suggest that endothelin 1 may play an important role in maintaining cochlear homeostasis.

Quinacrine staining of marginal cells in the stria vascularis of the guinea-pig cochlea: a possible source of extracellular ATP?

1995 ◽  
Vol 90 (1-2) ◽  
pp. 97-105 ◽  
Author(s):  
P.N. White ◽  
P.R. Thorne ◽  
G.D. Housley ◽  
B. Mockett ◽  
T.E. Billett ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Stria Vascularis ◽  
Extracellular Atp ◽  
Guinea Pig Cochlea ◽  
Marginal Cells
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