Salivary epithelial cells in primary culture: characterization of their growth and functional properties

1976 ◽  
Vol 20 (1) ◽  
pp. 149-165 ◽  
Author(s):  
C.B. Wigley ◽  
L.M. Franks
Keyword(s):  
Epithelial Cells ◽  
Primary Culture ◽  
Initial Period ◽  
Explant Culture ◽  
Submandibular Salivary Gland ◽  
Salivary Gland Cells ◽  
Primary Explant

Mouse submandibular salivary gland cells were grown in primary explant culture. After an initial period of degeneration within the explant, surviving epithelial cells proliferated rapidly and duct-like structures recolonized the explant. Autoradiographic studies showed that a peak of DNA synthesis occurred after 4 days in vitro and that proliferation was enhanced by insulin and hydrocortisone. These cells retained specialized secretory function (protease activity) for at least 2 weeks in vitro. This enzyme is a differentiated product of granular tubule cells in vivo. Between 6 and 10 days, explants attached to the substrate. An outgrowth developed, consisting largely of ultrastructurally identifiable epithelial cells which formed pseudoglandular structures in the monolayer. Epithelium survived for over 6 months in primary culture but could not be serially transferred. Secondary cultures were rapidly overgrown by mesenchymal cells.

scholarly journals Functional and Genomic Characterization of Ligilactobacillus salivarius TUCO-L2 Isolated From Lama glama Milk: A Promising Immunobiotic Strain to Combat Infections

2020 ◽  
Vol 11 ◽  
Author(s):  
Sandra Quilodrán-Vega ◽  
Leonardo Albarracin ◽  
Flavia Mansilla ◽  
Lorena Arce ◽  
Binghui Zhou ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Functional Characterization ◽  
Comparative Genomic ◽  
Cytokines And Chemokines ◽  
Isolation And Characterization ◽  
Intestinal Pathogens ◽  
Lama Glama

Potential probiotic or immunobiotic effects of lactic acid bacteria (LAB) isolated from the milk of the South American camelid llama (Lama glama) have not been reported in published studies. The aim of the present work was to isolate beneficial LAB from llama milk that can be used as potential probiotics active against bacterial pathogens. LAB strains were isolated from llama milk samples. In vitro functional characterization of the strains was performed by evaluating the resistance against gastrointestinal conditions and inhibition of the pathogen growth. Additionally, the adhesive and immunomodulatory properties of the strains were assessed. The functional studies were complemented with a comparative genomic evaluation and in vivo studies in mice. Ligilactobacillus salivarius TUCO-L2 showed enhanced probiotic/immunobiotic potential compared to that of other tested strains. The TUCO-L2 strain was resistant to pH and high bile salt concentrations and demonstrated antimicrobial activity against Gram-negative intestinal pathogens and adhesion to mucins and epithelial cells. L. salivarius TUCO-L2 modulated the innate immune response triggered by Toll-like receptor (TLR)-4 activation in intestinal epithelial cells. This effect involved differential regulation of the expression of inflammatory cytokines and chemokines mediated by the modulation of the negative regulators of the TLR signaling pathway. Moreover, the TUCO-L2 strain enhanced the resistance of mice to Salmonella infection. This is the first report on the isolation and characterization of a potential probiotic/immunobiotic strain from llama milk. The in vitro, in vivo, and in silico investigation performed in this study reveals several research directions that are needed to characterize the TUCO-L2 strain in detail to position this strain as a probiotic or immunobiotic that can be used against infections in humans or animals, including llama.

scholarly journals Characterization of Candidate Live Oral Salmonella typhi Vaccine Strains Harboring Defined Mutations inaroA, aroC, and htrA

1999 ◽  
Vol 67 (2) ◽  
pp. 700-707 ◽  
Author(s):  
David C. Lowe ◽  
Tor C. Savidge ◽  
Derek Pickard ◽  
Lars Eckmann ◽  
Martin F. Kagnoff ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Salmonella Typhi ◽  
Scid Mice ◽  
Intestinal Epithelial ◽  
Human Intestinal Epithelial Cells ◽  
U937 Macrophage ◽  
Ht 29

ABSTRACT The properties of two candidate Salmonella typhi-based live oral typhoid vaccine strains, BRD691 (S. typhi Ty2 harboring mutations in aroA and aroC) and BRD1116 (S. typhi Ty2 harboring mutations inaroA, aroC, and htrA), were compared in a number of in vitro and in vivo assays. BRD1116 exhibited an increased susceptibility to oxidative stress compared with BRD691, but both strains were equally resistant to heat shock. Both strains showed a similar ability to invade Caco-2 and HT-29 epithelial cells and U937 macrophage-like cells, but BRD1116 was less efficient at surviving in epithelial cells than BRD691. BRD1116 and BRD691 were equally susceptible to intracellular killing within U937 cells. Similar findings were demonstrated in vivo, with BRD1116 being less able to survive and translocate to secondary sites of infection when inoculated into the lumen of human intestinal xenografts in SCID mice. However, translocation of BRD1116 to spleens and livers in SCID mice occurred as efficiently as that of BRD691 when inoculated intraperitonally. The ability of BRD1116 to increase the secretion of interleukin-8 following infection of HT-29 epithelial cells was comparable to that of BRD691. Therefore, loss of the HtrA protease inS. typhi does not seem to alter its ability to invade epithelial cells or macrophages or to induce proinflammatory cytokines such as IL-8 but significantly reduces intracellular survival in human intestinal epithelial cells in vitro and in vivo.

Vimentin affects localization and activity of sodium-glucose cotransporter SGLT1 in membrane rafts

2002 ◽  
Vol 115 (4) ◽  
pp. 713-724
Author(s):  
Isabelle Runembert ◽  
Guillaume Queffeulou ◽  
Pierre Federici ◽  
François Vrtovsnik ◽  
Emma Colucci-Guyon ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Primary Culture ◽  
Glucose Transport ◽  
Cholesterol Metabolism ◽  
Cell Structure ◽  
Cholesterol Content ◽  
Membrane Microdomains ◽  
Transport Activity

It has been reported that vimentin, a cytoskeleton filament that is expressed only in mesenchymal cells after birth, is re-expressed in epithelial cells in vivo under pathological conditions and in vitro in primary culture. Whether vimentin re-expression is only a marker of cellular dedifferentiation or is instrumental in the maintenance of cell structure and/or function is a matter of debate. To address this issue, we used renal proximal tubular cells in primary culture from vimentin-null mice (Vim-/-) and from wild-type littermates (Vim+/+). The absence of vimentin did not affect cell morphology, proliferation and activity of hydrolases, but dramatically decreased Na-glucose cotransport activity. This phenotype was associated with a specific reduction of SGLT1 protein in the detergent-resistant membrane microdomains (DRM). In Vim+/+cells, disruption of these microdomains by methyl-β-cyclodextrin decreased SGLT1 protein abundance in DRM, a change that was paralleled by a decrease of Na-glucose transport activity. Importantly, we showed that vimentin is located to DRM, but it disappeared after methyl-β-cyclodextrin treatment. In Vim-/- cells,supplementation of cholesterol with cholesterol-methyl-β-cyclodextrin complexes completely restored Na-glucose transport activity. Interestingly,neither cholesterol content nor cholesterol metabolism changed in Vim-/- cells. Our results are consistent with the view that re-expression of vimentin in epithelial cells could be instrumental to maintain the physical state of rafts and, thus, the function of DRM-associated proteins.

In Vitro and In Vivo Characterization of Pigment Epithelial Cells Differentiated from Primate Embryonic Stem Cells

2004 ◽  
Vol 45 (3) ◽  
pp. 1020 ◽  
Author(s):  
Masatoshi Haruta ◽  
Yoshiki Sasai ◽  
Hiroshi Kawasaki ◽  
Kaori Amemiya ◽  
Sotaro Ooto ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Epithelial Cells ◽  
Embryonic Stem ◽  
Pigment Epithelial ◽  
In Vivo Characterization ◽  
Pigment Epithelial Cells

Rapid, Refined, and Robust Method for Expression, Purification, and Characterization of Recombinant Human Amyloid-beta M1-42

2019 ◽  
Author(s):  
Priya Prakash ◽  
Travis Lantz ◽  
Krupal P. Jethava ◽  
Gaurav Chopra
Keyword(s):  
Amyloid Beta ◽  
Western Blots ◽  
Force Microscopy ◽  
E Coli ◽  
Atomic Force ◽  
Set Up ◽  
Short Time

Amyloid plaques found in the brains of Alzheimer’s disease (AD) patients primarily consists of amyloid beta 1-42 (Ab42). Commercially, Ab42 is synthetized using peptide synthesizers. We describe a robust methodology for expression of recombinant human Ab(M1-42) in Rosetta(DE3)pLysS and BL21(DE3)pLysS competent E. coli with refined and rapid analytical purification techniques. The peptide is isolated and purified from the transformed cells using an optimized set-up for reverse-phase HPLC protocol, using commonly available C18 columns, yielding high amounts of peptide (~15-20 mg per 1 L culture) in a short time. The recombinant Ab(M1-42) forms characteristic aggregates similar to synthetic Ab42 aggregates as verified by western blots and atomic force microscopy to warrant future biological use. Our rapid, refined, and robust technique to purify human Ab(M1-42) can be used to synthesize chemical probes for several downstream in vitro and in vivo assays to facilitate AD research.

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