scholarly journals Higher percentage of horse serum in culture media blocks attachment of PC12 cells

BioTechniques ◽  
2019 ◽  
Vol 67 (6) ◽  
pp. 256-258
Author(s):  
Jennifer L Meth ◽  
Alan R Schoenfeld
Keyword(s):  
1985 ◽  
Vol 101 (4) ◽  
pp. 1487-1491 ◽  
Author(s):  
H Katow ◽  
M Hayashi

We studied the effect of fibronectin (FN) on the behavior of primary mesenchyme cells isolated from sea urchin mesenchyme blastulae in vitro using a time-lapse technique. The migration of isolated primary mesenchyme cells reconstituted in seawater and horse serum is dependent on the presence or absence of exogenous FN in the culture media. The cells in FN, 4 and 40 micrograms/ml, show a high percentage of migration and migrate long distances, whereas a higher concentration of FN at 400 micrograms/ml tends to inhibit migration.


2000 ◽  
Vol 38 (5) ◽  
pp. 1984-1987 ◽  
Author(s):  
Xiuping Jiang ◽  
Michael P. Doyle

The growth response of Helicobacter pylori in broth was determined in the presence of ferrous sulfate, sodium pyruvate, and mucin (porcine stomach). The addition of either ferrous sulfate and sodium pyruvate or mucin to brain heart infusion broth with 7% horse serum (BHI-HS) enhanced the growth of H. pylori. The best growth of strain NB2-1, which was the slowest growing of 10 H. pylori strains evaluated, occurred in the presence of 0.05% ferrous sulfate and 0.05% sodium pyruvate. The addition of 0.3% mucin to BHI-HS reduced the lag time of H. pylori by 48 h and enhanced the growth. On the basis of the results for 10 H. pylori strains, the combination of ferrous sulfate (0.025%), sodium pyruvate (0.025%), and mucin (0.15%) in BHI-HS counteracted the inhibitory effects of the antibiotics used in culture media for selective growth of H. pylori. Results suggest that these supplements may be useful for enhancement of the growth of H. pylori in enrichment media.


2017 ◽  
Vol 34 (3) ◽  
pp. 139-145 ◽  
Author(s):  
Xuemei Fang ◽  
Chao Wu ◽  
Hongxia Li ◽  
Weifeng Yuan ◽  
Xin Wang

Perfluorononanoic acid (PFNA) is one of the major perfluorinated compounds found in both biological and abiotic samples and has recently been demonstrated to cause neurobehavioral defects in mammals. In this study, pheochromocytoma-12 (PC12) cells were exposed to various doses of PFNA to explore the cytotoxicity of PFNA to neurons and the possible mechanisms underlying these effects. The results showed that exposure to PFNA dose-dependently decreased the viability of PC12 cells and increased the release of lactate dehydrogenase into cell culture media. Exposure to PFNA increased the malondialdehyde content and decreased the total antioxidant capacity and glutathione peroxidase activity in PC12 cell culture supernatants. Exposure to PFNA increased the intracellular calcium level and upregulated the Ca2+/calmodulin-dependent protein kinase II (CaMKII) expression in PC12 cells. PFNA also decreased Bcl-2 expression and increased Bax expression in PC12 cells. These results suggested that exposure to PFNA elevated the intracellular calcium level and activated the CaMKII signaling pathway, which may aggravate oxidative stress in PC12 cells and lead to cell damage or cell apoptosis.


Author(s):  
Shinji Miyazaki ◽  
Yuji Omiya ◽  
Kazushige Mizoguchi

Abstract Dementia is exacerbated by loss of appetite and amotivation, recent studies have indicated that ninjin'yoeito improves anorexia and amotivation. Previous studies suggest that ninjin'yoeito inhibits dopamine-metabolizing enzymes and enhances dopamine signaling. However, whether ninjin'yoeito increases dopamine content in living cells remains unclear. Here, PC12 cells were used to examine whether ninjin'yoeito affects the dopamine metabolic pathway. Dopamine content significantly increased 3 h after treatment ninjin'yoeito extract. Concomitantly, the levels of 3-methoxytyramine and 3,4-dihydroxyphenylacetic acid was significantly reduced. The effects of components of ninjin'yoeito on the dopamine metabolic pathway were also assessed. Treatment with onjisaponin B, nobiletin, and schisandrin, the ingredients of Polygalae Radix, Citri Unshiu Pericarpium, and Schisandrae Fructus increased dopamine content and decreased its metabolite content in the culture media. Our findings suggest that ninjin'yoeito improves anorexia and amotivation by inhibiting metabolic enzyme and increasing the dopamine content in cells.


1976 ◽  
Vol 4 (3) ◽  
pp. 208-213
Author(s):  
D B Louria ◽  
T Kaminski ◽  
R Kapila ◽  
F Tecson ◽  
L Smith

Specimens from 300 patients were studied using five to nine aerobic and anaerobic culture media, including five that were hypertonic, Groups studied included fever of unknown origin, suspected endocarditis, endocarditis during therapy, bacteremia during therapy, abscess and cellulitis, presumed infectious arthritis, renal transplantation during rejection, collagen disease, sarcoidosis, lymphoma, and colitis. Isolates in hypertonic media were reverted to parent form by agar passage. In only 5% of these selected cases were organisms found in hypertonic, but not conventional, media that appeared on the basis of repeated isolation and/or serological studies to come from the patient. Nine of the 16 appeared to be of major significance. The two groups in which use of highly enriched, hypertonic media seemed most helpful were suspected endocarditis and undefined meningitis with negative cultures using standard media. The most effective of the hypertonic media used was 0.3 M sucrose in brain heart infusion with 20% horse serum. In most instances, the organism grew only in the hypertonic sucrose, and in most cases it appeared in conventional rather than aberrant form. Hypertonic media, especially 0.3 M sucrose, are of substantial helpin a small number of carefully selected cases.


Author(s):  
Marek Malecki ◽  
James Pawley ◽  
Hans Ris

The ultrastructure of cells suspended in physiological fluids or cell culture media can only be studied if the living processes are stopped while the cells remain in suspension. Attachment of living cells to carrier surfaces to facilitate further processing for electron microscopy produces a rapid reorganization of cell structure eradicating most traces of the structures present when the cells were in suspension. The structure of cells in suspension can be immobilized by either chemical fixation or, much faster, by rapid freezing (cryo-immobilization). The fixation speed is particularly important in studies of cell surface reorganization over time. High pressure freezing provides conditions where specimens up to 500μm thick can be frozen in milliseconds without ice crystal damage. This volume is sufficient for cells to remain in suspension until frozen. However, special procedures are needed to assure that the unattached cells are not lost during subsequent processing for LVSEM or HVEM using freeze-substitution or freeze drying. We recently developed such a procedure.


Author(s):  
E. S. Boatman ◽  
G. E. Kenny

Information concerning the morphology and replication of organism of the family Mycoplasmataceae remains, despite over 70 years of study, highly controversial. Due to their small size observations by light microscopy have not been rewarding. Furthermore, not only are these organisms extremely pleomorphic but their morphology also changes according to growth phase. This study deals with the morphological aspects of M. pneumoniae strain 3546 in relation to growth, interaction with HeLa cells and possible mechanisms of replication.The organisms were grown aerobically at 37°C in a soy peptone yeast dialysate medium supplemented with 12% gamma-globulin free horse serum. The medium was buffered at pH 7.3 with TES [N-tris (hyroxymethyl) methyl-2-aminoethane sulfonic acid] at 10mM concentration. The inoculum, an actively growing culture, was filtered through a 0.5 μm polycarbonate “nuclepore” filter to prevent transfer of all but the smallest aggregates. Growth was assessed at specific periods by colony counts and 800 ml samples of organisms were fixed in situ with 2.5% glutaraldehyde for 3 hrs. at 4°C. Washed cells for sectioning were post-fixed in 0.8% OSO4 in veronal-acetate buffer pH 6.1 for 1 hr. at 21°C. HeLa cells were infected with a filtered inoculum of M. pneumoniae and incubated for 9 days in Leighton tubes with coverslips. The cells were then removed and processed for electron microscopy.


Author(s):  
Douglas R. Keene ◽  
Gregory P. Lunstrum ◽  
Patricia Rousselle ◽  
Robert E. Burgeson

A mouse monoclonal antibody produced from collagenase digests of human amnion was used by LM and TEM to study the distribution and ultrastructural features of an antigen present in epithelial tissues and in cultured human keratinocytes, and by immunoaffinity chromatography to partially purify the antigen from keratinocyte cell culture media.By immunofluorescence microscopy, the antigen displays a tissue distribution similar to type VII collagen; positive staining of the epithelial basement membrane is seen in skin, oral mucosa, trachea, esophagus, cornea, amnion and lung. Images from rotary shadowed preparations isolated by affinity chromatography demonstrate a population of rod-like molecules 107 nm in length, having pronounced globular domains at each end. Polyacrylamide gel electrophoresis suggests that the size of this molecule is approximately 440kDa, and that it is composed of three nonidentical chains disulfide bonded together.


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