Generation of primary cells from ADPKD and normal human kidneys

2019 ◽  
pp. 1-23 ◽  
Author(s):  
Darren P. Wallace ◽  
Gail A. Reif
Keyword(s):  
Primary Cells ◽  
Normal Human

scholarly journals The expression of the eotaxins IL-6 and CXCL8 in human epithelial cells from various levels of the respiratory tract

2013 ◽  
Vol 18 (4) ◽  
Author(s):  
Magdalena Paplińska-Goryca ◽  
Patrycja Nejman-Gryz ◽  
Ryszarda Chazan ◽  
Hanna Grubek-Jaworska
Keyword(s):  
Epithelial Cells ◽  
Respiratory Tract ◽  
Airway Epithelial Cells ◽  
Small Airway ◽  
Primary Cells ◽  
Human Epithelial Cells ◽  
Tnf Α ◽  
Normal Human ◽  
Ifn Γ ◽  
Pre Treatment

AbstractAirway epithelium acts as multifunctional site of response in the respiratory tract. Epithelial activity plays an important part in the pathophysiology of obstructive lung disease. In this study, we compare normal human epithelial cells from various levels of the respiratory tract in terms of their reactivity to pro-allergic and pro-inflammatory stimulation. Normal human nasal, bronchial and small airway epithelial cells were stimulated with IL-4 and IL-13. The expressions of the eotaxins IL-6 and CXCL8 were evaluated at the mRNA and protein levels. The effects of pre-treatment with IFN-γ on the cell reactivity were measured, and the responses to TNF-α, LPS and IFN-γ were evaluated. All of the studied primary cells expressed CCL26, IL-6 and IL-8 after IL-4 or IL-13 stimulation. IFN-γ pre-treatment resulted in decreased CCL26 and increased IL-6 expression in the nasal and small airway cells, but this effect was not observed in the bronchial cells. IL-6 and CXCL8 were produced in varying degrees by all of the epithelial primary cells in cultures stimulated with TNF-α, LPS or IFN-γ. We showed that epithelial cells from the various levels of the respiratory tract act in a united way, responding in a similar manner to stimulation with IL-4 and IL-13, showing similar reactivity to TNF-α and LPS, and giving an almost unified response to IFN-γ pre-stimulation.

scholarly journals Analysis by Live Imaging of Effects of the Adenovirus E4orf4 Protein on Passage through Mitosis of H1299 Tumor Cells

2015 ◽  
Vol 89 (8) ◽  
pp. 4685-4689 ◽  
Author(s):  
Neera Sriskandarajah ◽  
Paola Blanchette ◽  
Thomas J. Kucharski ◽  
Jose G. Teodoro ◽  
Philip E. Branton
Keyword(s):  
Cancer Cells ◽  
Tumor Cells ◽  
Live Imaging ◽  
Primary Cells ◽  
Human Primary Cells ◽  
Normal Human

The adenovirus E4orf4 protein expressed at high levels kills cancer cells but not normal human primary cells. Previous studies suggested that disruption of processes that regulate mitosis may underlie E4orf4 toxicity. Here we have used live imaging to show that E4orf4 induces a slowed defective transit through mitosis, exhibiting a delay or often failure in cytokinesis that may account for an accumulation of G1tetraploids in the population of dying E4orf4-expressing cells.

Ultrastructural modification of cellular interactions in cancer tumor

1978 ◽  
Vol 36 (2) ◽  
pp. 318-319
Author(s):  
N. P. Dmitrieva
Keyword(s):  
Cancer Cells ◽  
Human Thyroid ◽  
Adjacent Cell ◽  
Main Role ◽  
Cellular Interactions ◽  
Electron Microscopic ◽  
Cancer Tumor ◽  
Normal Human ◽  
Characteristic Features

One of the most characteristic features of cancer cells is their ability to metastasia. It is suggested that the modifications of the structure and properties of cancer cells surfaces play the main role in this process. The present work was aimed at finding out what ultrastructural features apear in tumor in vivo which removal of individual cancer cells from the cell population can provide. For this purpose the cellular interactions in the normal human thyroid and cancer tumor of this gland electron microscopic were studied. The tissues were fixed in osmium tetroxide and were embedded in Araldite-Epon.In normal human thyroid the most common type of intercellular contacts was represented by simple junction formed by the parallelalignment of adjacent cell membranees leaving in between an intermembranes space 15-20 nm filled with electronlucid material (Fig. 1a). Sometimes in the basal part of cells dilatations of the intercellular space 40-50 nm wide were found (Fig. 1a). Here the cell surfaces may form single short microvilli.

Scanning Microscopy of Human Intestinal Explants in Organ Culture

1972 ◽  
Vol 30 ◽  
pp. 396-397
Author(s):  
Bruce Wetzel ◽  
Robert Buscho ◽  
Raphael Dolin
Keyword(s):  
Electron Microscopy ◽  
Room Temperature ◽  
Culture Conditions ◽  
Human Fetuses ◽  
Enteric Disease ◽  
Organ Cultures ◽  
Growth Of A Variety ◽  
Normal Human ◽  
Fetal Intestine ◽  
Scanning Electron

It has been reported that explants of human fetal intestine can be maintained in culture for up to 21 days in a viable condition and that these organ cultures support the growth of a variety of known viral agents responsible for enteric disease. Scanning electron microscopy (SEM) has been undertaken on several series of these explants to determine their appearance under routine culture conditions.Fresh specimens of jejunum obtained from normal human fetuses were washed, dissected into l-4mm pieces, and cultured in modified Leibowitz L-15 medium at 34° C as previously described. Serial specimens were fixed each day in 3% glutaraldehyde for 90 minutes at room temperature, rinsed, dehydrated, and dried by the CO2 critical point method in a Denton DCP-1 device. Specimens were attached to aluminum stubs with 3M transfer tape No. 465, and one sample on each stub was carefully rolled along the adhesive such that villi were broken off to expose their interiors.

Human Stapes Crura: Normal Ultrastructure. Scanning Electron Microscopic Findings

1975 ◽  
Vol 33 ◽  
pp. 528-529
Author(s):  
M.D. Graham
Keyword(s):  
Electron Microscope ◽  
Surgical Treatment ◽  
Scanning Electron Microscope ◽  
Osmic Acid ◽  
Human Cadaver ◽  
Electron Microscopic ◽  
Scanning Electron Microscopic ◽  
Normal Human ◽  
Microscopic Findings ◽  
Scanning Electron

The recent development of the scanning electron microscope has added great impetus to the study of ultrastructural details of normal human ossicles. A thorough description of the ultrastructure of the human ossicles is required in order to determine changes associated with disease processes following medical or surgical treatment.Human stapes crura were obtained at the time of surgery for clinical otosclerosis and from human cadaver material. The specimens to be examined by the scanning electron microscope were fixed immediately in the operating room in a cold phosphate buffered 2% gluteraldehyde solution, washed with Ringers, post fixed in cold 1% osmic acid and dehydrated in graded alcohol. Specimens were transferred from alcohol to a series of increasing concentrations of ethyl alcohol and amyl acetate. The tissue was then critical point dried, secured to aluminum stubs and coated with gold, approximately 150A thick on a rotating stage in a vacuum evaporator. The specimens were then studied with the Kent-Cambridge S4-10 Scanning Electron Microscope at an accelerating voltage of 20KV.

Albumin-Induced Endocytic Vacuoles in Tetrahymena Pyrijormis

1975 ◽  
Vol 33 ◽  
pp. 694-695
Author(s):  
L. J. Brenner ◽  
D. G. Osborne ◽  
B. L. Schumaker
Keyword(s):  
Electron Microscopy ◽  
Bovine Serum Albumin ◽  
Bovine Serum ◽  
Protein Concentration ◽  
Tetrahymena Pyriformis ◽  
Sequence Of Events ◽  
Cytoplasmic Bodies ◽  
Food Vacuoles ◽  
Mouse Ascites ◽  
Normal Human

Exposure of the ciliate, Tetrahymena pyriformis, strain WH6, to normal human or rabbit sera or mouse ascites fluids induces the formation of large cytoplasmic bodies. By electron microscopy these (LB) are observed to be membrane-bounded structures, generally spherical and varying in size (Fig. 1), which do not resemble the food vacuoles of cells grown in proteinaceous broth. The possibility exists that the large bodies represent endocytic vacuoles containing material concentrated from the highly nutritive proteins and lipoproteins of the sera or ascites fluids. Tetrahymena mixed with bovine serum albumin or ovalbumin solutions having about the same protein concentration (7g/100 ml) as serum form endocytic vacuoles which bear little resemblance to the serum-induced LB. The albumin-induced structures (Fig. 2) are irregular in shape, rarely spherical, and have contents which vary in density and consistency. In this paper an attempt is made to formulate the sequence of events which might occur in the formation of the albumin-induced vacuoles.

The use of 1nm silver enhanced gold probes for the detection of skin antigens

1990 ◽  
Vol 48 (3) ◽  
pp. 902-903
Author(s):  
J.P Cassella ◽  
H. Shimizu ◽  
A. Ishida-Yamamoto ◽  
R.A.J. Eady
Keyword(s):  
Type Iv Collagen ◽  
Freeze Substitution ◽  
Collagen Type Iv ◽  
Electron Microscopic ◽  
Normal Human Skin ◽  
Type Iv ◽  
Type Vii Collagen ◽  
Keratin Filaments ◽  
Normal Human ◽  
Phosphate Buffered Saline

1nm colloidal gold with silver enhancement has been used in conjunction with a low-temperature post-embedding (post-E) technique for the demonstration of skin antigens at both the light microscopic (LM) and electron microscopic (EM) levels.Keratin filaments and basement membrane zone (BMZ) associated antigens in normal human skin (NHS) were immunolabelled using antibodies against keratin 14, 10, and 1, the carboxy-terminus and collagenous portion of type VII collagen, type IV collagen and bullous pemphigoid antigen (BP-Ag).Fresh samples of NHS were cryoprotected in 15% glycerol, cryofixed in propane at -190°C, subjected to freeze substitution in methanol at -80°C and embedded in Lowicryl K11M at -60°C. Polymerisation of the resin was initiated under UVR at - 60°C for 48 hours and continued at room temperature for a further 48 hours. Semith in sections were air dried onto slides coated with 3-aminopropyltriethoxysilane. The following immunolabelling protocol was adopted: Primary antibody was applied for 2 hours at 37°C or overnight at 4°C. Following washing in Dulbecco’s phosphate buffered saline (PBSA) a biotinylated secondary antibody was applied for 2 hours at 37°C. The sections were further washed in PBSA and 1nm gold avidin was applied. Sections were finally washed in PBSA and silver enhanced.

Immunoelectron microscope detection of C-type natriuretic peptide in normal human atrial tissue

1993 ◽  
Vol 51 ◽  
pp. 388-389
Author(s):  
Chi-Ming Wei ◽  
Margaret Hukee ◽  
Christopher G.A. McGregor ◽  
John C. Burnett
Keyword(s):  
Amino Acid ◽  
Natriuretic Peptide ◽  
Vascular Tone ◽  
Cardiac Tissue ◽  
Ring Structure ◽  
Terminal Amino Acid ◽  
Amino Acid Peptide ◽  
Normal Human ◽  
Terminal Amino ◽  
The Brain

C-type natriuretic peptide (CNP) is a newly identified peptide that is structurally related to atrial (ANP) and brain natriuretic peptide (BNP). CNP exists as a 22-amino acid peptide and like ANP and BNP has a 17-amino acid ring formed by a disulfide bond. Unlike these two previously identified cardiac peptides, CNP lacks the COOH-terminal amino acid extension from the ring structure. ANP, BNP and CNP decrease cardiac preload, but unlike ANP and BNP, CNP is not natriuretic. While ANP and BNP have been localized to the heart, recent investigations have failed to detect CNP mRNA in the myocardium although small concentrations of CNP are detectable in the porcine myocardium. While originally localized to the brain, recent investigations have localized CNP to endothelial cells consistent with a paracrine role for CNP in the control of vascular tone. While CNP has been detected in cardiac tissue by radioimmunoassay, no studies have demonstrated CNP localization in normal human heart by immunoelectron microscopy.

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