scholarly journals Adaption of FMDV Asia-1 to Suspension Culture: Cell Resistance Is Overcome by Virus Capsid Alterations

Viruses ◽  
2017 ◽  
Vol 9 (8) ◽  
pp. 231 ◽  
Author(s):  
Veronika Dill ◽  
Bernd Hoffmann ◽  
Aline Zimmer ◽  
Martin Beer ◽  
Michael Eschbaumer
Author(s):  
J. G. Holshek ◽  
R. E. Akins

Acetonitrile has been shown to be a better dehydrating agent than ethanol for whole tissue prepared for TEM studies. In this paper we show that it is also a better choice for dehydration of cell cultures prepared for SEM studies. The procedures used for the isolation and culture of cardiac cells in the present study have been outlined in detail by Schroedl and Hartzell (1983) and Freerksen, et al. (1984). Cells were cultured in suspension using HARV-Bioreactors (Synthecon, Inc., Friendswood, TX) for 24 hours to allow attachment and then fed each subsequent 48 hours. After a total of six days in suspension culture, cell/bead clusters were collected and washed with PBS.Primary fixation was done at 4°C, all other steps were performed at room temperature. The buffer used in all cases was 0.1M cacodylate, pH 7.3. The cell/bead cultures were processed according to the following protocol. All of the beads were placed in a 1.5 ml microfuge tube and fixed in 2% buffered glutaraldehyde overnight followed by two 5 min washes in buffer, postfixation in 1% buffered OsO4 for 90 min and three washes in buffer. One half of the beads were then placed in another microfuge tube and dehydrated in acetonitrile, while the remaining beads were dehydrated in ethanol.


2006 ◽  
Vol 61 (3-4) ◽  
pp. 267-272 ◽  
Author(s):  
Eiichiro Fukusaki ◽  
Kanokwan Jumtee ◽  
Takeshi Bamba ◽  
Takehiro Yamaji ◽  
Akio Kobayashi

Cell suspension cultures are now recognized as important model materials for plant bioscience and biotechnology. Very few studies of metabolic comparisons between cell cultures and original plants have been reported, even though the biological identity of cultured cells with the normally grown plant is of great importance. In this study, a comparison of the metabolome for primary metabolites extracted from the leaves of Arabidopsis thaliana and cultured cells from an Arabidopsis suspension culture (cell line T87) was performed. The results suggest that although cell suspension cultures and Arabidopsis leaves showed similarities in the common primary metabolite profile, nonetheless, moderate differences in quantitative profile were revealed.


Blood ◽  
1992 ◽  
Vol 80 (4) ◽  
pp. 1026-1032 ◽  
Author(s):  
SJ Pirruccello ◽  
JD Jackson ◽  
MS Lang ◽  
J DeBoer ◽  
S Mann ◽  
...  

Abstract OMA-AML-1 was established from a patient with acute myelomonocytic (M4) leukemia at fifth relapse when blasts were greater than 85% CD34+, CD15- . Leukemic cells were established in suspension culture and independently grown as subcutaneous tumors in SCID mice. Cells growing in suspension culture underwent differentiation by phenotypic and morphologic criteria. In contrast, cells grown as subcutaneous solid tumors in SCID mice maintained progenitor cell characteristics with high-density CD34 expression and lack of morphologic differentiation. A tendency toward differentiation to CD15+, CD34- cells in vitro and self- renewal of CD34+, CD15- cells in vivo was consistently demonstrated regardless of whether cells were initially grown in vitro or in vivo. The cell line maintains both a CD34+, CD15- progentitor cell pool and a non-overlapping, CD15+, CD34- differentiating cell compartment after more than 1 year in continuous culture. Cell cycle analysis and cloning experiments were consistent with terminal differentiation occurring in the CD15+, CD34- population. The cell line shows concentration- dependent proliferative responses to interleukin (IL)-3, granulocyte- macrophage colony-stimulating factor (GM-CSF), and IL-6, but not to granulocyte CSF (G-CSF). OMA-AML-1 appears to mimic several features of normal myeloid hematopoiesis and should prove useful for the study of normal and malignant myeloid differentiation.


Blood ◽  
1992 ◽  
Vol 80 (4) ◽  
pp. 1026-1032
Author(s):  
SJ Pirruccello ◽  
JD Jackson ◽  
MS Lang ◽  
J DeBoer ◽  
S Mann ◽  
...  

OMA-AML-1 was established from a patient with acute myelomonocytic (M4) leukemia at fifth relapse when blasts were greater than 85% CD34+, CD15- . Leukemic cells were established in suspension culture and independently grown as subcutaneous tumors in SCID mice. Cells growing in suspension culture underwent differentiation by phenotypic and morphologic criteria. In contrast, cells grown as subcutaneous solid tumors in SCID mice maintained progenitor cell characteristics with high-density CD34 expression and lack of morphologic differentiation. A tendency toward differentiation to CD15+, CD34- cells in vitro and self- renewal of CD34+, CD15- cells in vivo was consistently demonstrated regardless of whether cells were initially grown in vitro or in vivo. The cell line maintains both a CD34+, CD15- progentitor cell pool and a non-overlapping, CD15+, CD34- differentiating cell compartment after more than 1 year in continuous culture. Cell cycle analysis and cloning experiments were consistent with terminal differentiation occurring in the CD15+, CD34- population. The cell line shows concentration- dependent proliferative responses to interleukin (IL)-3, granulocyte- macrophage colony-stimulating factor (GM-CSF), and IL-6, but not to granulocyte CSF (G-CSF). OMA-AML-1 appears to mimic several features of normal myeloid hematopoiesis and should prove useful for the study of normal and malignant myeloid differentiation.


Author(s):  
R. Stephens ◽  
G. Schidlovsky ◽  
S. Kuzmic ◽  
P. Gaudreau

The usual method of scraping or trypsinization to detach tissue culture cell sheets from their glass substrate for further pelletization and processing for electron microscopy introduces objectionable morphological alterations. It is also impossible under these conditions to study a particular area or individual cell which have been preselected by light microscopy in the living state.Several schemes which obviate centrifugation and allow the embedding of nondetached tissue culture cells have been proposed. However, they all preserve only a small part of the cell sheet and make use of inverted gelatin capsules which are in this case difficult to handle.We have evolved and used over a period of several years a technique which allows the embedding of a complete cell sheet growing at the inner surface of a tissue culture roller tube. Observation of the same cell by light microscopy in the living and embedded states followed by electron microscopy is performed conveniently.


Author(s):  
Jane K. Rosenthal ◽  
Dianne L. Atkins ◽  
William J. Marvin ◽  
Penny A. Krumm

To comprehend structural changes in cardiac myocytes accompanying adrenergic innervation, it is essential that a three dimensional analysis be performed. To date, biological studies which utilize stereological methods have been limited to cells in tissue and in organs. Our laboratory has utilized current stereological techniques for measuring absolute volumes of individual myocytes in primary culture. Cell volumes are calculated for two distinct groups of cells at 96 hours in culture: isolated myocytes and myocytes innervated with adrenergic neurons (Figure 1).Cardiac myocytes are cultured from the ventricular apices of newborn rats. Cells are plated directly onto tissue culture dishes with or without preplated explants from the paravertebral thoracolumbar sympathetic chain. On day four cultures are photographed and marked for one-to-one cell location. Following conventional fixation and embeddment in eponate-12, the cells are relocated and mounted for microtomy. The cells are completely sectioned at 120nm in their parallel orientation to the surface of the dish (Figure 2). Serial sections are collected on formvar coated slotted grids and are recorded in sequence.


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