scholarly journals Cytogenetic Analysis of Primary Cultures and Cell Lines: Generalities, Applications and Protocols

10.5772/34200 ◽  
2012 ◽  
Author(s):  
Sandra Milena Rondn Lagos ◽  
Nelson Enrique Rangel Jimnez
Keyword(s):  
Cell Lines ◽  
Cytogenetic Analysis ◽  
Primary Cultures

(EB) Virus: Latency and Expression in Transplanted and Cultured Human Cells

1971 ◽  
Vol 29 ◽  
pp. 368-369
Author(s):  
B. G. Uzman ◽  
M. M. Kasac ◽  
H. Saito ◽  
A. Krishan
Keyword(s):  
Cell Lines ◽  
Primary Cultures ◽  
Virus Particles ◽  
Barr Virus ◽  
Virus Latency ◽  
Virus Surveillance ◽  
Cultured Human Cells ◽  
Epstein Barr ◽  
Serial Transplantation ◽  
Tubular Arrays

In conjunction with the cultivation and transplantation of cells from human tumors by the Programs of Microbiology and Immunogenetics, virus surveillance by electron microscopy has been routinely employed. Of particular interest in this regard have been 3 cell lines cultured from lymph nodes or spleen of 2 patients with Hodgkin's disease and 1 patient with Letterer-Siwe's disease. Each of these cell lines when transplanted in Syrian hamster neonates conditioned with anti-lymphocyte serum grew as serially transplantable tumors; from such transplants of the 3 cell lines cell cultures were retrieved.Herpes type virus particles (Figs. 1, 2, 3) were found in the primary cultures of all three lines, in frozen thawed aliquots of same, and in cultures retrieved from their tumors growing by serial transplantation in hamsters. No virus was detected in sections of 25 of the serially transplanted tumors. However, in 10 such tumors there were repeated instances of tubular arrays in the cisternae of the endoplasmic reticulum (Fig. 4). On serologic examination the herpes virus was shown to be the Epstein-Barr virus.

Acute and Persistent Infection of Human Neural Cell Lines by Human Coronavirus OC43

1999 ◽  
Vol 73 (4) ◽  
pp. 3338-3350 ◽  
Author(s):  
Nathalie Arbour ◽  
Geneviève Côté ◽  
Claude Lachance ◽  
Marc Tardieu ◽  
Neil R. Cashman ◽  
...  
Keyword(s):  
Nervous System ◽  
Cell Lines ◽  
Persistent Infection ◽  
Viral Antigen ◽  
Primary Cultures ◽  
Point Mutations ◽  
Neural Cell ◽  
Persistently Infected ◽  
Human Neural Cell ◽  
Neural Cell Lines

ABSTRACT Human coronaviruses (HuCV) are recognized respiratory pathogens. Data accumulated by different laboratories suggest their neurotropic potential. For example, primary cultures of human astrocytes and microglia were shown to be susceptible to an infection by the OC43 strain of HuCV (A. Bonavia, N. Arbour, V. W. Yong, and P. J. Talbot, J. Virol. 71:800–806, 1997). We speculate that the neurotropism of HuCV will lead to persistence within the central nervous system, as was observed for murine coronaviruses. As a first step in the verification of our hypothesis, we have characterized the susceptibility of various human neural cell lines to infection by HuCV-OC43. Viral antigen, infectious virus progeny, and viral RNA were monitored during both acute and persistent infections. The astrocytoma cell lines U-87 MG, U-373 MG, and GL-15, as well as neuroblastoma SK-N-SH, neuroglioma H4, oligodendrocytic MO3.13, and the CHME-5 immortalized fetal microglial cell lines, were all susceptible to an acute infection by HuCV-OC43. Viral antigen and RNA and release of infectious virions were observed during persistent HuCV-OC43 infections (∼130 days of culture) of U-87 MG, U-373 MG, MO3.13, and H4 cell lines. Nucleotide sequences of RNA encoding the putatively hypervariable viral S1 gene fragment obtained after 130 days of culture were compared to that of initial virus input. Point mutations leading to amino acid changes were observed in all persistently infected cell lines. Moreover, an in-frame deletion was also observed in persistently infected H4 cells. Some point mutations were observed in some molecular clones but not all, suggesting evolution of the viral population and the emergence of viral quasispecies during persistent infection of H4, U-87 MG, and MO3.13 cell lines. These results are consistent with the potential persistence of HuCV-OC43 in cells of the human nervous system, accompanied by the production of infectious virions and molecular variation of viral genomic RNA.

Toxoplasma gondii invasion and replication in astrocyte primary cultures and astrocytoma cell lines: systematic review of the literature

2012 ◽  
Vol 110 (6) ◽  
pp. 2089-2094 ◽  
Author(s):  
Carla O. Contreras-Ochoa ◽  
Alfredo Lagunas-Martínez ◽  
Jaime Belkind-Gerson ◽  
Dolores Correa
Keyword(s):  
Systematic Review ◽  
Toxoplasma Gondii ◽  
Cell Lines ◽  
Primary Cultures ◽  
Review Of The Literature ◽  
Astrocytoma Cell

scholarly journals Comparative activity of pulsed or continuous estradiol exposure on gene expression and proliferation of normal and tumoral human breast cells

2002 ◽  
Vol 28 (3) ◽  
pp. 165-175 ◽  
Author(s):  
V Cavailles ◽  
A Gompel ◽  
MC Portois ◽  
S Thenot ◽  
N Mabon ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Hormone Replacement ◽  
Primary Cultures ◽  
Continuous Treatment ◽  
Luciferase Reporter ◽  
Breast Cancer Cell Lines ◽  
Human Breast ◽  
Kinetic Profile ◽  
Breast Cells

Intranasal administration of hormone replacement therapy presents an original plasma kinetic profile with transient estrogen levels giving rise to the concept of pulsed therapy. To further understand the molecular effects of this new therapy, we have compared the effects of pulsed and continuous estradiol treatments on two critical aspects of estradiol action: gene expression and cell proliferation. Cells were stimulated with estradiol as 1-h pulsed or 24-h continuous treatments at concentrations such that the 24-h exposure (concentration x time) was identical in both conditions. In MCF7 cells, the transcriptional activity of estrogen receptors (ER) on a transiently transfected responsive estrogen response element-luciferase reporter construct was shown to be drastically (approximately 10-fold) and similarly stimulated after both treatments. Moreover, the increased mRNA expression of three representative estradiol-sensitive genes (pS2, cathepsin D, progesterone receptor), evaluated by Northern blot, was identical after 1-h pulse with 7 nM estradiol or continuous treatment with 0.29 nM estradiol with the same kinetic profile over 48 h. Proliferation was quantified by a histomorphometric method on primary cultures of human normal breast cells from reduction mammoplasties and using a fluorescence DNA assay in six human breast cancer cell lines which were ER positive or negative. After a 7-day treatment period, estradiol had no effect on the proliferation of the three ER negative cell lines (BT20, MDA MB231, SK BR3) but significantly stimulated the proliferation of the normal cells and of the three tumoral hormone-sensitive cell lines (MCF7, T47D, ZR 75-1); both hormone treatments producing the same increases in cell growth. In conclusion, we have shown that the genomic or proliferative effects of estradiol were identical with pulsed or continuous treatments, thus indicating that estrogenic effects are not strictly related to concentrations but rather to total hormone exposure.

Glucose metabolism by adult hepatocytes in primary culture and by cell lines from rat liver

1978 ◽  
Vol 234 (3) ◽  
pp. C122-C130 ◽  
Author(s):  
D. M. Bissell ◽  
G. A. Levine ◽  
M. J. Bissell
Keyword(s):  
Glucose Metabolism ◽  
Primary Culture ◽  
Cell Lines ◽  
Rat Liver ◽  
Liver Cell ◽  
Time Course ◽  
Primary Cultures ◽  
Functional Change ◽  
Permanent Cell ◽  
Rat Hepatoma

The metabolic fate of [U-14C]glucose has been examined in detail in adult rat hepatocytes in primary monolayer culture, as well as in two permanent cell lines--Buffalo rat liver (BRL) and transplantable rat hepatoma (HTC) cells-derived from normal rat liver and from rat hepatoma, respectively. Under defined conditions of incubation, at a glucose concentration of 5.5 mM, the three types of cultured liver cells exhibited pronounced differences in glucose metabolism. Primary cultures, like the intact liver, differed from the cell lines in consuming relatively small amounts of glucose and converting approximately 50% of the total metabolized glucose to lactate. By contrast, the permantent cell lines consumed glucose at a 40-fold greater rate than did primary cultures, converting 80--90% of the carbohydrate to lactate. Oxidative metabolism of glucose carbon also differed among the three types of liver culture. Of the total [U-14C]glucose consumed, primary cultures converted approximately 30% to labeled CO2 per hour, whereas the liver cell lines converted 5--10%. Finally, glucose metabolism in primary culture exhibited adaptation as hepatocytes aged in culture, shifting progressively toward the pattern exhibited by the permanent cell lines. This change occurred over a time course similar to that for other kinds of functional change in hepatocytes in primary culture and thus may be relevant to the general problem of phenotypic alteration in liver cell culture.

scholarly journals Primary Cultures and Cell Lines for In Vitro Modeling of the Human Adrenal Cortex

2021 ◽  
Vol 253 (4) ◽  
pp. 217-232
Author(s):  
Kazutaka Nanba ◽  
Amy R. Blinder ◽  
William E. Rainey
Keyword(s):  
Adrenal Cortex ◽  
Cell Lines ◽  
Primary Cultures

Cytogenetic analysis of melanoma cell lines

2003 ◽  
Vol 142 (2) ◽  
pp. 87-91 ◽  
Author(s):  
Michal Lotem ◽  
Orly Yehuda-Gafni ◽  
Eliza Butnaryu ◽  
Olga Drize ◽  
Tamar Peretz ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Cytogenetic Analysis ◽  
Melanoma Cell Lines

scholarly journals Interleukin-HP1, a T cell-derived hybridoma growth factor that supports the in vitro growth of murine plasmacytomas.

1987 ◽  
Vol 165 (3) ◽  
pp. 641-649 ◽  
Author(s):  
J Van Snick ◽  
A Vink ◽  
S Cayphas ◽  
C Uyttenhove
Keyword(s):  
T Cell ◽  
Growth Factor ◽  
B Cell ◽  
Cell Lines ◽  
Primary Cultures ◽  
Gel Filtration ◽  
Reversed Phase ◽  
Hybridoma Growth ◽  
Cell Supernatant

We have recently described the purification and NH2-terminal amino acid sequence of a T cell-derived hybridoma growth factor that was provisionally designated interleukin-HP1 (IL-HP1). Here we report that a T cell supernatant containing high titers of this hybridoma growth factor considerably facilitated the establishment of primary cultures of murine plasmacytomas. Most plasmacytoma cell lines derived from such cultures remained permanently dependent on IL-HP1-containing T cell supernatant for both survival and growth in vitro. These cell lines, however, retained their ability to form tumors in irradiated pristane-treated mice. Analytical fractionation of a T cell supernatant rich in IL-HP1 by either gel filtration, isoelectric focusing, or reversed-phase HPLC revealed the existence of only one plasmacytoma growth factor activity that strictly copurified with IL-HP1, strongly suggesting the identity of both factors. This conclusion was further supported by the finding that IL-HP1 purified to homogeneity supported the growth of both B cell hybridomas and plasmacytomas. For half-maximal growth, plasmacytomas, however, required a concentration of IL-HP1 of approximately 30 pM, which is approximately 200 times higher than that required by B cell hybridomas. A clear difference in the specificity of IL-HP1 and B cell stimulatory factor 1 (BSF-1) was demonstrated by the finding that IL-HP1-dependent plasmacytomas did not survive in the presence of BSF-1, whereas helper T cell lines that proliferated in the presence of BSF-1 failed to respond to IL-HP1.

Liver-specific RNA metabolism in hepatoma cells: variations in transcription rates and mRNA levels

1985 ◽  
Vol 5 (10) ◽  
pp. 2633-2641
Author(s):  
D F Clayton ◽  
M Weiss ◽  
J E Darnell
Keyword(s):  
Cell Lines ◽  
Transcriptional Control ◽  
Hepatoma Cell ◽  
Primary Cultures ◽  
Normal Liver ◽  
Liver Cells ◽  
Hepatoma Cell Line ◽  
Mrna Levels ◽  
Tissue Specific ◽  
Specific Mrnas

The transcription rate and abundance of several liver-specific mRNAs as well as mRNAs common to many cell types were compared in a series of rodent hepatoma cell lines, normal liver cells, and primary hepatocyte cultures. The rat hepatoma cell line, Fao, which displays a liver-specific phenotype, contained eight of eight liver-specific mRNAs examined. However, the transcription rates of most liver-specific mRNAs were found to be low (1 to 30%) compared with normal liver in this and other differentiated cell lines. This low rate is similar to the transcription rates of liver-specific mRNA sequences measured in primary cultures of hepatocytes. Several variant cell lines that had lost differentiated traits contained few or none of the liver-specific mRNAs; clonal descendents which had regained differentiated function regained the tissue-specific mRNAs as a group, but at various concentrations. Because all of the changes observed in mRNA levels were not accompanied by parallel changes in transcription of the same sequences, differential posttranscriptional stabilization of the liver-specific mRNAs must also occur in the different cell lines. These results qualify the utility of cultured cell lines in the study of tissue-specific transcriptional control, but raise the possibility that posttranscriptional mechanisms act in cooperation with transcriptional controls to bring the level of tissue-specific mRNAs closer to those found in liver cells.

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