Root tip cell cycle synchronization and metaphase-chromosome isolation suitable for flow sorting in common wheat (Triticum aestivum L.)

Genome ◽  
1997 ◽  
Vol 40 (5) ◽  
pp. 633-638 ◽  
Author(s):  
Jai-Heon Lee ◽  
K. Arumuganathan ◽  
Y. Yen ◽  
S. Kaeppler ◽  
H. Kaeppler ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Common Wheat ◽  
Univariate Analysis ◽  
Flow Cytometric Analysis ◽  
Root Tip ◽  
Bivariate Analysis ◽  
Root Tips ◽  
Metaphase Chromosomes ◽  
Cell Cycle Synchronization ◽  
Flow Cytometric

An efficient procedure for cell-cycle synchronization in meristematic root tips was achieved in common wheat. Treatment parameters for synchronizing the cell cycle of root tip meristem cells, such as time-course and applied concentrations of various chemicals, were systematically tested and optimized by flow cytometric analysis of isolated nuclei. High mitotic indices (69.5% in the root tip meristematic area) were routinely obtained by treating germinating seeds with 1.25 mM hydroxyurea for 16 h, followed by incubation in a hydroxyurea-free solution for 2 h, and treatment with 1 μM trifluralin for 4 h. Uniform seed germination prior to treatment is very important for achieving consistently high metaphase indices in the root tips. Large numbers of metaphase chromosomes, suitable for flow cytometric analysis and sorting, were isolated from synchronized root tip cells. Flow sorted wheat chromosomes, via univariate and bivariate analysis, showed four major chromosome peaks. Each discrete peak may represent wheat chromosome types with similar DNA content. Bivariate flow karyotyping based on AT and GC content did not improve the separation of wheat chromosomes.Key words: flow cytometry, trifluralin, hydroxyurea, univariate analysis, bivariate analysis.

Cell synchronization and isolation of metaphase chromosomes from maize (Zea mays L.) root tips for flow cytometric analysis and sorting

Genome ◽  
1996 ◽  
Vol 39 (4) ◽  
pp. 697-703 ◽  
Author(s):  
Jai-Heon Lee ◽  
K. Arumuganathan ◽  
S. M. Kaeppler ◽  
C. M. Papa ◽  
H. F. Kaeppler
Keyword(s):  
Cell Cycle ◽  
Flow Cytometric Analysis ◽  
Maize Root ◽  
Root Tip ◽  
Root Tips ◽  
Chromosome Sorting ◽  
Synthesis Inhibitor ◽  
Metaphase Chromosomes ◽  
Flow Karyotype ◽  
Flow Cytometric

Accumulation of cells containing metaphase chromosomes is an important step in cytological analyses and chromosome sorting procedures. The goal of this research was to optimize treatment parameters to synchronize the cell cycle of maize root tip meristem cells. Levels of hydroxyurea, a DNA synthesis inhibitor, were assessed for their utility in accumulating cells at the G1 phase of the cell cycle. Trifluralin, amiprophos-methyl, and colchicine were used to accumulate cells containing metaphase chromosomes upon release from hydroxyurea inhibition. Optimal mitotic indices were achieved by treating seedlings with 5 mM hydroxyurea for 18 h, incubating for 1 h without chemical treatment to release the hydroxyurea block, and then treating emerging roots with 1 μM trifluralin for 4 h. The mitotic index of synchronized maize root tips was over 70%. Uniformity of synchronization depended upon selection of seeds with emerging radicles that were similar in length at the time of treatment. Suspensions of intact chromosomes were prepared by a simple slicing procedure. The chromosome preparations were found to be suitable for flow cytometric characterization and sorting. Chromosome peaks of the observed flow karyotype resembled the predicted flow karyotype calculated on the basis of maize chromosome size. Key words : flow karyotype, hydroxyurea, plant chromosome sorting, trifluralin.

New thiazacridine agents: Synthesis, physical and chemical characterization, and in vitro anticancer evaluation

2016 ◽  
Vol 36 (10) ◽  
pp. 1059-1070 ◽  
Author(s):  
MBO Chagas ◽  
NCC Cordeiro ◽  
KMR Marques ◽  
MG Rocha Pitta ◽  
MJBM Rêgo ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cytotoxic Activity ◽  
Cell Cycle Progression ◽  
Antitumor Agents ◽  
Flow Cytometric Analysis ◽  
Cell Lineage ◽  
Chemical Characterization ◽  
Flow Cytometric ◽  
Physical And Chemical

A series of new thiazacridine agents were synthesized and evaluated as antitumor agents, in terms of not only their cytotoxicity but also their selectivity. The cytotoxicity assay confirmed that all compounds showed cytotoxic activity and selectivity. The new compound, 3-acridin-9-ylmethyl-5-(5-bromo-1 H-indol-3-ylmethylene)-thiazolidine-2,4-dione (LPSF/AA29 – 7a), proved to be the most promising compound as it presents lower half-maximal inhibitory concentration (IC50) values (ranging from 0.25 to 68.03 µM) depending on cell lineage. In HepG2 cells, the lowest IC50 value was exhibited by 3-acridin-9-ylmethyl-5-(4-piperidin-1-yl-benzylidene)-thiazolidine-2,4-dione (LPSF/AA36 – 7b; 46.95 µM). None of the synthesized compounds showed cytotoxic activity against normal cells (IC50 > 100 µM). The mechanism of death induction and cell cycle effects was also evaluated. Flow cytometric analysis revealed that the compounds LPSF/AA29 – 7a and LPSF/AA36 – 7b significantly increased the percentage of apoptotic cells and induced G2/M arrest in the cell cycle progression. Therefore, these new thiazacridine derivatives constitute promising antitumor agents whose cytotoxicity and selectivity properties indicate they have potential to contribute to or serve as a basis for the development of new cancer drugs in the future.

scholarly journals Flow Cytometric Analysis of the Influence of Myosmine on the Cell Cycle

2011 ◽  
Vol 84 (3) ◽  
pp. 355-359 ◽  
Author(s):  
Borislava I. Boteva ◽  
Rada M. Mateva ◽  
Ivan A. Iliev ◽  
Galina A. Gorneva
Keyword(s):  
Cell Cycle ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric

Rapid Flow Cytometric Analysis of the Cell Cycle in Intact Plant Tissues

Science ◽  
1983 ◽  
Vol 220 (4601) ◽  
pp. 1049-1051 ◽  
Author(s):  
D. W. GALBRAITH ◽  
K. R. HARKINS ◽  
J. M. MADDOX ◽  
N. M. AYRES ◽  
D. P. SHARMA ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometric Analysis ◽  
Intact Plant ◽  
Plant Tissues ◽  
Flow Cytometric ◽  
Rapid Flow

scholarly journals EXTH-56. MEDULLOBLASTOMAS RESPOND TO CDK4/6 INHIBITION WITH NANOPARTICLE-DELIVERED PALBOCICLIB WITH ALTERED S-PHASE PROGRESSION

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi94-vi94
Author(s):  
Taylor Dismuke ◽  
Chaemin Lim ◽  
Timothy Gershon
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Progression ◽  
Flow Cytometric Analysis ◽  
Pediatric Brain Tumors ◽  
S Phase ◽  
Flow Cytometric ◽  
Phase Progression ◽  
Reduced Rate ◽  
G1 Cells ◽  
Pediatric Brain

Abstract CDK4/6 inhibition is a promising therapy for medulloblastoma, one of the most common malignant pediatric brain tumors. To improve pharmacokinetics, we developed a polyoxazoline nanoparticle-encapsulated formulation of the FDA-approved CDK4/6 inhibitor palbociclib (POx-palbo). We then administered POx-palbo to transgenic medulloblastoma-prone GFAP-Cre/SmoM2 mice, to determine the efficacy and mechanisms of action and resistance. We found that POx-palbo slowed tumor progression, but consistently failed to be curative. Further analysis showed that while CDK4/6 inhibition acutely blocked G1 cells from re-entering the cell cycle, this effect wore off within hours of drug administration. However, flow cytometric analysis of EdU uptake hours after palbociclib demonstrated aberrant S-phase with reduced rate of DNA synthesis. This POx-palbociclib-induced alteration of S-phase progression seems to remain true at later time points even when we observed that palbociclib G1/S inhibition began to decrease. Based on these data, we propose that the combinational therapy of POx-palbociclib and S-phase targeting agents will further improve treatment. Faulty tumor cell cycle progression in the presence of Pox-palbociclib may give increased window to target the S-phase for irreversible cell-cycle exit.

Flow cytometric analysis of the cell-cycle distribution of spleen lymphocytes isolated from Fischer 344 rats exposed to ethyl nitrosourea

1993 ◽  
Vol 9 (1) ◽  
pp. 77-83 ◽  
Author(s):  
Suzanne M. Morris ◽  
Olen E. Domon ◽  
Lynda J. McGarrity ◽  
Anane Aidoo ◽  
Ralph L. Kodell ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometric Analysis ◽  
Cell Cycle Distribution ◽  
Fischer 344 Rats ◽  
Fischer 344 ◽  
Flow Cytometric ◽  
Spleen Lymphocytes
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