scholarly journals A cellular automata to simulate the growth and death of a cell culture

2016 ◽  
Author(s):  
Giuseppe Guerrisi ◽  
Deborah Giordano ◽  
Anna Marabotti ◽  
Giancarlo Raiconi ◽  
Roberto Tagliaferri
Keyword(s):  
Cell Cycle ◽  
Cell Culture ◽  
Cell Death ◽  
Cellular Automata ◽  
Toxic Substance ◽  
Cellular Growth ◽  
Cell Counting ◽  
Physical Factors ◽  
A Cell ◽  
Toxic Factors

Motivation: The term “cell culture” is generally referred to the process by which some cells, often derived from multicellular organisms or tissues, or tumoral cell lines, are grown under controlled conditions outside of their natural environment. This system is very useful for different applications, for example to study physiological phenomena, or for the production of some useful molecules, or for testing the toxicity of some compounds. The life of the cells in culture is conditioned by many elements. Apart from physical factors such as pH and temperature, the growth of a cell culture is conditioned by its density: cells compete for the nutrients and growth factors available and die when they are exhausted. Moreover, dead cells release in the medium some toxic factors that, in their turn, can lead the surrounding cells to death. Additionally, the presence of exogenous toxic factors in the medium can induce cell death We present a cellular automata developed in order to reproduce the growth of a cell culture of a particular human cell line, Caco-2, derived from human colorectal adenocarcinoma cells. The cellular automata has been developed in order to reproduce the phenotype of Caco-2 cells, their cell cycle with all phases, and the influence of 4-nonylphenol (4-NP), an environmental pollutant, on this model system. Methods: The cellular automata developed is a grid whose dimensions reproduce a cell counting Burker chamber. Two matrices have been used to take into account, respectively, the global duration of the cellular growth and the phase of the cell cycle for each cell. Two vectors are also introduced to take into account the length of each phase and their variability range. A shuffling algorithm is used to distribute the starting cells on the chamber, then the algorithm starts by assigning a variable lag phase before reproducing the start of the cell cycle with the entering of the cells in G1 phase. All the following phases of the cell cycle are characterized by a fixed length (in minutes) + 10% variability. The cell death is described by a logarithmic function that is influenced by different factors: culture density, cellular senescence, presence of dead cells in the environment of each cell, introduction of a toxic substance. The application was developed in a stand-alone manner and has been written in Java using the OpenGL library integrated in Java. Results The application is made by an intuitive GUI to set several parameters useful for the simulation (see Figure, panel A). In order to highlight the different cell cycle phases, different colors were attributed to each phase. The cellular automata is evolving in the space and in the time reproducing the four steps of the cell cycle (G1, S, G2, M). The evolution of the simulated cell growth reproduces the phenomena present in a real Caco-2 cell culture. (Abstract truncated at 3,000 characters - the full version is available in the pdf file)

scholarly journals A cellular automata to simulate the growth and death of a cell culture

2016 ◽  
Author(s):  
Giuseppe Guerrisi ◽  
Deborah Giordano ◽  
Anna Marabotti ◽  
Giancarlo Raiconi ◽  
Roberto Tagliaferri
Keyword(s):  
Cell Cycle ◽  
Cell Culture ◽  
Cell Death ◽  
Cellular Automata ◽  
Toxic Substance ◽  
Cellular Growth ◽  
Cell Counting ◽  
Physical Factors ◽  
A Cell ◽  
Toxic Factors

Motivation: The term “cell culture” is generally referred to the process by which some cells, often derived from multicellular organisms or tissues, or tumoral cell lines, are grown under controlled conditions outside of their natural environment. This system is very useful for different applications, for example to study physiological phenomena, or for the production of some useful molecules, or for testing the toxicity of some compounds. The life of the cells in culture is conditioned by many elements. Apart from physical factors such as pH and temperature, the growth of a cell culture is conditioned by its density: cells compete for the nutrients and growth factors available and die when they are exhausted. Moreover, dead cells release in the medium some toxic factors that, in their turn, can lead the surrounding cells to death. Additionally, the presence of exogenous toxic factors in the medium can induce cell death We present a cellular automata developed in order to reproduce the growth of a cell culture of a particular human cell line, Caco-2, derived from human colorectal adenocarcinoma cells. The cellular automata has been developed in order to reproduce the phenotype of Caco-2 cells, their cell cycle with all phases, and the influence of 4-nonylphenol (4-NP), an environmental pollutant, on this model system. Methods: The cellular automata developed is a grid whose dimensions reproduce a cell counting Burker chamber. Two matrices have been used to take into account, respectively, the global duration of the cellular growth and the phase of the cell cycle for each cell. Two vectors are also introduced to take into account the length of each phase and their variability range. A shuffling algorithm is used to distribute the starting cells on the chamber, then the algorithm starts by assigning a variable lag phase before reproducing the start of the cell cycle with the entering of the cells in G1 phase. All the following phases of the cell cycle are characterized by a fixed length (in minutes) + 10% variability. The cell death is described by a logarithmic function that is influenced by different factors: culture density, cellular senescence, presence of dead cells in the environment of each cell, introduction of a toxic substance. The application was developed in a stand-alone manner and has been written in Java using the OpenGL library integrated in Java. Results The application is made by an intuitive GUI to set several parameters useful for the simulation (see Figure, panel A). In order to highlight the different cell cycle phases, different colors were attributed to each phase. The cellular automata is evolving in the space and in the time reproducing the four steps of the cell cycle (G1, S, G2, M). The evolution of the simulated cell growth reproduces the phenomena present in a real Caco-2 cell culture. (Abstract truncated at 3,000 characters - the full version is available in the pdf file)

Automated cell counting of astrocytes on patterned substrates containing aliphatic and charged properties

1995 ◽  
Vol 53 ◽  
pp. 974-975
Author(s):  
W. Shain ◽  
H. Ancin ◽  
H.C. Craighead ◽  
M. Isaacson ◽  
L. Kam ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Attachment ◽  
Culture Method ◽  
Cell Counting ◽  
Data Sets ◽  
Nuclear Staining ◽  
Double Positive ◽  
A Cell ◽  
Wafer Test ◽  
Cell Densities

Neural protheses have potential to restore nervous system functions lost by trauma or disease. Nanofabrication extends this approach to implants for stimulating and recording from single or small groups of neurons in the spinal cord and brain; however, tissue compatibility is a major limitation to their practical application. We are using a cell culture method for quantitatively measuring cell attachment to surfaces designed for nanofabricated neural prostheses.Silicon wafer test surfaces composed of 50-μm bars separated by aliphatic regions were fabricated using methods similar to a procedure described by Kleinfeld et al. Test surfaces contained either a single or double positive charge/residue. Cyanine dyes (diIC18(3)) stained the background and cell membranes (Fig 1); however, identification of individual cells at higher densities was difficult (Fig 2). Nuclear staining with acriflavine allowed discrimination of individual cells and permitted automated counting of nuclei using 3-D data sets from the confocal microscope (Fig 3). For cell attachment assays, LRM5 5 astroglial cells and astrocytes in primary cell culture were plated at increasing cell densities on test substrates, incubated for 24 hr, fixed, stained, mounted on coverslips, and imaged with a 10x objective.

scholarly journals The effects of 5α-dihydrotestosterone on the kinetics of cell proliferation in rat prostate

1974 ◽  
Vol 142 (3) ◽  
pp. 483-489 ◽  
Author(s):  
Barry Lesser ◽  
Nicholas Bruchovsky
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Cellular Growth ◽  
Velocity Sedimentation ◽  
Growth Fraction ◽  
Subcutaneous Injections ◽  
Daughter Cells ◽  
A Cell ◽  
Rat Prostate ◽  
Kinetics Of

The regenerating rat prostate was used as an experimental model to determine the effects of 5α-dihydrotestosterone on certain parameters of cell proliferation, including the duration of the phases of the cell cycle and the size of the cellular growth fraction. Rats castrated 7 days previously were treated with daily subcutaneous injections of 5α-dihydrotestosterone for 14 days; 48h after the beginning of therapy, cells in the process of DNA synthesis were labelled with a single injection of radioactive thymidine and the progress of these cells through the division cycle was observed. Cell-cycle analysis was performed by fractionating prostatic nuclei according to their position in the cell cycle by using the technique of velocity sedimentation under unit gravity. The results indicate that during regeneration the cell population undergoes 1.8 doublings with a doubling time of 40h, and that the process involves almost four rounds of cell division with a cell-generation time of 20h. The growth fraction at any time is about 0.5, and about half the daughter cells produced do not re-enter the proliferative cycle. All cells present at the start of regeneration eventually undergo at least one division during the course of regeneration, although any given cell can divide from one to four times.

scholarly journals Doxorubicin and vincristine affect undifferentiated rat spermatogonia

Reproduction ◽  
2017 ◽  
Vol 153 (6) ◽  
pp. 725-735 ◽  
Author(s):  
Hermance Beaud ◽  
Ans van Pelt ◽  
Geraldine Delbes
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Dna Repair ◽  
Cell Cycle Arrest ◽  
Excision Repair ◽  
Alkylating Agents ◽  
Dna Breaks ◽  
Cycle Arrest ◽  
A Cell ◽  
The Impact

Anticancer drugs, such as alkylating agents, can affect male fertility by targeting the DNA of proliferative spermatogonial stem cells (SSC). Therefore, to reduce such side effects, other chemotherapeutics are used. However, less is known about their potential genotoxicity on SSC. Moreover, DNA repair mechanisms in SSC are poorly understood. To model treatments deprived of alkylating agents that are commonly used in cancer treatment, we tested the impact of exposure to doxorubicin and vincristine, alone or in combination (MIX), on a rat spermatogonial cell line with SSC characteristics (GC-6spg). Vincristine alone induced a cell cycle arrest and cell death without genotoxic impact. On the other hand, doxorubicin and the MIX induced a dose-dependent cell death. More importantly, doxorubicin and the MIX induced DNA breaks, measured by the COMET assay, at a non-cytotoxic dose. To elucidate which DNA repair pathway is activated in spermatogonia after exposure to doxorubicin, we screened the expression of 75 genes implicated in DNA repair. Interestingly, all were expressed constitutively in GC-6spg, suggesting great potential to respond to genotoxic stress. Doxorubicin treatments affected the expression of 16 genes (>1.5 fold change;P < 0.05) involved in cell cycle, base/nucleotide excision repair, homologous recombination and non-homologous end joining (NHEJ). The significant increase in CDKN1A and XRCC1 suggest a cell cycle arrest and implies an alternative NHEJ pathway in response to doxorubicin-induced DNA breaks. Together, our results support the idea that undifferentiated spermatogonia have the ability to respond to DNA injury from chemotherapeutic compounds and escape DNA break accumulation.

Mutations of the fizzy locus cause metaphase arrest in Drosophila melanogaster embryos

Development ◽  
1993 ◽  
Vol 117 (1) ◽  
pp. 359-376 ◽  
Author(s):  
I.A. Dawson ◽  
S. Roth ◽  
M. Akam ◽  
S. Artavanis-Tsakonas
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Drosophila Melanogaster ◽  
Double Mutant ◽  
Nuclear Division ◽  
Normal Function ◽  
Metaphase Arrest ◽  
Nervous Systems ◽  
Dividing Cells ◽  
A Cell

We describe the effects of mutations in the fizzy gene of Drosophila melanogaster and show that fizzy mutations cause cells in mitosis to arrest at metaphase. We show that maternally supplied fizzy activity is required for normal nuclear division in the preblastoderm embryo and, during later embryogenesis, that zygotic fizzy activity is required for the development of the ventrally derived epidermis and the central and peripheral nervous systems. In fizzy embryos, dividing cells in these tissues arrest at metaphase, fail to differentiate and ultimately die. In the ventral epidermis, if cells are prevented from entering mitosis by using a string mutation, cell death is prevented and the ability to differentiate ventral epidermis is restored in fizzy; string double mutant embryos. These results demonstrate that fizzy is a cell cycle mutation and that the normal function of the fizzy gene is required for dividing cells to exit metaphase and complete mitosis.

scholarly journals Palbociclib Induces Senescence in Melanoma and Breast Cancer Cells and Leads to Additive Growth Arrest in Combination With Irradiation

2021 ◽  
Vol 11 ◽  
Author(s):  
Tina Jost ◽  
Lucie Heinzerling ◽  
Rainer Fietkau ◽  
Markus Hecht ◽  
Luitpold V. Distel
Keyword(s):  
Breast Cancer ◽  
Cell Cycle ◽  
Cell Death ◽  
Cell Cycle Arrest ◽  
Cancer Cells ◽  
Growth Arrest ◽  
Melanoma Cells ◽  
Cellular Growth ◽  
Cycle Arrest ◽  
Wide Range

IntroductionSeveral kinase inhibitors (KI) bear the potential to act as radiosensitizers. Little is known of the radiosensitizing effects of a wide range of other KI like palbociclib, which is approved in ER+/HER2- metastatic breast cancer.MethodIn our study, we used healthy donor fibroblasts and breast cancer and skin cancer cells to investigate the influence of a concomitant KI + radiation therapy. Cell death and cell cycle distribution were studied by flow cytometry after Annexin-V/7-AAD and Hoechst staining. Cellular growth arrest was studied in colony-forming assays. Furthermore, we used C12-FDG staining (senescence) and mRNA expression analysis (qPCR) to clarify cellular mechanisms.ResultsThe CDK4/6 inhibitor palbociclib induced a cell cycle arrest in the G0/G1 phase. Cellular toxicity (cell death) was only slightly increased by palbociclib and not enhanced by additional radiotherapy. As the main outcome of the colony formation assays, we found that cellular growth arrest was induced by palbociclib and improved by radiotherapy in an additive manner. Noticeably, palbociclib treatment clearly induced senescence not only in breast cancer and partly in melanoma cells, but also in healthy fibroblasts. According to these findings, the downregulation of senescence-related FOXM1 might be an involved mechanism of the senescence-induction potential of palbociclib.ConclusionThe effect on cellular growth arrest of palbociclib and radiotherapy is additive. Palbociclib induces permanent G0/G1 cell cycle arrest by inducing senescence in fibroblasts, breast cancer, and melanoma cells. Direct cell death induction is only a minor secondary mechanism of action. Concomitant KI and radiotherapy is a strategy worth studying in clinical trials.

Differences of palbociclib and ribociclib in terms of cell death pathways; a cell culture assay

2020 ◽  
Author(s):  
Ahmet Ozluk ◽  
Gulcan Bulut ◽  
Harika Atmaca ◽  
Saziye Karaca
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Cell Death ◽  
Cell Lines ◽  
Hormone Receptor ◽  
Resistance Mechanism ◽  
Cell Death Pathways ◽  
A Cell ◽  
Cell Culture Assay ◽  
Mcf 7

Abstract Differences of palbociclib and ribociclib in terms of cell death pathways; a cell culture assay The contribution of adding cyclin dependent kinase 4/6 (CDK4/6) inhibitors to hormonotherapy in relation with the survival was indicated for the treatment of metastatic breast cancer which had positive hormone receptor and negative HER2. In this study, it was planned to indicate two different CDK4/6 inhibitors (palbociclib and ribociclib) providing CDK4/6 inhibition on the cell lines (MCF-7 and BT474) having specifications of luminal-A and luminal-B, the molecular sub-types of positive hormone receptor of breast cancer, and to reveal the molecular differences and to compare cytotoxicity data and necrosis mechanisms in the cell culture experiments. It was planned to determine the differences of resistance mechanism of new combinations. MCF-7 and BT474 cell lines were handled with Palbociclib and Ribociclib individually. The possible cytotoxicity, apoptosis and authopagy effects and the levels of apoptotic proteins were examined. It was indicated that Palbociclib and Ribociclib had cytotoxic effects in both breast cancer cells. In regard of comparing from the point of intracellular pathways, it was determined that the effect of Palbociclib decreased in the increased dosage but that the effect of Ribociclib on cell indicated by means of apoptosis. It was found that palbociclib induced autophagy in the autophagy experiment conducted on the decrease of apoptotic activity with increasing doses. Even though they worked on the same pathway, Palbociclib and Ribociclib utilized different mechanisms to kill the cells and new evidences were obtained that the resistance mechanism may be different from each other according to the treatment.

scholarly journals Neuroprotective Effect of Resveratrol Against Methamphetamine-Induced Dopaminergic Apoptotic Cell Death in a Cell Culture Model of Neurotoxicity

2011 ◽  
Vol 9 (1) ◽  
pp. 49-53 ◽  
Author(s):  
Kavin Kanthasamy ◽  
Richard Gordon ◽  
Huajun Jin ◽  
Vellareddy Anantharam ◽  
Syed Ali ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Death ◽  
Apoptotic Cell ◽  
Neuroprotective Effect ◽  
Apoptotic Cell Death ◽  
Cell Culture Model ◽  
Culture Model ◽  
A Cell

scholarly journals Pulsed 3.5 GHz high power microwaves irradiation on physiological solution and their biological evaluation on human cell lines

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Pradeep Bhartiya ◽  
Sohail Mumtaz ◽  
Jun Sup Lim ◽  
Neha Kaushik ◽  
Pradeep Lamichhane ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Culture ◽  
Cell Death ◽  
Membrane Potential ◽  
Biological Evaluation ◽  
Deionized Water ◽  
Metabolic Energy ◽  
Normal Cells ◽  
Mw Irradiation ◽  
Biological Solutions

AbstractMicrowave (MW) radiation is increasingly being used for several biological applications. Many investigations have focused on understanding the potential influences of pulsed MW irradiation on biological solutions. The current study aimed to investigate the effects of 3.5 GHz pulsed MW radiation-irradiated liquid solutions on the survival of human cancer and normal cells. Different physiological solutions such as phosphate buffer saline, deionized water, and Dulbecco’s modified Eagle medium (DMEM) for cell culture growth were irradiated with pulsed MW radiation (45 shots with the energy of 1 mJ/shot). We then evaluated physiological effects such as cell viability, metabolic activity, mitochondrial membrane potential, cell cycle, and cell death in cells treated with MW-irradiated biological solutions. As MW irradiation with power density ~ 12 kW/cm2 mainly induces reactive nitrogen oxygen species in deionized water, it altered the cell cycle, membrane potential, and cell death rates in U373MG cells due to its high electric field ~ 11 kV/cm in water. Interestingly, MW-irradiated cell culture medium and phosphate-buffered saline did not alter the cellular viability and metabolic energy of cancer and normal cells without affecting the expression of genes responsible for cell death. Taken together, MW-irradiated water can alter cellular physiology noticeably, whereas irradiated media and buffered saline solutions induce negligible or irrelevant changes that do not affect cellular health.

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