Accurate non-invasive image-based cytotoxicity assays for cultured cells

The advanced oxidation process (AOP) is used to increase the treatment efficiency of effluents however, it is necessary to compare the toxicity of treated and untreated effluents to evaluate if the decontamination process does not cause any biological harm. Cultured cells have been previously used to assess the genotoxic and cytotoxic potential of various compounds. Hence, the aim of this work was to assess the applicability of cytotoxicity assays to evaluate the toxicity related to the AOP treatment. Samples of an industrial effluent were collected after their treatment by a conventional method. Cytotoxicity of standard and AOP treated effluents was assessed in CRIB and HEp-2 cell line using the MTT and neutral red assays. We observed decrease at cell viability in the both assays (50% MTT and 13% NRU) when cells were exposed to the AOP treatment in the highest concentration. Thus, cytotoxic assays in cultured cells can be explored as an useful method to evaluate toxicity as well as to optimize effluents treatment process.

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Characterization and simulation of peristaltic micropumps

Journal of Sensors and Sensor Systems ◽

10.5194/jsss-2-165-2013 ◽

2013 ◽

Vol 2 (2) ◽

pp. 165-169 ◽

Cited By ~ 10

Author(s):

M. Busek ◽

M. Nötzel ◽

C. Polk ◽

F. Sonntag

Keyword(s):

Cell Culture ◽

Culture System ◽

Cultured Cells ◽

Applied Pressure ◽

Cell Culture System ◽

Heart Activity ◽

Non Invasive ◽

Micro Piv ◽

Piv Method ◽

A Cell

Abstract. The aim of the work is to find an analytical model of a pneumatic micropump which was integrated into a cell-culture system. This allows the estimation of peak velocities and wall-shear stress influencing the cultured cells in our multi-organ-chip (MOC) with respect to the applied pressure and the geometric properties of the pump. By adjusting those parameters, one can imitate physiological or pathological heart activity. The calculated flow within the MOC was compared to experimental results obtained via the non-invasive micro-PIV method.

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Cells/colony motion of oral keratinocytes determined by non-invasive and quantitative measurement using optical flow predicts epithelial regenerative capacity

Scientific Reports ◽

10.1038/s41598-021-89073-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Emi Hoshikawa ◽

Taisuke Sato ◽

Kenta Haga ◽

Ayako Suzuki ◽

Ryota Kobayashi ◽

...

Keyword(s):

Optical Flow ◽

Cultured Cells ◽

Primary Cultures ◽

Culture Conditions ◽

Oral Keratinocytes ◽

Regenerative Capacity ◽

Non Invasive ◽

Quantitative Measurements ◽

Full Screen ◽

Quantitative Manner

AbstractCells/colony motion determined by non-invasive, quantitative measurements using the optical flow (OF) algorithm can indicate the oral keratinocyte proliferative capacity in early-phase primary cultures. This study aimed to determine a threshold for the cells/colony motion index to detect substandard cell populations in a subsequent subculture before manufacturing a tissue-engineered oral mucosa graft and to investigate the correlation with the epithelial regenerative capacity. The distinctive proliferating pattern of first-passage [passage 1 (p1)] cells reveals the motion of p1 cells/colonies, which can be measured in a non-invasive, quantitative manner using OF with fewer full-screen imaging analyses and cell segmentations. Our results demonstrate that the motion index lower than 40 μm/h reflects cellular damages by experimental metabolic challenges although this value shall only apply in case of our culture system. Nonetheless, the motion index can be used as the threshold to determine the quality of cultured cells while it may be affected by any different culture conditions. Because the p1 cells/colony motion index is correlated with epithelial regenerative capacity, it is a reliable index for quality control of oral keratinocytes.

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The Influence of Hypoxia and pH on Bioluminescence Imaging of Luciferase-Transfected Tumor Cells and Xenografts

International Journal of Molecular Imaging ◽

10.1155/2013/287697 ◽

2013 ◽

Vol 2013 ◽

pp. 1-9 ◽

Cited By ~ 14

Author(s):

Ashraf A. Khalil ◽

Mark J. Jameson ◽

William C. Broaddus ◽

Peck Sun Lin ◽

Seth M. Dever ◽

...

Keyword(s):

Animal Models ◽

Quantitative Assessment ◽

Chronic Hypoxia ◽

Bioluminescence Imaging ◽

Cultured Cells ◽

Xenograft Model ◽

Living Animal ◽

Non Invasive

Bioluminescence imaging (BLI) is a relatively new noninvasive technology used for quantitative assessment of tumor growth and therapeutic effect in living animal models. BLI involves the generation of light by luciferase-expressing cells following administration of the substrate luciferin in the presence of oxygen and ATP. In the present study, the effects of hypoxia, hypoperfusion, and pH on BLI signal (BLS) intensity were evaluated in vitro using cultured cells and in vivo using a xenograft model in nude mice. The intensity of the BLS was significantly reduced in the presence of acute and chronic hypoxia. Changes in cell density, viability, and pH also affected BLS. Although BLI is a convenient non-invasive tool for tumor assessment, these factors should be considered when interpreting BLS intensity, especially in solid tumors that could be hypoxic due to rapid growth, inadequate blood supply, and/or treatment.

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Resonance Raman Probes for Organelle-Specific Labeling in Live Cells

Scientific Reports ◽

10.1038/srep28483 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 11

Author(s):

Andrey N. Kuzmin ◽

Artem Pliss ◽

Chang-Keun Lim ◽

Jeongyun Heo ◽

Sehoon Kim ◽

...

Keyword(s):

Cultured Cells ◽

Red Light ◽

Resonance Raman ◽

Raman Microspectroscopy ◽

Molecular Probes ◽

Live Cells ◽

High Signal ◽

Optical Window ◽

Non Invasive ◽

Raman Probe

Abstract Raman microspectroscopy provides for high-resolution non-invasive molecular analysis of biological samples and has a breakthrough potential for dissection of cellular molecular composition at a single organelle level. However, the potential of Raman microspectroscopy can be fully realized only when novel types of molecular probes distinguishable in the Raman spectroscopy modality are developed for labeling of specific cellular domains to guide spectrochemical spatial imaging. Here we report on the design of a next generation Raman probe, based on BlackBerry Quencher 650 compound, which provides unprecedentedly high signal intensity through the Resonance Raman (RR) enhancement mechanism. Remarkably, RR enhancement occurs with low-toxic red light, which is close to maximum transparency in the biological optical window. The utility of proposed RR probes was validated for targeting lysosomes in live cultured cells, which enabled identification and subsequent monitoring of dynamic changes in this organelle by Raman imaging.

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Isolation and culture of epithelial cells from stored buffalo semen and their use for the production of cloned embryos

Reproduction Fertility and Development ◽

10.1071/rd18356 ◽

2019 ◽

Vol 31 (10) ◽

pp. 1581 ◽

Cited By ~ 1

Author(s):

Monika Saini ◽

Naresh L. Selokar ◽

Rasika Rajendran ◽

Dharmendra Kumar ◽

Pradeep Kumar ◽

...

Keyword(s):

Epithelial Cell ◽

Epithelial Cells ◽

Cultured Cells ◽

Cell Attachment ◽

Somatic Cells ◽

Cleavage Rate ◽

Cytokeratin 18 ◽

Non Invasive ◽

Culture Characteristics ◽

Buffalo Semen

The aim of the present study was to isolate somatic cells from semen, a non-invasive source of donor somatic cells, for somatic cell nuclear transfer (SCNT) experiments. The study had two parts: (1) isolation and culture of somatic cells from semen, which was stored at 4°C; and (2) investigating the SCNT competence of semen-derived somatic cells. We successfully cultured somatic cells from freshly ejaculated semen, which was stored for different times (0, 4, 12, 24, 72 and 144h after semen collection) at 4°C, using a Percoll gradient method. Up to 24h storage, 100% cell attachment rates were observed; cell attachment rates of 66% were observed for the 72 and 144h storage groups. The attached cells observed in all groups examined were proliferated (100%). Cultured cells exhibited epithelial cell morphology and culture characteristics, which was further confirmed by positive expression of cytokeratin 18, an epithelial cell-type marker. We compared the SCNT competence of semen-derived epithelial cells and skin-derived fibroblasts. The cleavage rate, blastocyst production rate, total number of cells in blastocysts and the apoptotic index of blastocysts were similar for embryos produced from semen-derived epithelial cells and skin-derived fibroblasts, indicating that semen-derived epithelial cells can serve as donors for SCNT experiments. In conclusion, we demonstrate a method to culture epithelial cells from stored semen, which can be used to produce cloned embryos of breeding bulls, including remote bulls.

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Ethosome: a novel vesicular carrier for transdermal drug delivery

Journal of Drug Delivery and Therapeutics ◽

10.22270/jddt.v8i6.2028 ◽

2018 ◽

Vol 8 (6) ◽

pp. 318-326 ◽

Cited By ~ 1

Author(s):

Saquib Raza Zahid ◽

Neeraj Upmanyu ◽

Surendra Dangi ◽

Sudhir Kumar Ray ◽

Prabhat Jain ◽

...

Keyword(s):

Drug Delivery ◽

Cultured Cells ◽

Systemic Circulation ◽

Bioactive Molecules ◽

Biomedical Sciences ◽

Double Blind ◽

Non Invasive ◽

The Past ◽

Randomized Clinical Study

Delivery across skin is striking due to its easy convenience. However, drug delivery across skin is still a confront in biomedical sciences. Over the past few decades, various successful narrative devices and techniques have emerged to optimize drug delivery across skin whose barricading behaviour constricts entry of most of the therapeutic agents. Ethosomes are non-invasive delivery transporter that enables drugs to reach the deep skin layers and/or the systemic circulation. Although ethosomal systems are theoretically sophisticated, they are characterized by simplicity in their preparation, efficacy and safety. A combination that can highly inflate their application. Ethosomes are soft, malleable vesicles adapted for enhanced delivery of active agents. This article reviews work carried out method of preparation, application and characterization of ethosomal systems. Because of their exceptional structure, ethosomes are able to encapsulate and deliver through the skin highly lipophilic molecules such as testosterone, cannabinoids and minoxidil as well as cationic drugs such as trihexyphenidil and propranolol. Results obtained in a double-blind two-armed randomized clinical study showed that treatment with the ethosomal acyclovir formulation appreciably improved all the evaluated parameters. In further work, the ethosomal expertise was broadened to introduce agents into cultured cells and microorganisms. Enhanced delivery of bioactive molecules through the skin and cellular membranes by means of an ethosomal transporter opens numerous confronts and prospects for the research and future development of novel improved therapies. Keywords: Ethosomes, Skin layers, Characterization

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Cell/Colony Motion of Oral Keratinocytes Determined by Non-Invasive and Quantitative Measurement Using Optical Flow Predicts Epithelial Regenerative Capacity

10.21203/rs.3.rs-236508/v1 ◽

2021 ◽

Author(s):

Emi Hoshikawa ◽

Taisuke Sato ◽

Kenta Haga ◽

Ayako Suzuki ◽

Ryota Kobayashi ◽

...

Keyword(s):

Optical Flow ◽

Cultured Cells ◽

Primary Cultures ◽

Oral Keratinocytes ◽

Regenerative Capacity ◽

Non Invasive ◽

Quantitative Measurements ◽

Full Screen ◽

Quantitative Manner ◽

Cell Colony

Abstract Cell/colony motion determined by non-invasive, quantitative measurements using the optical flow (OF) algorithm can indicate the oral keratinocyte proliferative capacity in early-phase primary cultures. This study aimed to determine a threshold for the cell/colony motion index to detect substandard cell populations in a subsequent subculture before manufacturing a tissue-engineered oral mucosa graft and to investigate the correlation with the epithelial regenerative capacity. The distinctive proliferating pattern of first-passage (passage 1 (p1)) cells reveals the motion of p1 cells/colonies, which can be measured in a non-invasive, quantitative manner using OF with fewer full-screen imaging analyses and cell segmentations. Our results demonstrate that the motion index lower than 40 μm/hour reflects cellular damages by experimental metabolic challenges and can be used as the threshold to determine the quality of cultured cells. Because the p1 cell/colony motion index is correlated with epithelial regenerative capacity, it is a reliable index for quality control of oral keratinocytes.

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Ultrastructural Correlations between Clonal Human Tumor Colonies and in Vivo Neoplasms

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100053711 ◽

1982 ◽

Vol 40 ◽

pp. 210-211

Author(s):

M.J. Murphy ◽

R.R. Price ◽

J.C. Sloman

Keyword(s):

Cultured Cells ◽

Human Tumor ◽

Cell Type ◽

Tumor Mass ◽

Initial Cell ◽

Cloning Assay ◽

Human Tumor Cloning ◽

The Relationship

The in vitro human tumor cloning assay originally described by Salmon and Hamburger has been applied recently to the investigation of differential anti-tumor drug sensitivities over a broad range of human neoplasms. A major problem in the acceptance of this technique has been the question of the relationship between the cultured cells and the original patient tumor, i.e., whether the colonies that develop derive from the neoplasm or from some other cell type within the initial cell population. A study of the ultrastructural morphology of the cultured cells vs. patient tumor has therefore been undertaken to resolve this question. Direct correlation was assured by division of a common tumor mass at surgical resection, one biopsy being fixed for TEM studies, the second being rapidly transported to the laboratory for culture.

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Electron Probe Microanalysis of Frozen Hydrated Kidneys, Isolated Cells, and Cultured Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100054595 ◽

1982 ◽

Vol 40 ◽

pp. 402-403 ◽

Cited By ~ 1

Author(s):

Claude Lechene

Keyword(s):

Liquid Nitrogen ◽

Electron Probe Microanalysis ◽

Electron Probe ◽

Cultured Cells ◽

Liquid Nitrogen Temperature ◽

Elemental Content ◽

Isolated Cells ◽

Renal Tubular Cells ◽

Diamond Saw ◽

Saw Blade

Electron probe microanalysis of frozen hydrated kidneysThe goal of the method is to measure on the same preparation the chemical elemental content of the renal luminal tubular fluid and of the surrounding renal tubular cells. The following method has been developed. Rat kidneys are quenched in solid nitrogen. They are trimmed under liquid nitrogen and mounted in a copper holder using a conductive medium. Under liquid nitrogen, a flat surface is exposed by sawing with a diamond saw blade at constant speed and constant pressure using a custom-built cryosaw. Transfer into the electron probe column (Cameca, MBX) is made using a simple transfer device maintaining the sample under liquid nitrogen in an interlock chamber mounted on the electron probe column. After the liquid nitrogen is evaporated by creating a vacuum, the sample is pushed into the special stage of the instrument. The sample is maintained at close to liquid nitrogen temperature by circulation of liquid nitrogen in the special stage.

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