scholarly journals Microprocessor-controlled vs. "dump-freezing" platelet and lymphocyte cryopreservation: A quantitative and qualitative comparative study

2006 ◽  
Vol 63 (3) ◽  
pp. 261-268 ◽  
Author(s):  
Bela Balint ◽  
Dusan Vucetic ◽  
Biljana Draskovic ◽  
Danilo Vojvodic ◽  
Goran Brajuskovic ◽  
...  
Keyword(s):  
Functional Recovery ◽  
Trypan Blue ◽  
Proliferative Response ◽  
Antigen Expression ◽  
Intergroup Difference ◽  
Cell Recovery ◽  
Trypan Blue Exclusion ◽  
Rate System ◽  
Platelet Recovery ◽  
Cell Percentage

Background/Aim. Thermodynamical and cryobiological parameters responsible for cell damages during cryopreservation (cryoinjuries) have not yet been completely explained. Thus, freezing procedures should be revised, exactly optimized to obtain an enhanced structural and functional recovery of frozen- thawed cells. The aim of this study was to compare microprocessor- controlled (controlled-rate) with the compensation of the released fusion heat and ?dump-freezing? (uncontrolled- rate) of the platelet and lymphocyte cryopreservation efficacy. Methods. Platelet quantitative recovery (post-thaw vs. unfrozen cell count), viability (using hypotonic shock response - HSR), morphological score (PMS), ultrastructural (electron microscopy) properties and expression of different surface antigens were investigated. In lymphocyte setting, cell recovery and viability (using trypan blue exclusion test) as well as functionality (by plant mitogens) were determined. Controlled- rate freezing and uncontrolled-rate cryopreservation were combined with 6% (platelets) and 10% (lymphocytes) dimethyl sulfoxide (DMSO). Results. Platelet recovery and functionality were superior in the controlled-rate system. The majority of surface antigen expression was reduced in both freezing groups vs. unfrozen cells, but GP140/CD62p was significantly higher in controlled-rate vs. uncontrolled-rate setting. Controlled- rate freezing resulted with better lymphocyte recovery and viability (trypan blue-negative cell percentage). In mitogen-induced lymphocyte proliferative response no significant intergroup difference (controlled-rate vs. uncontrolled-rate) were found. Conclusion. The data obtained in this study showned the dependence of cell response on the cryopreservation type. Controlled-rate freezing provided a superior platelet quantitative and functional recovery. Lymphocyte recovery and viability were better in the controlled-rate group, although only a minor intergroup difference for cell proliferative response was obtained.

Effects of medium and hypothermic temperatures on preservation of isolated porcine testis cells

2010 ◽  
Vol 22 (3) ◽  
pp. 523 ◽  
Author(s):  
Yanfei Yang ◽  
Ali Honaramooz
Keyword(s):  
Trypan Blue ◽  
Cell Isolation ◽  
Live Cell ◽  
Live Cells ◽  
Cell Recovery ◽  
Trypan Blue Exclusion ◽  
Culture Studies ◽  
Porcine Testis ◽  
Peritubular Cells

The effects of medium and hypothermic temperatures on testis cells were investigated to develop a strategy for their short-term preservation. Testes from 1-week-old piglets were enzymatically dissociated for cell isolation. In Experiment 1, testis cells were stored at either room (RT) or refrigeration (RG) temperature for 6 days in one of 13 different media. Live cell recovery was assayed daily using trypan blue exclusion. In Experiment 2, three media at RG were selected for immunocytochemical and in vitro culture studies. Live cell recovery was also assayed daily for 6 days using both trypan blue exclusion and a fluorochrome assay kit. For all media tested, significantly or numerically more live cells were maintained at RG than RT. On preservation Day 3 at RG (cell isolation day as Day 0), 20% FBS-Leibovitz resulted in the highest live cell recovery (89.5 ± 1.7%) and DPBS in the lowest (60.3 ± 1.9%). On Day 6 at RG, 20% FBS- Leibovitz also resulted in the best preservation efficiency with 80.9 ± 1.8% of Day 0 live cells recovered. There was no difference in live cell recovery detected by the two viability assays. After preservation, the proportion of gonocytes did not change, whereas that of Sertoli and peritubular cells increased and decreased, respectively. After 6 days of hypothermic preservation, testis cells showed similar culture potential to fresh cells. These results show that testis cells can be preserved for 6 days under hypothermic conditions with a live cell recovery of more than 80% and after-storage viability of 88%.

Polydopamine and dopamine interfere with tetrazolium-based cytotoxicity assays and produce exaggerated cytocompatibility inferences

2021 ◽  
Author(s):  
Anand Kumar Awasthi ◽  
Sakshi Gupta ◽  
Kavthekar Rupesh Namdev ◽  
Aditi Banerjee ◽  
Aasheesh Srivastava
Keyword(s):  
Cell Viability ◽  
Trypan Blue ◽  
Reliable Estimate ◽  
Trypan Blue Exclusion ◽  
Cytotoxicity Assays ◽  
Trypan Blue Exclusion Assay

Polydopamine (PDA) and dopamine (DA) can spontaneously reduce MTT reagent to formazan, resulting in incorrect cell-viability inferences. The non-redox Trypan Blue exclusion assay provides a more reliable estimate of cell viability with PDA and DA.

scholarly journals PHYTOCHEMICAL STUDY AND THE ANTIPROLIFERATIVE ACTIVITY OF INULA VULGARIS SPECIES GROWN IN LEBANON

2017 ◽  
Vol 9 (8) ◽  
pp. 75
Author(s):  
Assi M. ◽  
Usta J. ◽  
Mounimne Y. ◽  
Aboul Ela M. ◽  
El Lakany A.
Keyword(s):  
Ethyl Acetate ◽  
Antiproliferative Activity ◽  
Trypan Blue ◽  
Heart Diseases ◽  
Trypan Blue Exclusion ◽  
Potent Effect ◽  
Phytochemical Study ◽  
Chemical Structures ◽  
13C Nuclear Magnetic Resonance ◽  
Mtt Assays

Objective: Cancer represents the second leading cause of death after stroke and heart diseases. Plant extracts have long been used in traditional medicine for the prevention and treatment of many illnesses, including some types of cancer. The aim of this study was to evaluate the antiproliferative effects of ethyl acetate fractions of two Lebanese herbs: Inula viscosa (I. vis) and Inula vulgaris (I. vul).Methods: Plants were extracted with ethanol followed by ethyl acetate, then dried and tested on three cell lines including CaCO2, HepG2, and MCF7, to check for their viability and antiproliferative activity, using trypan blue exclusion and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Proton (1H) and carbon (13C) nuclear magnetic resonance spectrometry (NMR) were used to identify the compounds that have been isolated from both Inula species.Results: The current findings were consistent for both trypan blue and MTT assays. The results showed that the most potent effect for I. vul was HepG2 (IC50 20 µg/ml, 27 µg/ml), and for I. vis on MCF7 (9 µg/ml, 15 µg/ml) and CaCO2 (12 µg/ml, 22 µg/ml) in the two mentioned assays respectively. However, insignificant differences were observed among the studied plants for each of the evaluated cells indicating comparable potencies. Quercetin, quercetin glycoside, and epicatechin derivatives were isolated by fractionation on column chromatography and identified using NMR spectroscopy.Conclusion: The antiproliferative activities of the two plants could be related to their content that is significant for high levels of secondary metabolites. The identification of those compounds is necessary to establish a relationship between their chemical structures and their activities.

Digitonin-permeabilization of astrocytes in culture monitored by trypan blue exclusion and loss of S100B by ELISA

2000 ◽  
Vol 6 (1-2) ◽  
pp. 86-90 ◽  
Author(s):  
Francine Tramontina ◽  
Juliana Karl ◽  
Carmem Gottfried ◽  
Andreas Mendez ◽  
Daniela Gonçalves ◽  
...  
Keyword(s):  
Trypan Blue ◽  
Trypan Blue Exclusion ◽  
Digitonin Permeabilization

scholarly journals Fluorescence-activated sorting of rat hepatocytes based on their mixed function oxidase activities towards diethoxyfluorescein

1987 ◽  
Vol 247 (1) ◽  
pp. 23-28 ◽  
Author(s):  
I N H White ◽  
M L Green ◽  
R F Legg
Keyword(s):  
Dna Synthesis ◽  
Trypan Blue ◽  
Rat Hepatocytes ◽  
Fold Increase ◽  
Mixed Function Oxidase ◽  
Unscheduled Dna Synthesis ◽  
Rich Region ◽  
Trypan Blue Exclusion ◽  
Isolated Rat Hepatocytes ◽  
Viable Cells

The formation of ethoxyfluorescein and fluorescein from diethoxyfluorescein by isolated rat hepatocytes has been used as a basis for separating such cells dependent on their mixed function oxidase activities by fluorescence-activated flow cytometry. Five equal fractions defined by computer-generated regions were isolated. Non-viable cells with low fluorescence (region 1) represented 10-15% of the population, while the remainder with higher mixed function oxidase activities (regions 2-5), were greater than 95% viable by Trypan Blue exclusion. In region 1, 30% of the viable cells were binucleate, 67% diploid while in region 5, 13% were binucleate and 69% tetraploid. At 3 h after sorting, following attachment to glass coverslips, exposure of cells to methyl methanesulphonate, retrorsine or norethindrone resulted in unscheduled DNA synthesis which was 2-fold higher in the tetraploid-rich region 5, while aflatoxin B1, benzo[a]pyrene or 2-acetylaminofluorene caused a 5-fold increase in unscheduled DNA synthesis in these cells, relative to the diploid-rich hepatocytes in region 2.

In vitro photoradiation therapy of the rat 9L gliosarcoma

1986 ◽  
Vol 64 (5) ◽  
pp. 775-779 ◽  
Author(s):  
Douglas Hamilton ◽  
John D. S. McKean ◽  
John Tulip ◽  
Donald Boisvert ◽  
Judy Cummins
Keyword(s):  
Trypan Blue ◽  
Glioma Cells ◽  
Laser Source ◽  
Trypan Blue Exclusion ◽  
Content Type ◽  
9L Gliosarcoma ◽  
Trypan Blue Exclusion Test ◽  
9L Glioma ◽  
Fine Print

✓ The authors have investigated various factors involved in the photoradiation treatment of 9L glioma cells. The cells were grown in tissue culture and exposed to light from a laser source that allowed accurate quantitation of the light energy. Cell death was determined following treatment using the trypan blue exclusion test. It was shown that the treatment is very wavelength-dependent following the absorption spectrum of hematoporphyrin derivative (HPD). The absorption peaks in the lower part of the spectrum are more efficient than those of higher wavelengths. Photoradiation therapy is more effective the higher the concentration of HPD. Intensity of light is a very important factor in calculating the total dose of light necessary for this treatment.

scholarly journals Purification of erythroblastic nests

Blood ◽  
1981 ◽  
Vol 57 (5) ◽  
pp. 922-927
Author(s):  
AJ Macario ◽  
C Dugan ◽  
IL Perez-Lloret ◽  
E Conway de Macario
Keyword(s):  
Spleen Cell ◽  
Trypan Blue ◽  
Single Cells ◽  
Cell Suspensions ◽  
Differential Centrifugation ◽  
Minimum Essential Medium ◽  
Trypan Blue Exclusion ◽  
Large Numbers ◽  
Very High

We describe a method for preparing purified erythroblastic nests in large numbers (approximately 10(6)/run) in three steps: (1) induction of splenic erythropoiesis in mice, (2) preparative differential centrifugation for the removal of erythrocytes and single cells from spleen cell suspensions, and (3) sedimentation in an isokinetic gradient of Ficoll 400 in Joklik's modification of minimum essential medium. Viability of isolated EN is very high, as demonstrated by the trypan blue exclusion and in vitro erythrocyte formation methods.

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