Experimental results on mammalian cells growing in vitro in deuterated medium for neutron-scattering studies

1977 ◽  
Vol 25 (1) ◽  
pp. 87-94
Author(s):  
J. Murphy ◽  
C. Desaive ◽  
W. Giaretti ◽  
F. Kendall ◽  
C. Nicolini
Keyword(s):  
Mammalian Cells ◽  
Doubling Time ◽  
Deuterium Oxide ◽  
Initial Period ◽  
Lag Phase ◽  
Suspended Cell ◽  
Human Diploid Fibroblasts ◽  
Cell Doubling Time ◽  
Sv 40 Virus

SV-40 virus-transfromed human diploid fibroblasts (2RA) were grown in a monolayer on plastic Petri dishes in an aqueous medium deuterated to different concentrations of deuterium oxide: 10, 20, 30, up to 60%. The cells must be acclimatized to concentrations higher than 20% D2O by stepping them from a lower initial concentration during their exponential growth. The increase of cell doubling time with increasing deuterium concentrations seems to correlate, at least at 20% D20, with an initial period of suspended cell growth (lag-phase), and is qualitatively similar to that previously reported for Escherichia coli.

scholarly journals The Cultivation of Human Granulosa Cells

2008 ◽  
Vol 51 (3) ◽  
pp. 165-172 ◽  
Author(s):  
Lenka Brůčková ◽  
Tomáš Soukup ◽  
Jiří Moos ◽  
Martina Moosová ◽  
Jana Pavelková ◽  
...  
Keyword(s):  
Growth Factors ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Doubling Time ◽  
Ovarian Follicle ◽  
Vitro Fertilization ◽  
Thecal Cells ◽  
Cell Doubling Time

The major functions of granulosa cells (GCs) include the production of steroids, as well as a myriad of growth factors to interact with the oocyte during its development within the ovarian follicle. Also FSH stimulates GCs to convert androgens (coming from the thecal cells) to estradiol by aromatase. However, after ovulation the GCs produce progesterone that may maintain a potential pregnancy. Experiments with human GCs are mainly focused on the purification of GCs from ovarian follicular fluid followed by FACS analysis or short-term cultivation. The aim of our study was to cultivate GCs for a long period, to characterize their morphology and phenotype. Moreover, we have cultivated GCs under gonadotropin stimulation in order to simulate different pathological mechanisms during folliculogenesis (e.g. ovarian hyperstimulation syndrome). GCs were harvested from women undergoing in vitro fertilization. Complex oocyte-cumulus oophorus was dissociated by hyaluronidase. The best condition for transport of GCs was optimized as short transport in follicular fluid at 37 °C. GCs expansion medium consisted of DMEM/F12, 2 % FCS, ascorbic acid, dexamethasone, L-glutamine, gentamycine, penicillin, streptomycin and growth factors (EGF, bFGF). GCs transported in follicular fluid and cultivated in 2 % FCS containing DMEM/F12 medium supplemented with follicular fluid presented increased adhesion, proliferation, viability and decreased doubling time. Cell viability was 92 % and mean cell doubling time was 52 hrs. We have optimized transport and cultivation protocols for long-term cultivation of GCs.

scholarly journals Hyperglycemia and hyperinsulinemia stimulate cervical cancer cell proliferation in vitro and in vivo study

2019 ◽  
Author(s):  
Miriam Gutiérrez-Gutiérrez ◽  
Ishell Aline Figueroa-Martínez ◽  
Rafael Jurado ◽  
Norma Uribe ◽  
Patricia García-López ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cervical Cancer ◽  
Cell Proliferation ◽  
Cancer Cell ◽  
Doubling Time ◽  
Xenograft Model ◽  
Cervical Cancer Cell ◽  
Diabetic Patients ◽  
Cell Doubling Time

Abstract Background: Diabetes mellitus and malignant tumor are the second and third causes of women death in Mexico. Hyperglycemia, insulin and insulin-like growth factor 1 are the main risk factors involved in cancer development in patient with diabetes. The aim of this study was to evaluate the effect of hyperglycemia and hyperinsulinemia over cell proliferation and tumor growth in cervical cancer. Methods: Cell proliferation, apoptosis and cell cycle of cervical cancer cell lines (HeLa, SiHa and CaSki) in presence of hyperglycemia and/or insulin were evaluated. Xenograft model for cervical cancer was done in diabetic female nu/nu mice; biochemical parameters, body weight, tumoral volume and cell doubling time were evaluated. Results: Hyperglycemia and hyperinsulinemia significantly increase cell proliferation and decreases apoptosis with no change in cell cycle. Insulin treatment increase tumor volume and diminish cell doubling time, this group also developed hyperinsulinemia and in Langerhans pancreatic islet hypertrophy; whereas, hyperglycemic groups show the same effects but in lesser degree than the insulin treated group. Conclusion: Glucose and insulin stimulates both, proliferation and tumoral growth of cervical cancer, so this should be a possible explanation for the low survival of diabetic patients with cervical cancer in compare to non-diabetic patients with cervical cancer.

Cleavage rate of mouse embryos in vivo and in vitro

Development ◽  
1970 ◽  
Vol 24 (1) ◽  
pp. 203-207
Author(s):  
Patricia Bowman ◽  
Anne McLaren
Keyword(s):  
Doubling Time ◽  
Reproductive Tract ◽  
Cell Number ◽  
Blastocyst Stage ◽  
Mouse Embryos ◽  
Cleavage Rate ◽  
Cell Doubling Time ◽  
Constant Cell

Mouse embryos (Q strain) developing in vivo from the 2-cell to the blastocyst stage showed a constant cell doubling time of about 10 h. Embryos cultured in vitro over the same period showed a rate of cleavage which was initially almost as great as in the reproductive tract, but subsequently declined to give a doubling time of about 24 h. Addition of oestrogen to the culture medium increased the diameter of blastocysts but did not increase cell number.

scholarly journals Cytotoxicity, Antiproliferative and Apoptotic Effect of n-Hexane Fraction of Lime Parasite (Dendrophtoe pentandra)

Molekul ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. 1
Author(s):  
Subandrate Subandrate ◽  
Masayu Farah Diba ◽  
Salni Salni ◽  
Triwani Triwani ◽  
Sri Nita
Keyword(s):  
Flow Cytometry ◽  
Cytotoxic Activity ◽  
Doubling Time ◽  
T47d Cell ◽  
Mtt Method ◽  
Folk Remedy ◽  
Proliferation And Apoptosis ◽  
Cell Doubling Time ◽  
Apoptotic Effect

Breast cancer is one of the biggest causes of death in women in the world. Lime parasite (Dendrophtoe pentandra (L.) Miq.), a folk remedy used by Indonesian people, is believed to be efficacious as anticancer drug. This research aims to know the activity of n-hexane fractions of lime parasite in inhibiting the proliferation and apoptosis of T47D cells in vitro. Cytotoxic test with MTT method assay from n-hexane fractions used a multilevel concentration. Antiproliferative test was carried out by the method of MTT assay and cell doubling time was calculated at the time of duplication. Apoptotic test was done with concentration of 1 IC50and ½ IC50which was analyzed by flow cytometry. The results reveals that fractions of lime parasite have cytotoxic activity with concentration of IC50is included in moderatecytotoxic level. The result of the doubling time of the optimum fraction of n-hexane is in 31 hours with the concentration of ¼IC50. Results for the flow cytometry shows the fraction of n-hexane does not induce apoptosis in cells of T47D.  Those results show that the active fraction of lime parasite has cytotoxic activity which is able to inhibit proliferation, but does not induce apoptosis of T47D cell.

scholarly journals Postnatal Changes of Neural Stem Cells in the Mammalian Auditory Cortex

2021 ◽  
Vol 22 (4) ◽  
pp. 1550
Author(s):  
Zhengqing Hu ◽  
Li Tao ◽  
Meng Deng
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Neural Stem Cells ◽  
Auditory Cortex ◽  
Postnatal Development ◽  
Doubling Time ◽  
Future Research ◽  
Cell Doubling Time

Our previous study reported neural stem cells (NSCs) in the auditory cortex (AC) of postnatal day 3 (P3) mice in vitro. It is unclear whether AC-NSCs exist in vivo. This study aims to determine the presence and changes of AC-NSCs during postnatal development and maturation both in vitro and in vivo. P3, postnatal day 14 (P14), 2-month-old (2M), and 4-month-old (4M) mouse brain tissues were fixed and cryosectioned for NSC marker immunostaining. In vitro, P3, P14, and 2M AC tissues were dissected and cultured in suspension to study NSCs. NSC proliferation was examined by EdU incorporation and cell doubling time assays in vitro. The results show that Nestin and Sox2 double expressing NSCs were observed in the AC area from P3 to 4M in vivo, in which the number of NSCs remarkably reduced with age. In vitro, the neurosphere forming capability, cell proliferation, and percentage of Nestin and Sox2 double expressing NSCs significantly diminished with age. These results suggest that AC-NSCs exist in the mouse AC area both in vitro and in vivo, and the percentage of AC-NSCs decreases during postnatal development and maturation. The results may provide important cues for the future research of the central auditory system.

Ultrastructural Changes in Three Different Mammalian Cells Following Ultraviolet Laser Irradiation

1972 ◽  
Vol 30 ◽  
pp. 350-351
Author(s):  
K. Shankar Narayan ◽  
Kailash C. Gupta ◽  
Tohru Okigaki
Keyword(s):  
Ultraviolet Light ◽  
Mammalian Cells ◽  
Biological Effects ◽  
Survival Rates ◽  
Chinese Hamster ◽  
Cell Types ◽  
Differential Sensitivity ◽  
Ultrastructural Changes ◽  
Ultraviolet Laser

The biological effects of short-wave ultraviolet light has generally been described in terms of changes in cell growth or survival rates and production of chromosomal aberrations. Ultrastructural changes following exposure of cells to ultraviolet light, particularly at 265 nm, have not been reported.We have developed a means of irradiating populations of cells grown in vitro to a monochromatic ultraviolet laser beam at a wavelength of 265 nm based on the method of Johnson. The cell types studies were: i) WI-38, a human diploid fibroblast; ii) CMP, a human adenocarcinoma cell line; and iii) Don C-II, a Chinese hamster fibroblast cell strain. The cells were exposed either in situ or in suspension to the ultraviolet laser (UVL) beam. Irradiated cell populations were studied either "immediately" or following growth for 1-8 days after irradiation.Differential sensitivity, as measured by survival rates were observed in the three cell types studied. Pattern of ultrastructural changes were also different in the three cell types.

Vacuoles Induced in Mammalian Cells by Helicobacter Cytotoxin are Acidic Organelles

1990 ◽  
Vol 48 (3) ◽  
pp. 168-169
Author(s):  
M. H. Chestnut ◽  
C. E. Catrenich
Keyword(s):  
Acridine Orange ◽  
Mammalian Cells ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Ulcer Disease ◽  
Gastroduodenal Disease ◽  
H Pylori

Helicobacter pylori is a non-invasive, Gram-negative spiral bacterium first identified in 1983, and subsequently implicated in the pathogenesis of gastroduodenal disease including gastritis and peptic ulcer disease. Cytotoxic activity, manifested by intracytoplasmic vacuolation of mammalian cells in vitro, was identified in 55% of H. pylori strains examined. The vacuoles increase in number and size during extended incubation, resulting in vacuolar and cellular degeneration after 24 h to 48 h. Vacuolation of gastric epithelial cells is also observed in vivo during infection by H. pylori. A high molecular weight, heat labile protein is believed to be responsible for vacuolation and to significantly contribute to the development of gastroduodenal disease in humans. The mechanism by which the cytotoxin exerts its effect is unknown, as is the intracellular origin of the vacuolar membrane and contents. Acridine orange is a membrane-permeant weak base that initially accumulates in low-pH compartments. We have used acridine orange accumulation in conjunction with confocal laser scanning microscopy of toxin-treated cells to begin probing the nature and origin of these vacuoles.

Cytological changes induced by platinum-thymine in sarcoma-180 cells: Electron microscopy study

1990 ◽  
Vol 48 (3) ◽  
pp. 290-291
Author(s):  
Gustav Ofosu
Keyword(s):  
Mammalian Cells ◽  
Antitumor Agent ◽  
Microscopy Study ◽  
Electron Microscopy Study ◽  
Limiting Factor ◽  
Sarcoma 180 ◽  
Electron Microscopic ◽  
Cytological Changes

Platinum-thymine has been found to be a potent antitumor agent, which is quite soluble in water, and lack nephrotoxicity as the dose-limiting factor. The drug has been shown to interact with DNA and inhibits DNA, RNA and protein synthesis in mammalian cells in vitro. This investigation was undertaken to elucidate the cytotoxic effects of piatinum-thymine on sarcoma-180 cells in vitro ultrastructurally, Sarcoma-180 tumor bearing mice were treated with intraperitoneal injection of platinum-thymine 40mg/kg. A concentration of 60μg/ml dose of platinum-thymine was used in in vitro experiments. Treatments were at varying time intervals of 3, 7 and 21 days for in vivo experiments, and 30, 60 and 120 min., 6, 12, and 24th in vitro. Controls were not treated with platinum-thymine.Electron microscopic analyses of the treated cells in vivo and in vitro showed drastic cytotoxic effect.

Programmable in vitro Co-Encapsidation of Guest Proteins Inside Protein Nanocages

2018 ◽  
Author(s):  
Noor H. Dashti ◽  
Rufika S. Abidin ◽  
Frank Sainsbury
Keyword(s):  
Self Assembly ◽  
Mammalian Cells ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Super Resolution ◽  
Cell Engineering ◽  
Particle Analysis ◽  
Protein Cages

Bioinspired self-sorting and self-assembling systems using engineered versions of natural protein cages have been developed for biocatalysis and therapeutic delivery. The packaging and intracellular delivery of guest proteins is of particular interest for both <i>in vitro</i> and <i>in vivo</i> cell engineering. However, there is a lack of platforms in bionanotechnology that combine programmable guest protein encapsidation with efficient intracellular uptake. We report a minimal peptide anchor for <i>in vivo</i> self-sorting of cargo-linked capsomeres of the Murine polyomavirus (MPyV) major coat protein that enables controlled encapsidation of guest proteins by <i>in vitro</i> self-assembly. Using Förster resonance energy transfer (FRET) we demonstrate the flexibility in this system to support co-encapsidation of multiple proteins. Complementing these ensemble measurements with single particle analysis by super-resolution microscopy shows that the stochastic nature of co-encapsidation is an overriding principle. This has implications for the design and deployment of both native and engineered self-sorting encapsulation systems and for the assembly of infectious virions. Taking advantage of the encoded affinity for sialic acids ubiquitously displayed on the surface of mammalian cells, we demonstrate the ability of self-assembled MPyV virus-like particles to mediate efficient delivery of guest proteins to the cytosol of primary human cells. This platform for programmable co-encapsidation and efficient cytosolic delivery of complementary biomolecules therefore has enormous potential in cell engineering.

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