Confocal microscopy of cytoplasmic lipid droplets in a live cancer cell: number, polarity, diffusion and solvation dynamics

MedChemComm ◽  
2014 ◽  
Vol 5 (4) ◽  
pp. 536 ◽  
Author(s):  
Rajdeep Chowdhury ◽  
Batakrishna Jana ◽  
Abhijit Saha ◽  
Surajit Ghosh ◽  
Kankan Bhattacharyya
Keyword(s):  
Confocal Microscopy ◽  
Cancer Cell ◽  
Lipid Droplets ◽  
Cell Number ◽  
Solvation Dynamics ◽  
Cytoplasmic Lipid Droplets ◽  
Live Cancer Cell

126 CRYOPRESERVATION OF PORCINE EMBRYOS DERIVED FROM IVM OOCYTES

2007 ◽  
Vol 19 (1) ◽  
pp. 180
Author(s):  
N. Nakayama ◽  
K. Hiruma ◽  
M. Kurome ◽  
R. Tomii ◽  
S. Ueno ◽  
...  
Keyword(s):  
Zona Pellucida ◽  
Lipid Droplets ◽  
Cell Number ◽  
High Rate ◽  
Cell Stage ◽  
Cryoprotective Agents ◽  
Stage 4 ◽  
Morula Stage ◽  
Cytoplasmic Lipid Droplets

We have reported that a combination of delipation (removal of cytoplasmic lipid droplets from blastomeres) and vitrification by means of the minimum-volume cooling (MVC) method successfully cryopreserves porcine in vitro-matured/fertilized (IVM/IVF) embryos, and that normal piglets are produced from these embryos (Hiruma et al. 2006 Reprod. Fertil. Dev. 18, 157). We have also reported that IVM-derived embryos that undergo noninvasive delipation (i.e. micromanipulation is not required) and vitrification develop into blastocysts at a high rate (Esaki et al. 2004 Biol. Reprod. 71, 432–437). In this study, we examined whether fetuses can be produced from the IVM-derived embryos that have been delipated noninvasively and vitrified. Cumulus–oocyte complexes that had been collected from slaughterhouse ovaries were in vitro-matured in NCSU23 medium. The IVM oocytes were activated to produce parthenogenetic embryos. We used the embryos at the 4- to 8-cell (67 h after activation) and morula (98 h) stages in the following experiments. Embryos were treated with 4% trypsin (in PBS) at 38�C for 1 to 4 min to expand the zona pellucida. Next, the embryos were centrifuged (12 000g, 38�C, 23 min) in TL-HEPES-PVP containing 7.5 �g mL-1 cytochalasin B to polarize cytoplasmic lipid droplets within the perivitelline space. These embryos were cultured for 1 to 3 h and then vitrified. The post-thaw viability of the embryos was assessed based on their ability to develop into blastocysts and fetuses (21 to 23 days old). The embryos were vitrified using the MVC method with 15% ethylene glycol, 15% DMSO, and 0.5 M sucrose as cryoprotective agents. PZM-5 was used for culturing the embryos. In embryo transfer experiments, after thawing, the embryos were cultured for 36 or 72 h until they developed into morulae or 4- to 8-cell blastocysts, respectively; they were then treated with 0.5% pronase to remove the zona pellucida, and transferred to the uterine horns of estrus-synchronized recipients 6 days after onset of estrus. The proportion of vitrified embryos that developed into blastocysts and the mean cell number of the blastocysts were similar to those of non-vitrified control embryos, irrespective of the embryonic stage (4- to 8-cell stage: 42.1%, 22/51, 63.0 � 7.8 vs. 64.7%, 22/34, 74.2 � 7.1, respectively; morula stage: 77.6%, 38/49, 69.6 � 7.2 vs. 83.3%, 45/54, 66.2 � 5.9, respectively). Seventeen embryos that had been vitrified at the 4- to 8-cell stage gave rise to 3 fetuses after transfer into one recipient (17.6%). Fifty-three embryos that had been vitrified at the morula stage were transferred into 3 recipients. All recipients became pregnant and produced a total of 17 fetuses (32.1%). These results suggest that porcine IVM-derived embryos that have been cryopreserved by the combination of noninvasive delipation and vitrification by the MVC method are highly viable.

The lipid composition of isolated cytoplasmic lipid droplets from a human cancer cell line, BE(2)M17

2012 ◽  
Vol 8 (6) ◽  
pp. 1694 ◽  
Author(s):  
Xiaoyan Pan ◽  
Martin Wilson ◽  
Carmel McConville ◽  
Marie-Anne Brundler ◽  
Theodoros N. Arvanitis ◽  
...  
Keyword(s):  
Cell Line ◽  
Cancer Cell ◽  
Lipid Composition ◽  
Lipid Droplets ◽  
Human Cancer ◽  
Cancer Cell Line ◽  
Human Cancer Cell Line ◽  
Human Cancer Cell ◽  
Cytoplasmic Lipid Droplets

Vitrification of porcine early cleavage stage embryos and oocytes after removal of cytoplasmic lipid droplets

1996 ◽  
Vol 45 (1) ◽  
pp. 180 ◽  
Author(s):  
H. Nagashima ◽  
M. Kuwayama ◽  
C.G. Grupen ◽  
R.J. Ashman ◽  
M.B. Nottle
Keyword(s):  
Lipid Droplets ◽  
Cleavage Stage ◽  
Early Cleavage ◽  
Cytoplasmic Lipid Droplets

A proposal to identify live cancer cell by naked eye: Realization of biomedical application using 1D photonic structure

Optik ◽  
2019 ◽  
Vol 183 ◽  
pp. 818-821
Author(s):  
Iraj Sadegh Amiri ◽  
P. Yupapin ◽  
G. Palai ◽  
S.K. Tripathy
Keyword(s):  
Cancer Cell ◽  
Biomedical Application ◽  
Photonic Structure ◽  
Naked Eye ◽  
Live Cancer Cell

scholarly journals Recent Progress in Cryopreservation of Bovine Oocytes

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
In-Sul Hwang ◽  
Shinichi Hochi
Keyword(s):  
Lipid Droplets ◽  
Coiled Coil ◽  
High Sensitivity ◽  
Short Term ◽  
Rock Inhibitor ◽  
Developmental Potential ◽  
Domestic Species ◽  
Cytoplasmic Lipid Droplets ◽  
Recovery Culture ◽  
Bovine Oocytes

Principle of oocyte cryoinjury is first overviewed and then research history of cryopreservation using bovine oocytes is summarized for the last two decades with a few special references to recent progresses. Various types of cryodevices have been developed to accelerate the cooling rate and applied to the oocytes from large domestic species enriched with cytoplasmic lipid droplets. Two recent approaches include the qualitative improvement of IVM oocytes prior to the vitrification and the short-term recovery culture of vitrified-warmed oocytes prior to the subsequent IVF. Supplementation of L-carnitine to IVM medium of bovine oocytes has been reported to reduce the amount of cytoplasmic lipid droplets and improve the cryotolerance of the oocytes, but it is still controversial whether the positive effect of L-carnitine is reproducible. Incidence of multiple aster formation, a possible cause for low developmental potential of vitrified-warmed bovine oocytes, was inhibited by a short-term culture of the postwarm oocytes in the presence of Rho-associated coiled-coil kinase (ROCK) inhibitor. Use of an antioxidantα-tocopherol, instead of the ROCK inhibitor, also supported the revivability of the postwarm bovine oocytes. Further improvements of the vitrification procedure, combined with pre- and postvitrification chemical treatment, would overcome the high sensitivity of bovine oocytes to cryopreservation.

Studies of t6/t6 mouse embryos

Development ◽  
1975 ◽  
Vol 33 (3) ◽  
pp. 697-713
Author(s):  
Mary Nadijcka ◽  
Nina Hillman
Keyword(s):  
Lipid Droplets ◽  
Mouse Embryos ◽  
Wild Type ◽  
Electron Microscopic ◽  
Homozygous Mouse ◽  
Cytoplasmic Lipid Droplets

The phenocritical period of the t6/t6 genome extends from the late blastocyst substages through the elongated egg-cylinder stage, whereas the lethal period begins at the short egg-cylinder-stage and extends through the elongated egg-cylinder stage. Most of the homozygous mouse embryos die during the short egg-cylinder stage. The viable egg-cylinder-staged t6/t6 embryos can be distinguished from their phenotypically wild-type litter-mates at both the light and electron microscopic levels. The distinguishing characteristics of these embryos are aberrantly arranged entodermal cells, excessive cytoplasmic lipid and crystal-containing mitochondria. These same features are also characteristic of those mutant embryos which are developmentally arrested at both the short and elongated egg-cylinder stages. Over 50% of the t6/t6 embryos can be identified as early as the late blastocyst substages by the presence of large, electron-dense cytoplasmic lipid droplets.

scholarly journals Chimeric antigen receptors that trigger phagocytosis

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Meghan A Morrissey ◽  
Adam P Williamson ◽  
Adriana M Steinbach ◽  
Edward W Roberts ◽  
Nadja Kern ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Cancer Cell ◽  
Immune Cell ◽  
Human Cancer ◽  
Antibody Fragment ◽  
Cell Number ◽  
Antigen Receptors ◽  
Chimeric Antigen Receptors ◽  
Cell Therapies ◽  
Human Cancer Cells

Chimeric antigen receptors (CARs) are synthetic receptors that reprogram T cells to kill cancer. The success of CAR-T cell therapies highlights the promise of programmed immunity and suggests that applying CAR strategies to other immune cell lineages may be beneficial. Here, we engineered a family of Chimeric Antigen Receptors for Phagocytosis (CAR-Ps) that direct macrophages to engulf specific targets, including cancer cells. CAR-Ps consist of an extracellular antibody fragment, which can be modified to direct CAR-P activity towards specific antigens. By screening a panel of engulfment receptor intracellular domains, we found that the cytosolic domains from Megf10 and FcRɣ robustly triggered engulfment independently of their native extracellular domain. We show that CAR-Ps drive specific engulfment of antigen-coated synthetic particles and whole human cancer cells. Addition of a tandem PI3K recruitment domain increased cancer cell engulfment. Finally, we show that CAR-P expressing murine macrophages reduce cancer cell number in co-culture by over 40%.

47 EFFECT OF L-CARNITINE TREATMENT DURING OOCYTE MATURATION ON THE POST-THAW DEVELOPMENT OF PORCINE EMBRYOS VITRIFIED AT THE PRONUCLEAR STAGE

2016 ◽  
Vol 28 (2) ◽  
pp. 153 ◽  
Author(s):  
C. G. Grupen ◽  
T. Somfai ◽  
K. Kikuchi
Keyword(s):  
Lipid Droplets ◽  
In Vitro Maturation ◽  
Survival Rates ◽  
Cell Number ◽  
Blastocyst Formation ◽  
Porcine Embryo ◽  
Porcine Oocyte ◽  
Proportional Data ◽  
Bovine Oocytes

The extreme cryo-sensitivity of porcine oocytes and embryos is attributed to their endemically high content of cytoplasmic lipid droplets. In attempts to improve the cryo-tolerance of porcine embryos, various strategies have been used to reduce the amount of lipid droplets present in the cytoplasm before vitrification. Recently, the cryo-tolerance of bovine oocytes vitrified at the metaphase II stage was improved by supplementing in vitro maturation (IVM) medium with l-carnitine (LC), a stimulator of lipid metabolism (Chakitisakul et al. 2013 Theriogenology 79, 590–598). The objective of this study was to determine the effect of supplementing IVM medium with LC on the post-thaw development of porcine embryos vitrified at the pronuclear stage. Oocytes recovered from the ovaries of prepubertal gilts were matured in modified porcine oocyte medium supplemented with 0 (control) or 12 mM LC during the final 22 h of IVM. Following IVF, presumptive zygotes were cultured in porcine zygote medium-3. At the pronuclear stage, cohorts of embryos from each group were either vitrified using a solid surface vitrification procedure (Somfai et al. 2009 Biol. Reprod. 80, 42–49) or cultured for 7 d without being vitrified. Vitrified zygotes were subsequently warmed and cultured for 7 d. The rates of cleavage, blastocyst formation, and hatching were recorded, and all blastocysts were stained to determine the total cell numbers. Three replicates were performed. Proportional data were arcsine transformed and subjected to ANOVA, and cell number data were analysed by t-test. The post-thaw survival rates of the embryos that were vitrified did not differ between the groups (control: 95.7%; LC: 97.1%; P > 0.05). There were no significant effects of LC treatment or vitrification on the rates of cleavage, blastocyst formation, and hatching (Table 1). Vitrified embryos derived from LC-treated oocytes produced blastocysts with fewer cells than vitrified embryos derived from untreated oocytes (Table 1; P < 0.05). In contrast to previous findings in other species, the results indicate that supplementing IVM medium with LC did not enhance the post-thaw development of porcine embryos vitrified at the pronuclear stage. Table 1.Effect of l-carnitine (LC) treatment and vitrification on porcine embryo development C. Grupen was supported by an OECD Fellowship.

scholarly journals Multiplex coherent anti-Stokes Raman scattering microspectroscopy detection of lipid droplets in cancer cells expressing TrkB

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Tiffany Guerenne-Del Ben ◽  
Vincent Couderc ◽  
Ludovic Duponchel ◽  
Vincent Sol ◽  
Philippe Leproux ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Lipid Droplets ◽  
Live Cells ◽  
Label Free ◽  
Colorectal Cancer Cell Lines ◽  
Invasive Method ◽  
Dye Staining ◽  
Cytoplasmic Lipid Droplets ◽  
Anti Stokes ◽  
Cell Fixation

Abstract For many years, scientists have been looking for specific biomarkers associated with cancer cells for diagnosis purposes. These biomarkers mainly consist of proteins located at the cell surface (e.g. the TrkB receptor) whose activation is associated with specific metabolic modifications. Identification of these metabolic changes usually requires cell fixation and specific dye staining. MCARS microspectroscopy is a label-free, non-toxic, and minimally invasive method allowing to perform analyses of live cells and tissues. We used this method to follow the formation of lipid droplets in three colorectal cancer cell lines expressing TrkB. MCARS images of cells generated from signal integration of CH2 stretching modes allow to discriminate between lipid accumulation in the endoplasmic reticulum and the formation of cytoplasmic lipid droplets. We found that the number of the latter was related to the TrkB expression level. This result was confirmed thanks to the creation of a HEK cell line which over-expresses TrkB. We demonstrated that BDNF-induced TrkB activation leads to the formation of cytoplasmic lipid droplets, which can be abolished by K252a, an inhibitor of TrkB. So, MCARS microspectroscopy proved useful in characterizing cancer cells displaying an aberrant lipid metabolism.

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