Grafted polymer brush coatings for growth of cow granulosa cells and oocyte-cumulus cell complexes

2020 ◽  
Vol 15 (3) ◽  
pp. 031006
Author(s):  
Yurij Stetsyshyn ◽  
Joanna Raczkowska ◽  
Khrystyna Harhay ◽  
Kamil Awsiuk ◽  
Yana Shymborska ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Polymer Brush ◽  
Cumulus Cell ◽  
Cell Complexes ◽  
Grafted Polymer

Mitochondrial cell-free DNA secreted from porcine granulosa cells

Zygote ◽  
2019 ◽  
Vol 27 (05) ◽  
pp. 272-278 ◽  
Author(s):  
Kazuki Kansaku ◽  
Yasuhisa Munakata ◽  
Koumei Shirasuna ◽  
Takehito Kuwayama ◽  
Hisataka Iwata
Keyword(s):  
Granulosa Cells ◽  
Culture Medium ◽  
Cumulus Cell ◽  
Oocyte Quality ◽  
Cell Free Dna ◽  
Cell Complexes ◽  
Free Dna ◽  
Porcine Granulosa Cells ◽  
Maturation Medium ◽  
Potential Biomarker

SummarySeveral studies have proposed that cell-free DNA (cfDNA) is a potential biomarker present in follicular fluid (FF) for oocyte quality. Recently we reported that mitochondria-derived cfDNA (mt-cfDNA) closely reflects the amount of cfDNA in FFs. The present study investigated the mechanism regulating mt-cfDNA secretion from porcine granulosa cells. Oocytes and cumulus cell complexes or granulosa cells (GCs) were cultured in maturation medium for 24 or 48 h respectively. Then, nuclear-derived cell-free DNA (n-cfDNA) or mt-cfDNA contents in the spent medium were examined using real-time polymerase chain reaction. When 10 μM of MG132, a proteasome inhibitor, was added to the culture medium, cellular viability of both COCs and GCs decreased and n-cfDNA significantly increased in the culture medium, whereas mt-cfDNA significantly decreased. Supplementation of the culture medium with GW4869, an inhibitor of intracellular vesicle formation, significantly decreased the mt-cfDNA, whereas no effect was observed on n-cfDNA in the medium of both COCs and GCs. Furthermore, the addition of bafilomycin, an inhibitor of autophagy to the culture medium significantly increased mt-cfDNA in the culture medium. After filtration (0.22 μm) and centrifugation (23,000 g), the mt-cfDNA content of the medium decreased significantly. In conclusion, the proteasomal mitochondrial quality control system is upstream of mt-cfDNA secretion and autophagy plays a role in cellular digestion of mitochondrial DNA in the cytoplasm. It is further suggested that dsDNA is enclosed in certain vesicles or associated with small molecules and secreted into the medium.

Light-Responsive Polymer Surfaces via Postpolymerization Modification of Grafted Polymer-Brush Structures

Langmuir ◽  
2014 ◽  
Vol 30 (49) ◽  
pp. 14971-14981 ◽  
Author(s):  
Matthias Dübner ◽  
Nicholas D. Spencer ◽  
Celestino Padeste
Keyword(s):  
Polymer Brush ◽  
Polymer Surfaces ◽  
Grafted Polymer ◽  
Responsive Polymer

037. IMMUNE-LIKE MECHANISMS ASSOCIATED WITH OVULATION

2009 ◽  
Vol 21 (9) ◽  
pp. 10
Author(s):  
J. Richards
Keyword(s):  
Granulosa Cells ◽  
Immune Cell ◽  
Expression Patterns ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Mature Oocyte ◽  
Innate Immune ◽  
Innate Immune Cell ◽  
Oocyte Meiosis ◽  
Mitogen Activated Protein

Ovulation is the unique biological process by which a mature oocyte and surrounding somatic cells, the cumulus cell-oocyte complex (COC), are released from the surface of the ovary into the oviduct for transport and fertilization. Ovulation is similar to an inflammatory response: the follicles become hyperemic, produce prostaglandins and synthesize a hyaluronan-rich extracellular matrix. However, this view of ovulation may be too restrictive and need to be broadened to encompass the innate immune cell surveillance response system. This hypothesis is being proposed because ovarian granulosa cells and cumulus cells express and respond to innate immune cell related surveillance proteins (Toll-like receptors 2 and 4) and cytokines such as interleukin 6 (IL6) during ovulation. In addition, recent studies indicate that the ovulation process that is set in motion by the surge of luteinizing hormone (LH) is mediated, in large part, by the EGF-like factors (Amphiregulin, epiregulin and betacellulin) and their critical activation of RAS, most probably KRAS that is expressed at high levels in granulosa cells, and the mitogen activated protein kinases, MAP3/1 (ERK1/2). Mice in which granulosa cells are depleted of ERK1/2 fail to ovulate, oocyte meiosis does not resume, COC expansion is impaired and luteinization is blocked. Thus the global molecular reprogramming of granulosa cell gene expression patterns is completely derailed. Supported, in part by NIH-HD-16229, -16272 and -07495 (SCCPIR).

scholarly journals Oil Deposition on Polymer Brush-Coated NF Membranes

Membranes ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 168 ◽  
Author(s):  
Anh Vu ◽  
Naama Segev Mark ◽  
Guy Z. Ramon ◽  
Xianghong Qian ◽  
Arijit Sengupta ◽  
...  
Keyword(s):  
Membrane Fouling ◽  
Membrane Surface ◽  
Polymer Brush ◽  
Operating Mode ◽  
Feed Solution ◽  
Polymer Chains ◽  
Membrane Properties ◽  
Solution Temperature ◽  
Oil Droplets ◽  
Grafted Polymer

Membrane-based processes are attractive for treating oily wastewaters. However, membrane fouling due to the deposition of oil droplets on the membrane surface compromises performance. Here, real-time observation of the deposition of oil droplets by direct confocal microscopy was conducted. Experiments were conducted in dead-end and crossflow modes. Base NF 270 nanofiltration membranes as well as membranes modified by grafting poly(N-isopropylacrylamide) chains from the membrane surface using atom transfer radical polymerization were investigated. By using feed streams containing low and high NaCl concentrations, the grafted polymer chains could be induced to switch conformation from a hydrated to a dehydrated state, as the lower critical solution temperature for the grafted polymer chains moved above and below the room temperature, respectively. For the modified membrane, it was shown that switching conformation of the grafted polymer chains led to the partial release of adsorbed oil. The results also indicate that, unlike particles such as polystyrene beads, adsorption of oil droplets can lead to coalescence of the adsorbed oil droplets on the membrane surface. The results provide further evidence of the importance of membrane properties, feed solution characteristics, and operating mode and conditions on membrane fouling.

Alterations in the cell cycle of mouse cumulus granulosa cells during expansion and mucification in vivo and in vitro

1996 ◽  
Vol 8 (6) ◽  
pp. 935 ◽  
Author(s):  
AW Schuetz ◽  
DG Whittingham ◽  
R Snowden
Keyword(s):  
Cell Cycle ◽  
Dna Synthesis ◽  
Granulosa Cells ◽  
Dna Content ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Ovarian Follicles ◽  
S Phase

The cell cycle characteristics of mouse cumulus granulosa cells were determined before, during and following their expansion and mucification in vivo and in vitro. Cumulus-oocyte complexes (COC) were recovered from ovarian follicles or oviducts of prepubertal mice previously injected with pregnant mare serum gonadotrophin (PMSG) or a mixture of PMSG and human chorionic gonadotrophin (PMSG+hCG) to synchronize follicle differentiation and ovulation. Cell cycle parameters were determined by monitoring DNA content of cumulus cell nuclei, collected under rigorously controlled conditions, by flow cytometry. The proportion of cumulus cells in three cell cycle-related populations (G0/G1; S; G2/M) was calculated before and after exposure to various experimental conditions in vivo or in vitro. About 30% of cumulus cells recovered from undifferentiated (compact) COC isolated 43-45 h after PMSG injections were in S phase and 63% were in G0/G1 (2C DNA content). Less than 10% of the cells were in the G2/M population. Cell cycle profiles of cumulus cells recovered from mucified COC (oviducal) after PMSG+hCG-induced ovulation varied markedly from those collected before hCG injection and were characterized by the relative absence of S-phase cells and an increased proportion of cells in G0/G1. Cell cycle profiles of cumulus cells collected from mucified COC recovered from mouse ovarian follicles before ovulation (9-10 h after hCG) were also characterized by loss of S-phase cells and an increased G0/G1 population. Results suggest that changes in cell cycle parameters in vivo are primarily mediated in response to physiological changes that occur in the intrafollicular environment initiated by the ovulatory stimulus. A similar lack of S-phase cells was observed in mucified cumulus cells collected 24 h after exposure in vitro of compact COC to dibutyryl cyclic adenosine monophosphate (DBcAMP), follicle-stimulating hormone or epidermal growth factor (EGF). Additionally, the proportion of cumulus cells in G2/M was enhanced in COC exposed to DBcAMP, suggesting that cell division was inhibited under these conditions. Thus, both the G1-->S-phase and G2-->M-phase transitions in the cell cycle appear to be amenable to physiological regulation. Time course studies revealed dose-dependent changes in morphology occurred within 6 h of exposure in vitro of COC to EGF or DBcAMP. Results suggest that the disappearance of the S-phase population is a consequence of a decline in the number of cells beginning DNA synthesis and exit of cells from the S phase following completion of DNA synthesis. Furthermore, loss of proliferative activity in cumulus cells appears to be closely associated with COC expansion and mucification, whether induced under physiological conditions in vivo or in response to a range of hormonal stimuli in vitro. The observations indicate that several signal-transducing pathways mediate changes in cell cycle parameters during cumulus cell differentiation.

scholarly journals The Nuclear Receptor Cofactor Receptor-Interacting Protein 140 Is a Positive Regulator of Amphiregulin Expression and Cumulus Cell-Oocyte Complex Expansion in the Mouse Ovary

Endocrinology ◽  
2010 ◽  
Vol 151 (6) ◽  
pp. 2923-2932 ◽  
Author(s):  
Jaya Nautiyal ◽  
Jennifer H. Steel ◽  
Meritxell M. Rosell ◽  
Evanthia Nikolopoulou ◽  
Kevin Lee ◽  
...  
Keyword(s):  
Nuclear Receptor ◽  
Granulosa Cells ◽  
Cumulus Cell ◽  
Proximal Promoter ◽  
Dependent Manner ◽  
Camp Response Element ◽  
Response Element ◽  
Cumulus Expansion ◽  
Interacting Protein

The nuclear receptor cofactor receptor-interacting protein 140 (RIP140) is essential for cumulus cell-oocyte complex (COC) expansion, follicular rupture, and oocyte release during ovulation. The expression of many genes necessary for COC expansion is impaired in the absence of RIP140, but the studies herein document that their expression can be restored and COC expansion rescued by treatment with the epidermal growth factor (EGF)-like factor amphiregulin (AREG) both in vitro and in vivo. We demonstrate by several approaches that RIP140 is required for the expression of the EGF-like factors in granulosa cells, but the dependence of genes involved in cumulus expansion, including Ptgs2 Has2, Tnfaip6, and Ptx3, is indirect because they are induced by AREG. Treatment of granulosa cells with forskolin to mimic the effects of LH increases AREG promoter activity in a RIP140-dependent manner that 1) requires an intact cAMP response element in the proximal promoter region of the Areg gene and 2) involves its actions as a coactivator for cAMP response element-binding protein/c-Jun transcription factors. Although human chorionic gonadotropin and AREG coadministration is sufficient to restore ovulation fully in RIP140 heterozygous mice in vivo, both follicular rupture and ovulation remain impaired in the RIP140 null mice. Thus, we conclude that although the level of RIP140 expression in the ovary is a crucial factor required for the transient expression of EGF-like factors necessary for cumulus expansion, it also plays a role in other signaling pathways that induce follicular rupture.

Effects of polydispersity in the end-grafted polymer brush

1989 ◽  
Vol 22 (2) ◽  
pp. 853-861 ◽  
Author(s):  
S. T. Milner ◽  
T. A. Witten ◽  
M. E. Cates
Keyword(s):  
Polymer Brush ◽  
Grafted Polymer
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