Altered steroidogenic pattern of human granulosa-lutein cells in relation to cumulus cell culture morphology

1990 ◽  
Vol 36 (5) ◽  
pp. 457-464 ◽  
Author(s):  
Hela Gitay-Goren ◽  
Joseph M. Brandes ◽  
Shalom Bar-Ami
Keyword(s):  
Cell Culture ◽  
Cumulus Cell ◽  
Culture Morphology

10-gingerol induces oxidative stress through HTR1A in cumulus cells: in-vitro and in-silico studies

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Kiptiyah Kiptiyah ◽  
Widodo Widodo ◽  
Gatot Ciptadi ◽  
Aulanni’am Aulanni’Am ◽  
Mohammad A. Widodo ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Culture ◽  
Superoxide Dismutase ◽  
In Silico ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Sod Activity ◽  
In Silico Studies ◽  
Incubation Periods

AbstractBackgroundWe investigated whether 10-gingerol is able to induce oxidative stress in cumulus cells.MethodsFor the in-vitro research, we used a cumulus cell culture in M199, containing 10-gingerol in various concentrations (0, 12, 16, and 20 µM), and detected oxidative stress through superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentrations, with incubation periods of 24, 48, 72, and 96 h. The obtained results were confirmed by in-silico studies.ResultsThe in-vitro data revealed that SOD activity and MDA concentration increased with increasing incubation periods: SOD activity at 0 µM (1.39 ± 0.24i), 12 µM (16.42 ± 0.35ab), 16 µM (17.28 ± 0.55ab), 20 µM (17.81 ± 0.12a), with a contribution of 71.1%. MDA concentration at 0 µM (17.82 ± 1.39 l), 12 µM (72.99 ± 0.31c), 16 µM (79.77 ± 4.19b), 20 µM (85.07 ± 2.57a), with a contribution of 73.1%. Based on this, the in-silico data uncovered that 10˗gingerol induces oxidative stress in cumulus cells by inhibiting HTR1A functions and inactivating GSK3B and AKT˗1.Conclusions10-gingerol induces oxidative stress in cumulus cells through enhancing SOD activity and MDA concentration by inhibiting HTR1A functions and inactivating GSK3B and AKT˗1.

P–236 The evaluation of apoptosis and luteinization process in cumulus cell culture of IVF patients in terms of embryo development and pregnancy

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
G Dünda. Çiftlik ◽  
M Ergüven ◽  
T İrez
Keyword(s):  
Cell Culture ◽  
Embryo Development ◽  
Cell Apoptosis ◽  
Embryo Quality ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Clinical Pregnancy ◽  
Male Factor ◽  
Apoptosis Rate ◽  
Negative Effect

Abstract Study question Is there any relationship between apoptosis rate and progesterone levels in terms of the degree of embryo quality and clinical pregnancy success? Summary answer The cumulus cell apoptosis and progesterone levels measured at the hCG day were associated with embryo quality and clinical pregnancy success. What is known already The high rate of apoptosis at cumulus cells led to poor embryo development with the clinical pregnancy failure. Study design, size, duration It is prospective study carried out with 40 healthy women diagnosed as the male factor between September 2017 and April 2018. Participants/materials, setting, methods 40 healthy women aged between 27–38 and diagnosed as the male factor who were undergoing IVF participated the study. The cumulus cells were taken after denudation and cultured. The rate of the apoptosis were measured using with TUNEL method every 24 hours for 96 hours. In concominant with the apoptosis evaluation, the embryo progession and clinical pregnancy rate were also monitored. Progesterone levels taken at the hCG day were also measured. Main results and the role of chance In this study, it was shown that cumulus cell apoptosis was lower in the pregnant group compared to the non-pregnant group (p = 0.023), and progesterone value on the hCG day was higher in non-pregnant group (p = 0.021). From the data obtained at the end of the 4th day of culture showed that the apoptosis rate showed a higher tendency in non-pregnant women and this was statistically significant (p = 0.009). In our study, it was found that the ratio of apoptotic cells up to the 4th day showed a positive correlation with the progesterone levels on the hCG day (p = 0.001). Briefly, these results were obtainedfrom this current study that 1) The increase in the rate of apoptotic cells of cumulus cell culture has a negative effect on pregnancy rates, 2) The progesterone levels measured on the day of hCG has a significant negative effect on pregnancy, 3) The increase in progesterone levels acts as a messenger about the apoptosis rate of cumulus cells, 4) The luteinization is strictly associated with the apoptosis of cumulus cells. Limitations, reasons for caution: This was a single center study. Results need to be validated across different centers and high numbered different study population. Wider implications of the findings: The increase in progesterone value acts as a messenger about the rate of apoptosis of cumulus cells may be the prominent indicator to obtain high grade embryo with high clinical pregnancy success. Trial registration number 2017/5–8

scholarly journals The Predictive Value of the Pentraxin 3 Concentration in Cumulus Cell Culture Media for the Embryo Implantation

2021 ◽  
pp. 1-7
Author(s):  
Tulay Irez ◽  
Yavuz Sahin ◽  
Eduard Malik ◽  
Onur Guralp
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Culture Media ◽  
Embryo Implantation ◽  
Cumulus Cell ◽  
Culture Fluid ◽  
Cumulus Cells ◽  
Pentraxin 3 ◽  
Cell Culture Media ◽  
Morphological Criteria

OBJECTIVE: Many studies on the interrelation of cumulus cells and oocytes, and research on cumulus protein factors continue. This study aims to investigate the relationship of Pentraxin 3 level with embryo implantation in the cumulus culture fluid. STUDY DESIGN: A total of 31 women with idiopathic infertility who underwent intracytoplasmic sperm injection treatment were prospectively evaluated. Cell suspensions containing 5 million/mL cumulus cells were obtained post-hyase and incubated for 24 hours in a culture medium. A possible association between the culture media Pentraxin 3 concentrations and embryo implantation was analyzed. RESULTS: The cumulus cell culture media Pentraxin 3 concentrations were significantly higher in the pregnant group compared to the non-pregnant group (98.9 ng/mL vs 53.2 ng/mL, respectively, p=0.005). There was a significant positive correlation between the culture media Pentraxin 3 concentrations and embryo implantation (r=0.500, p=0.005). The culture media Pentraxin 3 concentration was a significant predictor for successful embryo implantation (AUC=0.845, p=0.006). A cut-off value of 64.25 ng/mL had an 86% sensitivity and 80% specificity to predict embryo implantation. There was no pregnancy under the cut-off value of 60 ng/mL, whereby seven women had good quality grade 1 oocytes according to the traditional morphological criteria. CONCLUSION: The cumulus cell culture medium Pentraxin 3 concentration was predictive for successful embryo implantation. Low Pentraxin 3 levels (<60 ng/mL were associated with failure of conception.

A T.E.M. study of mouse mammary epithelial cell secretory differentiation cultured on collagen and Matrigel

1991 ◽  
Vol 49 ◽  
pp. 188-189
Author(s):  
W.N. Bentham ◽  
V. Rocha
Keyword(s):  
Cell Culture ◽  
Mammary Epithelial ◽  
Secretory Process ◽  
Cell Culture System ◽  
Protein Maturation ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
Mouse Mammary Epithelial Cell ◽  
Secretory Differentiation

It has been an interest of our lab to develop a mammary epethelial cell culture system that faithfully duplicates the in vivo condition of the lactating gland. Since the introduction of collagen as a matrix on which cells are cultivated other E.C.M. type matrices have been made available and are used in many cell culture techniques. We have previously demonstrated that cells cultured on collagen and Matrigel do not differentiate as they do in vivo. It seems that these cultures often produce cells that show a disruption in the secretory process. The appearance of large ribosomal studded vesicles, that specifically label with antibody to casein, suggest an interruption of both protein maturation and secretion at the E.R. to golgi transition. In this report we have examined cultures on collagen and Matrigel at relative high and low seeding densities and compared them to cells from the in vivo condition.

Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses

1984 ◽  
Vol 42 ◽  
pp. 226-227 ◽  
Author(s):  
K. Pegg-Feige ◽  
F. W. Doane
Keyword(s):  
Electron Microscopy ◽  
Cell Culture ◽  
Immunoelectron Microscopy ◽  
Detection Method ◽  
Solid Phase ◽  
Protein A ◽  
Plant Viruses ◽  
Rapid Diagnosis ◽  
Virus Diagnosis ◽  
Antiviral Antibody

Immunoelectron microscopy (IEM) applied to rapid virus diagnosis offers a more sensitive detection method than direct electron microscopy (DEM), and can also be used to serotype viruses. One of several IEM techniques is that introduced by Derrick in 1972, in which antiviral antibody is attached to the support film of an EM specimen grid. Originally developed for plant viruses, it has recently been applied to several animal viruses, especially rotaviruses. We have investigated the use of this solid phase IEM technique (SPIEM) in detecting and identifying enteroviruses (in the form of crude cell culture isolates), and have compared it with a modified “SPIEM-SPA” method in which grids are coated with protein A from Staphylococcus aureus prior to exposure to antiserum.

Automated cell counting of astrocytes on patterned substrates containing aliphatic and charged properties

1995 ◽  
Vol 53 ◽  
pp. 974-975
Author(s):  
W. Shain ◽  
H. Ancin ◽  
H.C. Craighead ◽  
M. Isaacson ◽  
L. Kam ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Attachment ◽  
Culture Method ◽  
Cell Counting ◽  
Data Sets ◽  
Nuclear Staining ◽  
Double Positive ◽  
A Cell ◽  
Wafer Test ◽  
Cell Densities

Neural protheses have potential to restore nervous system functions lost by trauma or disease. Nanofabrication extends this approach to implants for stimulating and recording from single or small groups of neurons in the spinal cord and brain; however, tissue compatibility is a major limitation to their practical application. We are using a cell culture method for quantitatively measuring cell attachment to surfaces designed for nanofabricated neural prostheses.Silicon wafer test surfaces composed of 50-μm bars separated by aliphatic regions were fabricated using methods similar to a procedure described by Kleinfeld et al. Test surfaces contained either a single or double positive charge/residue. Cyanine dyes (diIC18(3)) stained the background and cell membranes (Fig 1); however, identification of individual cells at higher densities was difficult (Fig 2). Nuclear staining with acriflavine allowed discrimination of individual cells and permitted automated counting of nuclei using 3-D data sets from the confocal microscope (Fig 3). For cell attachment assays, LRM5 5 astroglial cells and astrocytes in primary cell culture were plated at increasing cell densities on test substrates, incubated for 24 hr, fixed, stained, mounted on coverslips, and imaged with a 10x objective.

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