Development of preovulatory follicles and oocytes during the oestrous cycle of mature and aged rats

1982 ◽  
Vol 100 (3) ◽  
pp. 434-443 ◽  
Author(s):  
J. J. Peluso ◽  
R. Hutz ◽  
C. England-Charlesworth
Keyword(s):  
Cell Viability ◽  
Granulosa Cells ◽  
Trypan Blue ◽  
Antral Follicle ◽  
Oestrous Cycle ◽  
Lower Percentage ◽  
Aged Rats ◽  
Trypan Blue Exclusion ◽  
Preovulatory Follicles ◽  
Lower Fertility

Abstract. The percentage of viable granulosa cells and the viability of the oocyte of each antral follicle on each day of the oestrous cycle of mature and aged rats was determined by trypan blue exclusion and fluorescein diacetate assays, respectively. In mature rats, the number of mid-sized follicles (350–550 μm in diameter) increased from oestrus through di-oestrus. The percentage of viable granulosa cells increased in only mid-sized follicles which contained a viable oocyte. On pro-oestrus, two types of preovulatory follicles (> 550 μm in diameter) were observed in the mature pro-oestrous rat: those with a high granulosa cell viability (> 60%) and a viable oocyte and those with a lower percentage of viable granulosa cells (< 50%) and a non-viable oocyte. This suggests that in mature rats only viable oocytes would be ovulated since presumably, only those preovulatory follicles with a high percentage of viable granulosa cells are able to ovulate. In aged rats, preovulatory-sized follicles were present on each day of the cycle. The percentage of viable granulosa cells within those preovulatory-sized follicles with nonviable oocytes increased on met-oestrus, decreased on di-oestrus and increased by pro-oestrus. This pattern resulted in the development of 3 to 4 preovulatory follicles per ovary with all preovulatory-sized follicles possessing a high percentage of viable granulosa cells regardless of the functional state of the oocyte. It is proposed that in aged rats, all the preovulatory follicles would ovulate but many of the ovulated oocytes would be defective. This mechanism would account for the lower fertility of these older animals.

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 2,3,7,8-Tetrachlorodibenzo-p-dioxin alters steroid secretion but does not affect cell viability and the incidence of apoptosis in porcine luteinised granulosa cells

2014 ◽  
Vol 62 (3) ◽  
pp. 408-421 ◽  
Author(s):  
Olga Jablonska ◽  
Joanna Piasecka-Srader ◽  
Anna Nynca ◽  
Agnieszka Kołomycka ◽  
Anna Robak ◽  
...  
Keyword(s):  
Cell Viability ◽  
Granulosa Cells ◽  
Steroid Hormone ◽  
Annexin V ◽  
Steroid Production ◽  
Steroid Secretion ◽  
Porcine Granulosa Cells ◽  
Affect Cell Viability ◽  
Preovulatory Follicles ◽  
Induced Apoptosis

The compound 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a by-product of human industrial activity, was found to affect ovarian steroidogenesis in animals, but the mechanism of its action is still unclear. The aims of the study were to examine the effect of TCDD on (1) progesterone (P4) and oestradiol (E2) production by granulosa cells isolated from medium (3–6 mm) and preovulatory (≥ 8 mm) porcine follicles, (2) the viability of the cells, and (3) the incidence of apoptosis. Porcine granulosa cells were cultured (48 h) with or without TCDD (100 pM, 100 nM). Steroid hormone concentrations in the medium were determined by radioimmunoassay. The viability of granulosa cells was tested spectrophotometrically (alamarBlue™ assay). Apoptosis was evaluated by flow cytometry using Annexin V and by TUNEL assay. The higher dose of TCDD (100 nM) significantly inhibited P4 and stimulated E2 production by luteinised granulosa cells isolated from medium follicles. The lower dose of TCDD (100 pM) significantly stimulated P4 and inhibited E2 secretion by the cells isolated from preovulatory follicles. None of the two TCDD doses affected cell viability or induced apoptosis in granulosa cells. In conclusion, TCDD directly affected steroid production by granulosa cells obtained from mature pigs, but the effect of TCDD was not due to its cytotoxicity.

scholarly journals Inhibitory Effects of Indirubin Derivatives on the Growth of HL-60 Leukemia Cells

2010 ◽  
Vol 5 (1) ◽  
pp. 1934578X1000500
Author(s):  
Nguyen Manh Cuong ◽  
Bui Huu Tai ◽  
Dang Hoang Hoan ◽  
Tran Thu Huong ◽  
Young Ho Kim ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cancer Cells ◽  
Trypan Blue ◽  
Promyelocytic Leukemia ◽  
Leukemia Cells ◽  
Cytotoxic Activities ◽  
Inhibitory Effects ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Method ◽  
Exclusion Method

Six indirubin derivatives have been synthesized and their inhibitory effects on the growth of HL-60 human promyelocytic leukemia cells investigated. Cell viability was determined using the trypan blue exclusion method. Indirubin-3′-oxime (I-1) inhibited the growth of HL-60 cells with a GI50 value of 36.6 μM, whereas I-0, I-2, I-3, I-4 and I-6 showed only weak cytotoxic activities against HL-60 cancer cells with GI50 values in the range of 97.3 to over 100 μM. These results indicate that indirubin derivatives might be useful candidate agents for exploring potential antileukemic drugs.

scholarly journals Trypan Blue Exclusion Assay Verifies in Vitro Cytotoxicity of New Cis-Platinum (II) Complex in Human Cells

2019 ◽  
Vol 16 (3) ◽  
pp. 0555
Author(s):  
Hussein Et al.
Keyword(s):  
Cell Viability ◽  
Trypan Blue ◽  
Membrane Integrity ◽  
Platinum Complex ◽  
In Vitro Cytotoxicity ◽  
Toxic Effects ◽  
Polymorphonuclear Cells ◽  
Trypan Blue Exclusion ◽  
High Concentrations

          Various assays are used to determine the toxic effects of drugs at cellular levels in vitro.  One of these methods is the dye exclusion assay, which measures membrane integrity in the presence of Trypan blue. Trypan blue the dye which was used in this study to investigate cytotoxic effect of a new Cis –dichloroplatinum (II) complex [(Qu)2PtCl2] on the viability of polymorphonuclear cells (PMNs). Three concentrations of platinum complex were prepared (70, 35and 17.5 µg/ ml) and the results revealed that the percentage of cell viability decreased as the platinum complex concentration increased in comparison with control. The platinum complex exhibited low cytotoxic effects towards healthy cells at the concentrations of 17.5 µg/ ml and 35 µg/ ml, in which the percentage of cell viability was (77.01 ± 6.3) and (72.3± 0.50)respectively, with no significant differences as compared with the control(90.66 ±0.577). The viability was significantly decreased (67.59 ± 3.16) when the cells were treated with the concentration of 70 µg/ ml in comparison with control. These results indicated that the percentage of living cells decreased when treated with high concentrations of [(Qu)2PtCl2], which causes cells death, while low concentrations of the compound show low toxicity. This data indicates that this compound, at these concentrations may be suitable for use as a cancer treatment because it has low toxic effects on the healthy cells.        

scholarly journals Variation in antral follicle development during the follicular phase of the oestrous cycle in red deer (Cervus elaphus) hinds

Reproduction ◽  
2001 ◽  
pp. 697-705 ◽  
Author(s):  
BJ McLeod ◽  
LM Meikle ◽  
MW Fisher ◽  
TR Manley ◽  
DA Heath ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Red Deer ◽  
Antral Follicle ◽  
Follicular Phase ◽  
Oestrous Cycle ◽  
Aromatase Activity ◽  
Follicle Development ◽  
Follicle Diameter ◽  
Follicle Size

The aim of this study was to quantify antral follicle populations in cyclic red deer hinds and to monitor follicle development leading to ovulation. Oestrus was synchronized with exogenous progesterone and ovaries were recovered approximately 0, 12, 24 or 36 h (follicular phase) or 10 days (luteal phase) after progesterone withdrawal (n = 5 per group). All follicles > or = 2 mm in diameter were dissected out, health status was assessed, follicular fluid oestradiol content was measured, granulosa cells were harvested and their capacity for oestradiol and cAMP production was determined. The time of oestrus and the preovulatory LH surge were monitored in five control hinds. Deer ovaries contained 26.6 +/- 3.45 (mean +/- SEM) follicles > or = 2 mm in diameter (range 4-81), with at least one large antral follicle (diameter: 8.3 +/- 0.38 mm) per hind. There was a strong correlation between follicle size and granulosa cell population (r(2) = 0.676). Approximately half (50.7%) of the follicles were classified as healthy, with the percentage classified as atretic decreasing with increasing follicle size. Neither the total number of antral follicles nor their size distribution differed significantly among groups. There were significantly more (P < 0.05) healthy follicles at 24 h after progesterone withdrawal than at 0 h, when large oestrogenic follicles had fewer granulosa cells, lower follicular fluid oestradiol concentrations and lower aromatase activity (P < 0.05) than did those from other groups. In summary, antral follicle development in red deer is similar to that in other monovulatory ruminants, and at least one large follicle is present at all stages of the oestrous cycle.

scholarly journals Induction of Necrosis and DNA Fragmentation during Hypothermic Preservation of Hepatocytes in UW, HTK, and Celsior Solutions

2003 ◽  
Vol 12 (1) ◽  
pp. 59-68 ◽  
Author(s):  
Salomon L. Abrahamse ◽  
Pieter Van Runnard Heimel ◽  
Robin J. Hartman ◽  
Rob A. F. M. Chamuleau ◽  
Thomas M. Van Gulik
Keyword(s):  
Cell Death ◽  
Cell Viability ◽  
Dna Fragmentation ◽  
Trypan Blue ◽  
Caspase 3 ◽  
Trypan Blue Exclusion ◽  
Hypothermic Preservation ◽  
Mtt Reduction ◽  
Ldh Release ◽  
Porcine Hepatocytes

Donor cells can be preserved in University of Wisconsin (UW), histidine-tryptophan-ketoglutarate (HTK), or Celsior solution. However, differences in efficacy and mode of action in preventing hypothermia-induced cell injury have not been unequivocally clarified. Therefore, we investigated and compared necrotic and apoptotic cell death of freshly isolated primary porcine hepatocytes after hypothermic preservation in UW, HTK, and Celsior solutions and subsequent normothermic culturing. Hepatocytes were isolated from porcine livers, divided in fractions, and hypothermically (4°C) stored in phosphate-buffered saline (PBS), UW, HTK, or Celsior solution. Cell necrosis and apoptosis were assessed after 24- and 48-h hypothermic storage and after 24-h normothermic culturing following the hypothermic preservation periods. Necrosis was assessed by trypan blue exclusion, lactate dehydrogenase (LDH) release, and mitochondrial 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) reduction. Apoptosis was assessed by the induction of histone-associated DNA fragments and cellular caspase-3 activity. Trypan blue exclusion, LDH release, and MTT reduction of hypothermically preserved hepatocytes showed a decrease in cell viability of more than 50% during the first 24 h of hypothermic preservation. Cell viability was further decreased after 48-h preservation. DNA fragmentation was slightly enhanced in hepatocytes after preservation in all solutions, but caspase-3 activity was not significantly increased in these cells. Normothermic culturing of hypothermically preserved cells further decreased cell viability as assessed by LDH release and MTT reduction. Normothermic culturing of hypothermically preserved hepatocytes induced DNA fragmentation, but caspase-3 activity was not enhanced in these cells. Trypan blue exclusion, LDH leakage, and MTT reduction demonstrated the highest cell viability after storage in Celsior, and DNA fragmentation was the lowest in cells that had been stored in PBS and UW solutions. None of the preservation solutions tested in this study was capable of adequately preventing cell death of isolated porcine hepatocytes after 24-h hypothermic preservation and subsequent 24-h normothermic culturing. Culturing of isolated and hypothermically preserved hepatocytes induces DNA fragmentation, but does not lead to caspase-3 activation. With respect to necrosis and DNA fragmentation of hypothermically preserved cells, UW and Celsior were superior to PBS and HTK solutions in this model of isolated porcine hepatocyte preservation.

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