Effect of estradiol during culture of bovine oocyte–granulosa cell complexes on the mitochondrial DNA copies of oocytes and telomere length of granulosa cells

Zygote ◽  
2012 ◽  
Vol 22 (4) ◽  
pp. 431-439 ◽  
Author(s):  
M. Endo ◽  
K. Kimura ◽  
T. Kuwayama ◽  
Y. Monji ◽  
H. Iwata
Keyword(s):  
Mitochondrial Dna ◽  
Telomere Length ◽  
Granulosa Cells ◽  
Culture Medium ◽  
Mt Dna ◽  
Cell Complexes ◽  
Antral Follicles ◽  
Mitochondrial Dna Copy Number ◽  
Significant Difference

SummaryDuring the development of oocytes from early antral follicles (EAFs) to antral follicles (AFs), the mitochondrial DNA copy number (Mt DNA number) increases, and granulosa cells markedly proliferate. This study examined the effect of supplementation of culture medium with estradiol-17β (E2) on the in vitro growth of oocytes, and increases in the Mt DNA number, and telomere length during the in vitro culture of oocytes derived from EAFs (0.4–0.7 mm in diameter). The E2 supplementation improved antrum formation and the ratio of oocytes reaching the metaphase II (MII) stage, and there was a significant difference in these values between addition E2 concentrations of 10 μg/ml and 0.1 μg/ml. When the oocytes were cultured in the medium containing 10 μg/ml E2, the Mt DNA number determined by real-time polymerase chain reaction (PCR) significantly increased, and the ratio of the Mt DNA number at the end of culture to the Mt DNA number at the beginning of the culture was greatly different among cows, and could be predicted by the degree of the difference between the Mt DNA number of oocytes derived from EAFs and that of oocytes derived from AFs (3–6 mm in diameter). When oocytes were cultured for 16 days in a medium containing 10 μg/ml E2 or 0.1 μg/ml E2, the Mt DNA number of oocytes grown in vitro did not differ, but the telomere length of the granulosa cells was significantly greater in the 10 μg/ml E2 group than in the 0.1 μg/ml group. In conclusion, E2 supplementation in culture medium improved the growth of oocytes derived from EAFs, and a high E2 concentration increased the telomere length of the granulosa cells.

Addition of granulosa cell mass to the culture medium of oocytes derived from early antral follicles increases oocyte growth, ATP content, and acetylation of H4K12

Zygote ◽  
2016 ◽  
Vol 24 (6) ◽  
pp. 848-856 ◽  
Author(s):  
Miyako Sugiyama ◽  
Mei Sumiya ◽  
Koumei Shirasuna ◽  
Takehito Kuwayama ◽  
Hisataka Iwata
Keyword(s):  
Granulosa Cell ◽  
Granulosa Cells ◽  
Culture Medium ◽  
Cell Mass ◽  
Fertilization Rate ◽  
Atp Content ◽  
Oocyte Growth ◽  
Acetylation Level ◽  
Antral Follicles

SummaryThe main aim of the present study was to examine the hypothesis that an increase in the number of granulosa cells surrounding developing bovine oocytes results in both high ATP levels and an increase in the acetylation level of H4K12 in oocytes grown in vitro. Oocyte–granulosa cell complexes (OGCs) were collected from early antral follicles (EAFs, 0.4–0.7 mm in diameter), and individually cultured on 96-well plates with or without additional granulosa cell mass that had been prepared from other OGCs. After 16 days of culture, we examined: (i) the rate of antrum formation of the OGCs; (ii) the diameter, maturation, and fertilization rate of the oocytes; and (iii) the ATP content and acetylation level of H4K12 in the oocytes grown in vitro. Granulosa cell mass added to the culture medium contributed to the development of OGCs with a higher rate of antrum formation and oocyte growth. Furthermore, the addition of granulosa cells increased the ATP content and acetylation level of H4K12 in oocytes grown in vitro compared with those developed without addition of granulosa cells. In addition, there was a positive correlation between the ATP content in oocytes grown in vitro and the number of granulosa cells in the corresponding OGCs. The results suggest that granulosa cells play a role not only in the development of OGCs and the growth of oocytes, but also in the determination of ATP content and the acetylation of H4K12 in the oocytes developed in vitro.

163 IN VITRO CULTURE OF PORCINE OOCYTE-GRANULOSA CELL COMPLEXES

2011 ◽  
Vol 23 (1) ◽  
pp. 184
Author(s):  
Y. F. Diao ◽  
R. X. Han ◽  
H. R. Kim ◽  
C. S. Park ◽  
D. I. Jin
Keyword(s):  
Survival Rate ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Control Group ◽  
Oocyte Diameter ◽  
Cell Complexes ◽  
Treatment Groups ◽  
Significant Difference ◽  
Porcine Oocyte

The objective of this study was to investigate the effects of porcine follicular fluid (PFF) and insulin-like growth factor-1 (IGF-1) on the growth of porcine oocyte-granulosa cell complexes (OGC) in vitro, in an effort to improve meiotic and developmental competence in vitro. Porcine OGC were manually dissected from early antral follicles of diameter 400 to 700 μm, and intact oocytes with undamaged granulosa cells were selected for culture. Between 3 and 5 OGC were combined in 50-uL droplets and cultured for 12 days in M199 medium supplemented with PVP, oestradiol, FSH, transferrin, L-ascorbic acid, and insulin. The OGC were cultured at 38.5°C in a humidified atmosphere of 5% CO2 in air for 12 days, and the oocytes were matured for 44 h. Oocyte diameter, exclusive of the zona pellucida, was measured on day 0 and day 12 of culture. Control group was cultured in the absence of PFF or IGF-1. The experiment was divided into 2 parts. In part 1, treatment groups were cultured with 2.5, 5.0, or 7.5% (all v/v) PFF. Control OGC grew as spheres that were formed by granulosa cells. In treatment groups, the granulosa cells spread and grew on the bottom of dishes. When oocyte diameter was measured after 12 days of culture, no significant difference among groups was observed (104.07, 103.96, and 104.27 μm at the 3 PFF concentrations used; control: 104.03 μm). Similarly, the survival rate of oocytes did not differ significantly among groups. However, survival rate fell somewhat in the group treated with PFF (control: 65%; tests: 58.87, 58.33, and 50.83% at the 3 PFF concentrations used). The maturation rates of oocytes in the control was significantly higher than those of the treatment groups [25.83% (control) v. 14, 11.67, and 3.67% (the 3 treatment groups); P < 0.05]. Thus, the first conclusion is that supplementation of culture medium with PFF did not enhance the development of porcine OGC in vitro. In part 2 of the experiment, treatment groups were cultured with 10, 50, or 100 ng mL–1 IGF-1. The percentages of OGC showing antrum formation were 80, 80, 100, and 100% in groups treated with 0, 10, 50, or 100 ng mL–1 IGF-1, respectively. Average oocyte diameter was 94.16 to 94.58 μm just after OGC collection. However, the average diameter of oocytes cultured for 12 days with 50 or 100 ng mL–1 IGF-1 was significantly higher than that of the control or 10 ng mL–1 groups [108.88 and 108.31 μm (50 and 100 ng mL–1 IGF-1) v. 105.98 μm and 106.67 μm (0 and 10 ng mL–1 IGF-1); P < 0.05]. The maturation rate of oocytes grown with 10 and 50 ng mL–1 IGF-1 was higher than those of the other 2 groups [30 and 40.6% (10 and 50 ng mL–1 IGF-1, respectively) v. 25 and 26% (0 and 100 ng mL–1 IGF-1, respectively); P < 0.05]. Thus, the second conclusion is that 50 ng mL–1 IGF-1 improves the growth and maturation of porcine oocyte-granulosa cell complexes in vitro.

In vitro culture of oocytes with surrounding cumulus complexes and granulosa cells (COCGs) from bovine early antral follicles

2003 ◽  
Vol 77 (1) ◽  
pp. 141-147
Author(s):  
S. Saha ◽  
M. Shimizu ◽  
M. Geshi ◽  
Y. Izaike
Keyword(s):  
Growth Factor ◽  
Granulosa Cells ◽  
Culture Medium ◽  
Follicle Stimulating Hormone ◽  
Germinal Vesicle ◽  
Antral Follicles ◽  
Ovarian Cortex ◽  
Epidermal Growth ◽  
Germinal Vesicle Break Down

AbstractCumulus-oocyte complexes with surrounding granulosa cells (COCGs) in early antral follicles (0·5 to 0·7 mm in diameter) were surgically collected from sections of bovine ovarian cortex under a dissection microscope and subsequently cultured in vitro using follicle stimulating hormone (FSH), epidermal growth factor (EGF), insulin-transferrin-selenium (ITS) and hypoxanthine, singly or in combination, to obtain fully grown matured oocytes. Oocytes cultured in the presence of FSH + hypoxanthine increased (P < 0·05) in diameter from 93 µm on the day of commencement of culture to 106·37±0·34 µm on day 5. Oocytes cultured in the presence of FSH, hypoxanthine or hypoxanthine + ITS + FSH increased (P < 0·05) to mean diameters of 105·40 (s.e. 0·47) µm, 105·50 (s.e. 0·39) µm and 105·35 (s.e. 0·55) µm, respectively. By day 11 of culture, oocyte diameters 110·50 (s.e. 0·35) µm, 110·13 (s.e. 0·39) µm, 109·49 (s.e. 0·46) µm, 109·53 (s.e. 0·58) µm and 109·16 (s.e. 0·43) µm were recorded for treatments FSH + hypoxanthine, hypoxanthine + ITS + FSH, FSH, hypoxanthine and FSH + EGF + hypoxanthine + ITS, respectively. The proportions with COCGs which formed an antrum while cultured in vitro; were categorized as morphologically normal following recovery from the gel; matured in vitro; showed germinal vesicle break down and reached metaphase II were highest (P < 0·05) for the FSH + hypoxanthine treatment (49/60 (81·7%), 48/60 (80·0%), 47/60 (78·3%), 45/60 (75·0%) and 15/60 (25·0%), respectively, followed by hypoxanthine + ITS + FSH (47/60 (78·3%), 44/60 (73·3%), 41/60 (68·3%), 41/60 (68·3%) and 12/60 (20%), respectively), FSH (43/60 (71·7%), 42/60 (70%), 40/60 (66·7%), 39/60 (65·0%) and 9/60 (15%), respectively) and hypoxanthine (41/60 (68·3%), 38/60 (63·3%), 36/60 (60%), 35/60 (58·3%) and 8/60 (13·3%), respectively). In experiment II, the in vitro fertilization and cleavage rates of COCGs were highest (P < 0·05) for FSH + hypoxanthine treatment (17/60; 28·3%) followed by hypoxanthine + ITS + FSH (13/60; 21·6%), FSH (12/60; 20%) and hypoxanthine (11/60; 18·3%) treatments. The results of this study show that COCGs from early antral follicles can be isolated, cultured and grown in vitro. Furthermore, supplements like FSH and hypoxanthine can be used singly or in combination(s) in culture medium to enhance the growth of COCGs.

scholarly journals Promotion of follicular antrum formation by pig oocytes in vitro

Zygote ◽  
1998 ◽  
Vol 6 (1) ◽  
pp. 47-54 ◽  
Author(s):  
Xiangju Shen ◽  
Takashi Miyano ◽  
Seishiro Kato
Keyword(s):  
Granulosa Cells ◽  
Collagen Gels ◽  
Similar Extent ◽  
Intimate Contact ◽  
Cell Complexes ◽  
Antral Follicles ◽  
Pig Oocyte ◽  
Stimulation Of

SummaryPig oocyte–cumulus–granulosa cell complexes (OCG complexes) from pig early antral follicles reorganise an antrum under the stimulation of FSH. The purpose of this study was to examine the role of the oocytes in antrum formation. In the first experiment, oocyte–cumulus complexes were removed from pig OCG complexes, and the antrum formation of parietal granulosa cells themselves (PGs) was examined. Antrum formation by sham-operated OCG complexes (OC/G complexes), in which the connections between the oocyte–cumulus complexes and the parietal granulosa cells had been disrupted, was also examined. The complexes were cultured for 8 days in collagen gels in the presence of 10ng/ml FSH. Antra were formed in about 60% of the intact OCG complexes and the sham-operated OCG complexes, while only 20% of the PGs formed antra. In the second experiment, oocyte–cumulus complexes in the OCG complexes were replaced by denuded oocytes (O/G complexes) or Sephadex G-25 beads (B/G complexes) similar in diameter to the oocytes, and the two types of complexes were cultured under the same conditions. The O/G complexes formed antra to a similar extent as the OC/G complexes, whereas the B/G complexes scarcely formed any antra. The histological sections showed that the granulosa cells in the OC/G and O/G complexes were in intimate contact with each other and retained a shape similar to those in the ovarian follicles, while the granulosa cells in the PGs and B/G complexes became quite irregular in shape. These results suggest that pig oocytes promote contact between the granulosa cells to induce antrum formation in a physiological manner.

Should fertile women quit drinking alcohol to produce better quality oocytes?

Zygote ◽  
2020 ◽  
pp. 1-3
Author(s):  
Burcu Ozbakir ◽  
Pinar Tulay
Keyword(s):  
Alcohol Consumption ◽  
Young Women ◽  
Antral Follicle ◽  
Antral Follicle Count ◽  
Control Group ◽  
Social Drinkers ◽  
Antral Follicles ◽  
Healthy Women ◽  
Significant Difference

Summary Alcohol consumption has long been shown to affect both fetal health and pregnancy. In this study, antral follicle count, maturation level of oocytes including morphological assessment and number of metaphase I (MI), metaphase II (MII) and germinal vesicle (GV) stage oocytes obtained from young women (age < 30 years old) with or without alcohol consumption were investigated. In total, 20 healthy women who were social drinkers and 36 healthy women who do not consume alcohol were involved in this study. Women in both study and control groups were undergoing controlled ovarian stimulation. The antral follicle count and the number and quality of the oocytes retrieved were evaluated and recorded. In total, 635 antral follicles, 1098 follicles and 1014 oocytes with 820 MII, 72 MI and 78 GV stage oocytes were collected from the social drinkers. In the control group, 628 antral follicles, 1136 follicles and 1085 oocytes with 838 MII, 93 MI and 102 GV stage oocytes were evaluated. The results of this study showed that the antral follicle count was very similar in both groups. The number of oocytes and MII stage oocytes was slightly higher in the control group, although it was not a significant difference. This study showed that although the consumption of alcohol may have adverse effects post-implantation, it may not have a solid effect during oogenesis in young women. The results of this study are especially important in clinical settings as some women who are social drinkers undergo in vitro fertilization treatments.

Loss of the Association between Telomere Length and Mitochondrial DNA Copy Number Contribute to Colorectal Carcinogenesis

2017 ◽  
Vol 24 (2) ◽  
pp. 323-328 ◽  
Author(s):  
Hyunsu Lee ◽  
Ji-Hyoung Cho ◽  
Won-Jin Park ◽  
Soo-Jung Jung ◽  
In-Jang Choi ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Telomere Length ◽  
Copy Number ◽  
Colorectal Carcinogenesis ◽  
Dna Copy Number ◽  
Mitochondrial Dna Copy Number

scholarly journals Telomere length is correlated with mitochondrial DNA copy number in intestinal, but not diffuse, gastric cancer

2017 ◽  
Vol 14 (1) ◽  
pp. 925-929 ◽  
Author(s):  
Soo-Jung Jung ◽  
Ji-Hyoung Cho ◽  
Won-Jin Park ◽  
Yu-Ran Heo ◽  
Jae-Ho Lee
Keyword(s):  
Gastric Cancer ◽  
Mitochondrial Dna ◽  
Telomere Length ◽  
Copy Number ◽  
Diffuse Gastric Cancer ◽  
Dna Copy Number ◽  
Mitochondrial Dna Copy Number

scholarly journals PERIPHERAL BLOOD MITOCHONDRIAL DNA ALTERATION DURING SYSTEMIC LUPUS NEPHRITIS

2021 ◽  
Vol 10 (19) ◽  
pp. 416-421
Author(s):  
Ruchi Upadhyay ◽  
Ratika Srivastava
Keyword(s):  
Mitochondrial Dna ◽  
Lupus Nephritis ◽  
Lupus Erythematosus ◽  
Peripheral Blood ◽  
Normal Subjects ◽  
Mitochondrial Genes ◽  
Systemic Lupus ◽  
Mt Dna ◽  
Mitochondrial Dna Copy Number ◽  
Genes Encoding

The investigation of mitochondrial DNA (Mt-DNA) alterations could impart light on the involvement of mitochondria in the pathophysiology of Systemic Lupus Erythematosus. The purpose of this study is to examine the peripheral blood mitochondrial DNA copy number variation in Lupus Nephritis patients and also to find out it’s correlation with amount of protein present in urine. The significant correlation could aid in the inspection of mitochondrial involvement, particularly in Lupus Nephritis. Two mitochondrial genes encoding MT-CYT and MT-TL1 were measured quantitatively by qRT-PCR in whole blood of 17 SLE patients and 15 healthy subjects with similar gender (female: male ratio) and age group. The amount of mitochondrial genes MT-CYT and MT-TL1 was 1.69 and 1.26 fold higher respectively in patients. The significantly higher amount of protein detected in lupus nephritis patients (129.4±116.4 mg/dl) in comparison to normal subjects (25.3 ±10.7 mg/dl). No significant correlation was established between Mt-DNA quantity and proteinuria. Alteration in mitochondrial genes reflects the possibilities of altered mitophagy or mitochondrial biosynthesis during SLE. These findings are required to be further validated by studying mitophagy and biogenesis during SLE in details.

Sign in / Sign up

Export Citation Format

Share Document