maturation rate
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2022 ◽  
Vol 34 (2) ◽  
pp. 305
Author(s):  
M. D. Sebopela ◽  
M. L. Mphaphathi ◽  
S. M. Sithole ◽  
T. L. Nedambale

Author(s):  
Arya T. S. ◽  
Amritha Aravind ◽  
Abhilash R. S. ◽  
Jayakumar C. ◽  
Babitha V.

The present study was conducted to assess the developmental competence of goat oocytes selected using Brilliant cresyl blue (BCB) staining. Goat ovaries were collected from the slaughtered animals with unknown reproductive history. The oocytes retrieved by aspiration technique were selected based on morphology and subjected to BCB staining. Brilliant cresyl blue staining is based on the activity of glucose-6-phospahte dehydrogenase (G6PDH) enzyme synthesised by the oocytes. The cytoplasm remains blue in oocytes that have finished the growth phase (BCB+) while the growing oocytes remain colourless (BCB-). The stained and unstained oocytes were subjected to in vitro maturation separately to assess cumulus cell expansion index and polar body extrusion. A total of 206 culture grade oocytes were subjected to study, out of which, 76.75 ± 2.38 per cent of oocytes showed positive to BCB staining and 23.21 ± 2.38 per cent were negatively stained. Significantly higher maturation rate was observed in BCB+(92.89 ± 2.37%) oocytes than BCB-(29.72 ± 2.46%). The present study concluded that BCB staining can be used for selecting goat oocytes with good cytoplasmic maturation for further in vitro embryo production


Author(s):  
Amaneh Moradi ◽  
Fatemeh Ghasemian ◽  
Farhad Mashayekhi

Background: The dialogue between oocytes and their surrounding cells plays a major role in the progress of oocyte meiosis and their developmental potential. Objective: This study aimed to evaluate the effect of co-culture of normal granulosa-cumulus cells (GCCs) with oocytes from polycystic ovarian syndrome (PCOS) mice. Materials and Methods: Normal GCCs were collected from 10 virgin adult Naval Medical Research Institute female mice (30-35 gr, 7-8 wk old), and were cultured in an alpha-minimum essential medium supplemented with 5% fetal calf serum for 24-48 hr (1×106 cells/well). Then, germinal-vesicle oocytes from PCOS mice were cultured in the presence of cultured normal GCCs (experimental group) and without GCCs (control group). The maturation rate and quality of the PCOS oocytes were examined by evaluating TFAM and Cx43 gene expression (real-time PCR) and the connection among PCOS oocytes and normal GCCs after 24 hr of culture. Results: The co-culture of normal GCCs and PCOS oocytes in the experimental group led to the formation of a complex called a PCOS oocyte-normal GCCs complex. The maturation rate of these complexes was significantly increased compared to that of the control group (p ≤ 0.001). A significant difference was also found in the expression of Cx43 (p ≤ 0.001) and TFAM (p < 0.05) genes in the experimental group compared with the control group. The connection between PCOS oocytes and normal GCCs was observed in the scanning electron microscope images. Conclusion: Co-culture with normal GCCs improves the capacity of PCOS oocytes to enter meiosis, which may result in the promotion of assisted reproduction techniques. Key words: PCOS, Co-culture, Granulosa-cumulus cells, IVM, Cx43.


2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Qiaoqi Chen ◽  
Liang Zhang ◽  
Lin Li ◽  
Mixiao Tan ◽  
Weiwei Liu ◽  
...  

Abstract Background Mono-therapeutic modality has limitations in combating metastatic lesions with complications. Although emerging immunotherapy exhibits preliminary success, solid tumors are usually immunosuppressive, leading to ineffective antitumor immune responses and immunotherapeutic resistance. The rational combination of several therapeutic modalities may potentially become a new therapeutic strategy to effectively combat cancer. Results Poly lactic-co-glycolic acid (PLGA, 50 mg) nanospheres were constructed with photothermal transduction agents (PTAs)-Prussian blue (PB, 2.98 mg) encapsulated in the core and chemotherapeutic docetaxel (DTX, 4.18 mg)/ immune adjuvant-imiquimod (R837, 1.57 mg) loaded in the shell. Tumor cell membranes were further coated outside PLGA nanospheres (designated “M@P-PDR”), which acted as “Nano-targeted cells” to actively accumulate in tumor sites, and were guided/monitored by photoacoustic (PA)/ magnetic resonance (MR) imaging. Upon laser irradiation, photothermal effects were triggered. Combined with DTX, PTT induced in situ tumor eradication. Assisted by the immune adjuvant R837, the maturation rate of DCs increased by 4.34-fold compared with that of the control. In addition, DTX polarized M2-phenotype tumor-associated macrophages (TAMs) to M1-phenotype, relieving the immunosuppressive TME. The proportion of M2-TAMs decreased from 68.57% to 32.80%, and the proportion of M1-TAMs increased from 37.02% to 70.81%. Integrating the above processes, the infiltration of cytotoxic T lymphocytes (CTLs) increased from 17.33% (control) to 35.5%. Primary tumors and metastasis were significantly inhibited when treated with “Nano-targeted cells”-based cocktail therapy. Conclusion “Nano-targeted cells”-based therapeutic cocktail therapy is a promising approach to promote tumor regression and counter metastasis/recurrence. Graphical Abstract


2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Qiyu Yang ◽  
Lixia Zhu ◽  
Meng Wang ◽  
Bo Huang ◽  
Zhou Li ◽  
...  

Abstract Background To improve the developmental competence of in vitro cultured oocytes, extensive literature focused on maturation rate improvement with different additives in culture medium, while studies investigating the maturation dynamics of oocytes during in vitro maturation (IVM) and the influencing factors on oocyte viability are scarce. Methods The study involved a retrospective observation by time-lapse monitoring of the IVM process of 157 donated GV oocytes from 59 infertile couples receiving ICSI in 2019, in Tongji Hospital, Wuhan, China. The GV oocytes derived from controlled ovarian hyperstimulation (COH) cycles underwent rescue IVM (R-IVM), and the maturation dynamics, including GVBD time (GV-MI), time from GVBD to maturation (MI-MII), maturation time (GV-MII), and MII arrest duration (MII-ICSI), were recorded by time-lapse monitoring. The matured oocytes were inseminated at different MII arrest points and subsequent embryo developments were assessed. The effects of baseline clinical characteristics, oocyte diameters, and maturation dynamics on the developmental competence of the oocytes were also analyzed. Results Totally, 157 GV oocytes were collected. GVBD happened in 111 oocytes, with a median GV-MI duration of 3.7 h. The median MI-MII duration was 15.6 h and the median GV-MII duration was 19.5 h. The maturation rate reached 56.7% at 24 h and 66.9% at 48 h, and the clinical factors, including patient age, FSH level, AMH level, ovarian stimulation protocol, and serum estradiol and progesterone levels on hCG trigger day, showed no effects on the 24-h maturation rate. The normal fertilization rate of oocytes resuming meiosis within 8 h and matured within 24 h was significantly higher than that of oocytes resuming meiosis after 8 h and matured after 24 h. Furthermore, among those oocytes matured within 24 h, the high-quality embryo formation rate of oocytes resuming meiosis within 4.5 h and matured within 19 h was significantly higher. All stated time was measured from the start point of IVM. Additionally, for oocytes from patients with serum progesterone levels less than 1 ng/ml on hCG trigger day, the high-quality embryo formation rate was significantly increased. Conclusion R-IVM technology could increase the available embryos for patients in routine COH cycles, but excessive culture beyond 24 h is not recommended. GV-MI duration of the oocyte, recorded by time-lapse system, and serum progesterone levels of patients on hCG trigger day can significantly affect the developmental potential of the IVM oocytes.


2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Anton Ivanov Kolarov ◽  
Irina Valcheva Chakarova ◽  
Valentina Prodanova Hadzhinesheva ◽  
Venera Pantaleeva Nikolova ◽  
Stefka Metodieva Delimitreva ◽  
...  

Known as a degenerative joint disorder of advanced age affecting predominantly females, osteoarthritis can develop in younger and actively working people because of activities involving loading and injuries of joints. Collagenase-induced osteoarthritis (CIOA) in a mouse model allowed us to investigate for the first time its effects on key cytoskeletal structures (meiotic spindles and actin distribution) of ovulated mouse oocytes. Their meiotic spindles, actin caps, and chromatin were analyzed by immunofluorescence. A total of 193 oocytes from mice with CIOA and 209 from control animals were obtained, almost all in metaphase I (M I) or metaphase II (MII). The maturation rate was lower in CIOA (26.42% M II) than in controls (55.50% M II). CIOA oocytes had significantly larger spindles (average 37 μm versus 25 μm in controls, p < 0.001 ), with a proportion of large spindles more than 64% in CIOA versus up to 15% in controls ( p < 0.001 ). Meiotic spindles were wider in 68.35% M I and 54.90% M II of CIOA oocytes (mean 18.04 μm M I and 17.34 μm M II versus controls: 11.64 μm M I and 12.64 μm M II), and their poles were approximately two times broader (mean 6.9 μm) in CIOA than in controls (3.6 μm). CIOA oocytes often contained disoriented microtubules. Actin cap was visible in over 91% of controls and less than 20% of CIOA oocytes. Many CIOA oocytes without an actin cap had a nonpolarized thick peripheral actin ring (61.87% of M I and 52.94% of M II). Chromosome alignment was normal in more than 82% in both groups. In conclusion, CIOA affects the cytoskeleton of ovulated mouse oocytes—meiotic spindles are longer and wider, their poles are broader and with disorganized fibers, and the actin cap is replaced by a broad nonpolarized ring. Nevertheless, meiotic spindles were successfully formed in CIOA oocytes and, even when abnormal, allowed correct alignment of chromosomes.


2021 ◽  
Author(s):  
Yen-Ju Sung ◽  
Liang-Hsuan Chen ◽  
Tzu-Hsuan Chin ◽  
Shang-Yu Huang ◽  
Hsing-Tse Yu ◽  
...  

Abstract Background Evidently, when undergoing GnRH-antagonist protocols, dual trigger has proven to produce not just better quality and quantity of oocytes but also pregnancy outcome. However, not much comparative studies have been published when PPOS protocol is used for ovarian stimulation. Can the same positive outcomes be expected after the patients have been exposed to the high level of progesterone required for PPOS protocols? Methods In this retrospective cohort study, patients undergoing PPOS protocols were separated into three groups based on the method employed for triggering final follicular maturation, which included: (a) human chorionic gonadotropin (hCG); (b) Gonadotropin-releasing hormone-agonist (GnRH-agonist); or (c)dual trigger (GnRH-agonist + hCG). Either in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) was utilized for fertilization. Assessment comprised of their dynamic hormone profiles, embryonic analysis, and clinical outcomes. Results Of the 344 recruited patients, those fulfilling the Bologna criteria as poor ovarian responders and showing Estradiol (E2)<1000 pg/ml on the day of triggering had higher oocyte maturation rate (82% vs 58%, p<0.05) when triggered with dual trigger (GnRH-agonist + hCG) than hCG alone. For the patients with E2> 6500 pg/ml on the day of triggering, none of the three triggering methods demonstrated a significant advantage regarding the number of oocytes, percentage of matured oocytes, and rate of oocytes at fertilization or cleavage stages. Conclusions Implementing dual trigger for stimulating final follicular maturation in patients undergoing PPOS protocols is debatable. For poor ovarian response (POR) patients, dual trigger appeared to yield higher percentage of matured oocytes. In contrast, for hyper-responders, methods of triggering oocyte maturation did not affect the percentage of matured oocytes or the qualities of the embryos. For this group of patients, therefore, the agent used should be one that would reduce the risks of ovarian hyper-stimulation syndrome (OHSS).


2021 ◽  
Author(s):  
SHUANG TANG ◽  
Nannan Yang ◽  
Mingxi Yu ◽  
Shuo Wang ◽  
Xiangdong Hu ◽  
...  

Mitochondria transfer can rescue oocyte aging-related infertility. However, heterologous techniques are suspended due to heteroplasmy. Regarding autologous approaches, the donor source and manipulating procedures require further optimization. Here we propose a strategy using umbilical cord mesenchymal stem cells (UC-MSCs) as mitochondria donor cells and employing intercellular mitochondria transport as the transfer method. We cryopreserved UC-MSCs of the female pup. When the female aged, its UC-MSCs were induced into granulosa cells (iGCs). The zona-weakened GV oocytes were aggregated with autologous iGCs into iGC-oocyte complexes. After cultivation in GDF9-containing media, mitochondria migrated from iGCs into the GV oocyte via transzonal filopodia. The maturation rate, quality, and developmental potential of these oocytes were substantially increased. Furthermore, the birth rate after embryo transfer has been improved. This approach utilized noninvasive procedures to collect mitochondria donor cells and optimized mitochondria transfer manipulations, so may represent a promising advance towards the improvement of aging-related infertility.


Author(s):  
Demissie Tilaye ◽  
Yilma Tefera ◽  
Degefa Tamrat ◽  
Wirtu Gemechu ◽  
Lemma Alemayehu

Genetic differences have been suggested as a possible cause for variation in responses to exogenous hormones. Here we evaluated the effect of follicle ablation, exogenous FSH and coasting time prior to ovum pick-up (OPU) on the number of follicles suitable for aspiration, oocyte quality, and cleavage rate in Ethiopian Boran cows. The experiment was carried out in three parts, I) Cows were synchronized using 500µg PGF2α given 11 days apart. Cows were then subjected to a biweekly ovum pickup session before ovulation (n=5) or starting Day 7 after ovulation (n=4) for three weeks. II) Cows were similarly synchronized and all visible follicles were ablated on the first days of overt estrus which were then further grouped into cows that received a divided dose of 350IU FSH (n=5) or 175IU FSH (n=5) over three days. In both groups OPU was carried out weekly starting 48h after the last FSH for six weeks. III) A similar protocol as in part II was carried out but coasting period was increased to 72hrs for cows that received 350IU FSH as divided dose (n=5) and 48hrs coasting period for single 350IU FSH dose (n=5). The covariates of follicles and oocyte were not affected (P>0.05) by corpus luteum presence at OPU. The mean number of medium (7.36±0.57) and large (8.28±0.96) follicles were significantly higher (P<0.05) in group that received divided 350IU FSH. Similarly, the mean number of Grade-1 (4.19±0.24) and Grade-2 (4.32±.27) COC, maturation rate (70.41%) and cleavage rate (47.5%) were significantly higher (P<0.05) in group that received 350IU FSH. COC quality was significantly (P<0.05) influenced by costing period. However, both maturation and cleavage rates were not affected by the coasting period. This study demonstrated that follicular ablation and treatment with FSH improves follicular population and oocyte recovery rate in Boran cows.


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