122 THE SURVIVAL RATE AND EMBRYONIC QUALITY OF BOVINE PARTHENOGENETIC BLASTOCYSTS POST-VITRIFICATION CRYOPRESERVATION

2010 ◽  
Vol 22 (1) ◽  
pp. 220
Author(s):  
P. Zhang ◽  
Z. W. Wang ◽  
B. Tang ◽  
J. B. Zhang ◽  
Z. Y. Li
Keyword(s):  
Survival Rate ◽  
Sucrose Solution ◽  
Polar Body ◽  
Cell Counting ◽  
Differential Staining ◽  
Hatching Rate ◽  
First Polar Body ◽  
Significant Difference

Cryopreservation of oocytes and embryos is very useful to conservation of animal genetic resources. Recently, parthenogenesis has received considerable attention as a tool for the production of stem cells. Oocytes and embryos undergo considerable morphological changes and functional damage during cryopreservation, and the survival rate is highly depending on species and developmental stage of oocytes and embryos. The aim of this study was to investigate the survival rate and embryonic quality of bovine parthenogenetic blastocysts post-vitrification cryopreservation. Cumulus-oocyte complexes (COC) from slaughterhouse ovaries were aspirated from 2-mm to 8-mm visible follicles with a 5-mL syringe. The COC were matured in vitro for 22 h in bicarbonate-buffered TCM199 media supplemented with 1 mg mL-1 of FSH, 10 mg mL-1 of LH, 1 mg mL-1 of 17-βiestradiol, and 10% FBS. After in vitro maturation, cumulus cells were removed from COC, oocytes with first polar body were activated by 5 μM ionomycin for 5 min and 2 mM 6-DMAP for 4 h. Subsequently, oocytes were co-cultured with bovine fetal fibroblast cells in SOF media supplemented with amino acids (1% NEAA and 2% EAA), 4 mg mL-1 of BSA, and 10% FBS at conditions of 38.5°C and 5% CO2 for 7 to 9 days. The good expanded blastocysts were selected and refrigerated in different vectors [glass micropipettes (GMP) and straws] and same vitrification solution (VS, 20% EG + 20% DMSO). Blastocysts were exposed to VS, loaded on vectors, and plunged into liquid nitrogen within 25 s. After two days refrigeration, vitrified blastocysts were thawed in air for 10 s and placed into 0.25 M sucrose solution for 1 min and 0.15 M sucrose solution for 5 min. Then, the blastocysts were cultured in the SOF medium same as above. Our results showed that when VS was 20% EG + 20% DMSO, the hatching rate (65%) of blastocysts loaded into GMP was significantly higher (P < 0.05) than that (19%) of blastocysts loaded into straws post-vitrification. Meanwhile, vitrified and nonvitrified blastocysts were fixed and stained for differential cell counting as described by Thouas GA et al. 2001 Reprod. Biomed. Online 3, 25-29). By the differential staining, the total cells of nonvitrified parthenogenetic bovine blastocysts were 102.7, which was higher (P > 0.05) than that (86.7) of vitrified blastocysts. Also, no significant difference (P > 0.05) was seen on ratios between vitrified blastocysts (ICM/TE = 0.22) and nonvitrified blastocysts (ICM/TE = 0.25). Our results indicated that a glass micropipette vector was much better than a straw in vitrification cryopreservation of bovine parthenogenetic blastocysts and caused less damage to blastocyst cells. This study lays the foundation for further research to increase the survival rate of vitrification cryopreservation of bovine embryos. This work was supported by the grant from national support plan, China, No. 2007BAD55B03; corresponding author: Ziyi Li.

scholarly journals 298RED DEER (CERVUS ELAPHUS) PARTHENOGENETIC BLASTOCYSTS PRODUCED USING IONOMYCIN/6DMAP ACTIVATION

2004 ◽  
Vol 16 (2) ◽  
pp. 268 ◽  
Author(s):  
S.E. Beaumont ◽  
D.K. Berg ◽  
G.W. Asher
Keyword(s):  
Red Deer ◽  
Polar Body ◽  
Negative Control ◽  
Blastocyst Development ◽  
Cell Counts ◽  
First Polar Body ◽  
Significant Difference ◽  
Chi Square Analysis ◽  
Polar Body Extrusion

Successful activation of red deer oocytes is a necessary prerequisite for the cloning of red deer individuals with desirable genetic characteristics. To investigate this, an established biphasic protocol used for oocyte activation in sheep was investigated for suitability. The method chosen was 5μM Ionomycin for 5min followed by 2mM 6DMAP for 3h ( Loi P et al., 1998 Biol. Reprod. 58, 1177–1187). The medium used during activation and subsequent culture was Deer Synthetic Oviduct Fluid, which has been shown to support routine in vitro fertilization and blastocyst development (15%) of in vitro-matured red deer oocytes (DSOF, Berg D et al., 2003 Theriogenology 59, 189–205). Red deer abattoir-derived COCs were matured in vitro for 22h before random allocation across 3 treatment groups comprising a standard IVF group, the activation group and a negative control group exposed to medium only. Activation treatment oocytes were stripped of cumulus by vortexing in 0.1% hyaluronidase before selecting for first polar body extrusion. First-step activation was performed in medium comprising HEPES-buffered IVF-DSOF containing 4mM Ca2+. Second-step activation used 3mM Ca2+ early DSOF under 7% O2, 5% CO2, and 88% N2 at 38.5°C. Standard IVF was conducted at 23h post-IVM using 4mM Ca2+ IVF-DSOF and 0.5×106mL−1 final sperm concentration. Following activation and IVF, oocytes were washed 3 times in HEPES DSOF before culture for 7 days in sequential DSOF with late DSOF on Day 4 containing 1.5mM Ca2+. Cleavage was assessed 24h after activation, and all blastocysts were fixed for cell counts. Four replicates of each treatment were performed. Cleavage and blastocyst rates were examined by chi-square analysis and cell numbers by ANOVA. First polar body extrusion rate was 84%. Cleavage was similar between the activation treatment and IVF (P&gt;0.05 ); but a significant difference was found in blastocyst development rates (P&lt;0.05) with the Ionomycin and 6DMAP protocol being superior to the IVF treatment. Exposure to high Ca2+ media alone resulted in only 5% of the negative control oocytes cleaving to 2 cells. Results show that Ionomycin and 6DMAP are effective in activating red deer oocytes and DSOF is a suitable medium to produce parthenogenetic blastocysts.

scholarly journals REPRODUCTIVE PERFORMANCE OF DOMESTICATED BROODSTOCK OF SILVER PERCH, Bidyanus bidyanus (MITCHELL 1838) AND THE RELATIONSHIP BETWEEN OIL GLOBULE FRAGMENTATION AND EGG QUALITY

2017 ◽  
Vol 12 (2) ◽  
pp. 43 ◽  
Author(s):  
Sulaeman Sulaeman ◽  
Ravi Fotedar
Keyword(s):  
Survival Rate ◽  
Reproductive Performance ◽  
Egg Quality ◽  
Fertilization Rate ◽  
Hatching Rate ◽  
High Fecundity ◽  
Fertilisation Rate ◽  
Significant Difference ◽  
The Relationship

The experiments investigated the reproductive performance of the domesticated broodstock of the silver perch and the relationship between various degrees of oil globule fragmentation and egg quality. Six years old of second generation broodstock (n=3) were evaluated based on the fecundity, fertilisation rate, hatching rate, the degree of oil fragmentation of egg, and the quality of embryos and larvae produced. The fragmentation were grouped into three categories: un-fragmented (cat-1), moderately fragmented (cat-2), and highly fragmented (cat-3). The results showed that the broodstock had a relatively high fecundity (132,400 ± 7,22), fertilization rate (94.27 ± 1.28%), and hatching rates (87.94 ± 1.23%). The survival rate of larvae at 12 days post hatching (dph) in cat-1 (71.3 ± 0.9%) was higher than cat-2 (66.7 ± 0.9%) whereas cat-2 was higher than cat-3 (61.3 ± 0.3%). The eggs was dominated by cat-1 (78.11 ± 2.44%) which was significantly higher than cat-2 (21.26 ± 2.45%) and cat-3 ones (0.40 ± 0.21%). The survival rate of embryo at 20 hours post spawning (hps) and hatching rate of cat-1 (95.33 ± 0.00% and 93.33 ± 0.00%) and cat-2 (90.00 ± 0.00% and 85.00 ± 0.00%) were significantly higher than cat-3 (72.33 ± 1.76% and 60.33 ± 0.00%). The total length (TL) of the larvae of cat-1 and cat-2 (8.44 ± 0.21 mm and 8.35 ± 0.23 mm respectively) were significantly higher than larvae of cat-3 (7.09 ± 0.14 mm). No significant difference was found in the larval deformities among any categories. In conclusion, the reproductive performance of six year-old broodstock silver perch showing acceptable performance and egg categorisation based on oil globule fragmentation can be used as a useful tool to indicate eggs quality of silver perch.

scholarly journals The Role of Ca2 + in Maturation and Reprogramming of Bovine Oocytes: A System Study of Low-Calcium Model

2021 ◽  
Vol 9 ◽  
Author(s):  
Lin Meng ◽  
Hongmei Hu ◽  
Zhiqiang Liu ◽  
Luyao Zhang ◽  
Qingrui Zhuan ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Polar Body ◽  
Rna Seq ◽  
Low Calcium ◽  
Maturation Medium ◽  
First Polar Body ◽  
Significant Difference ◽  
Bovine Oocytes

[Ca2+]i is essential for mammalian oocyte maturation and early embryonic development, as those processes are Ca2+ dependent. In the present study, we investigated the effect of [Ca2+]i on in vitro maturation and reprogramming of oocytes in a lower calcium model of oocyte at metaphase II (MII) stage, which was established by adding cell-permeant Ca2+ chelator BAPTA-AM to the maturation medium. Results showed that the extrusion of the first polar body (PB1) was delayed, and oocyte cytoplasmic maturation, including mitochondrial and endoplasmic reticulum distribution, was impaired in lower calcium model. The low-calcium-model oocytes presented a poor developmental phenotype of somatic cell nuclear transfer (SCNT) embryos at the beginning of activation of zygotic genome. At the same time, oxidative stress and apoptosis were observed in the low-calcium-model oocytes; subsequently, an RNA-seq analysis of the lower-calcium-model oocytes screened 24 genes responsible for the poor oocyte reprogramming, and six genes (ID1, SOX2, DPPA3, ASF1A, MSL3, and KDM6B) were identified by quantitative PCR. Analyzing the expression of these genes is helpful to elucidate the mechanisms of [Ca2+]i regulating oocyte reprogramming. The most significant difference gene in this enriched item was ID1. Our results showed that the low calcium might give rise to oxidative stress and apoptosis, resulting in impaired maturation of bovine oocytes and possibly affecting subsequent reprogramming ability through the reduction of ID1.

scholarly journals The Number of LH Receptor could Predict the Success of Oocyte Maturity in the Process of In Vitro Maturation

2016 ◽  
Author(s):  
Adek Amansyah
Keyword(s):  
In Vitro Maturation ◽  
Clinical Laboratory ◽  
Polar Body ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Oocyte Maturity ◽  
Lh Receptor ◽  
First Polar Body ◽  
Significant Difference

Objective: To evaluate the relationship between the number of LH receptor and the success of oocyte maturity in the process of in vitro maturation (IVM). Method: This experimental study was conducted in the Permata Hati Infertility Clinical Laboratory, Dr. Sardjito General Hospital, Yogyakarta, with the samples of 300 oocytes obtained through collecting immature cow’s oocytes from the abattoir and grouped the oocytes into 3 (three) groups based on the pattern of oocyte cumulus cells on the vesicle germinal stage 2 - 8 mm with three layers of cumulus cell. The sample of the cumulus cells from these three groups were taken and the LH receptor examination was done with immunohistochemistry. After that, the IVM process was performed to the three groups and its development for 24 hours was evaluated. Its maturation quality was evaluated with the emergence of the first polar body (1PB) and compared to the other groups and related to the number of LH receptor in the three groups. Result: The result of this study indicated that the oocyte cumulus cells showed a difference of function during IVM process. The maturity rate in this study showed that the number of LH receptor was related to the morphological pattern of oocyte cumulus cells with oocyte maturity. The maturity of the cumulus cells which 100% covered the oocyte was higher than that of the cumulus cells which > 50% and < 30% covered the oocytes, namely, 74% compared to 60% and 12%. The result of this study also showed that the average number of LH receptors in the three groups (A, B, and C) was 183.4, 78.8, and 24.0 respectively. A significant difference was found in the three groups (p < 0.0001). When related to IVM maturity, this difference showed that the bigger number of oocyte cumulus cells influenced the oocyte maturity. Conclusion: The number of LH receptor can be used as a prediction to determine the success of oocyte maturation in the process of in vitro maturation. [Indones J Obstet Gynecol 2013; 1-4:183-7] Keywords: IVM, LH receptor, oocyte cumulus cell

79 A COMBINATION OF ETHYLENE GLYCOL AND PROPYLENE GLYCOL IS SUPERIOR TO INDIVIDUAL CRYOPROTECTANTS FOR THE VITRIFICATION OF IMMATURE PORCINE OOCYTES

2013 ◽  
Vol 25 (1) ◽  
pp. 187 ◽  
Author(s):  
T. Somfai ◽  
K. Kikuchi ◽  
M. Nakai ◽  
M. Kaneda ◽  
S. Akagi ◽  
...  
Keyword(s):  
Survival Rate ◽  
Ethylene Glycol ◽  
Propylene Glycol ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Membrane Integrity ◽  
Control Groups ◽  
Cell Numbers ◽  
First Polar Body

We compared the feasibility of ethylene glycol (EG) and propylene glycol (PG) for the vitrification of immature porcine cumulus–oocyte complexes (COC). Porcine COC collected from 3- to 6-mm follicles of slaughterhouse-derived ovaries were subjected to solid-surface vitrification (Somfai et al. 2010 Theriogenology 73, 147–156) either in 35% (v/v) EG or 35% (v/v) PG or in the mixture of 17.5% (v/v) EG and 17.5% (v/v) PG. After warming, the COC were subjected to in vitro maturation, IVF, and embryo culture according to Kikuchi et al. (2002 Biol. Reprod. 66, 1033–1041). Oocyte survival and maturation rates were assessed after in vitro maturation by evaluating membrane integrity and the extrusion of the first polar body. All live oocytes were subjected to IVF and in vitro culture. Cleavage and blastocyst rates were calculated from the total number of oocytes subjected to IVF on Day 2 (Day 0 = IVF) and Day 7, respectively. Total-cell (blastomeres) numbers in blastocysts were recorded on Day 7 after staining with Hoechst 33342. In Experiment 1, competence parameters of oocytes vitrified either in EG-based (EG group; n = 310) or a PG-based (PG group; n = 265) vitrification media were compared with those in the nonvitrified control (n = 160). The experiment was replicated 4 times. In Experiment 2, the competence parameters of oocytes vitrified with the combination of 17.5% EG and 17.5% PG (EG+PG group; n = 397) were compared with those in nonvitrified control (n = 245) and toxicity control (TC, exposed to cryoprotectants without cooling; n = 245) groups. Five replications were performed. Results were analyzed by ANOVA. Differences with P < 0.05 were considered significant. In Experiment 1, the mean survival rate of vitrified oocytes was significantly higher (P < 0.05) in 35% PG compared with that in 35% EG (73.3 and 25.9%, respectively). Maturation rates of surviving oocytes did not differ among vitrified (PG and EG) and nonvitrified control groups (71.1, 62.4, and 64.0%, respectively). After IVF of surviving oocytes, blastocyst formation rate in the group vitrified in EG was higher (P < 0.05) compared with that vitrified in PG but was lower (P < 0.05) compared with control (10.8, 2.0, and 25.0%, respectively). Mean cell numbers in blastocysts did not differ among EG, PG, and control groups (50.5, 47.7, and 48.7, respectively). In Experiment 2, survival of immature oocytes in the EG+PG group was 42.6%. After IVF, 10.7% of oocytes developed to the blastocyst stage in the EG+PG group, which was lower (P < 0.05) than those of the control (18.1%) and TC (23.3%) groups. Blastocyst rates in the control and TC groups were not statistically different. Mean cell numbers in blastocysts did not differ significantly among the EG+PG, control, and TC groups (61.6, 59.3, and 53.3, respectively). In conclusion, 35% PG provided a higher oocyte survival rate after vitrification compared with 35% EG. However, presumably due to toxic effects, 35% PG greatly reduced the development competence of oocytes. The combination of 17.5% EG and 17.5% PG yielded higher survival rates than did 35% EG, without any toxic effect on oocytes.

scholarly journals ID: 1066 Effects of biotin supplementation on the in vitro maturation of oocytes and the development of parthenogenetic diploid porcine embryos

2017 ◽  
Vol 4 (S) ◽  
pp. 148
Author(s):  
Nguyen Ba Tu ◽  
Bui Hong Thuy ◽  
Nguyen Van Thuan
Keyword(s):  
In Vitro Maturation ◽  
Polar Body ◽  
Biotin Deficiency ◽  
Preimplantation Stage ◽  
Maturation Medium ◽  
First Polar Body ◽  
Porcine Oocyte ◽  
Biotin Supplementation

In mammals, Biotin serves a coenzyme in the metabolism of glucose, amino acids, and fatty acids. Biotin deficiency causes decreased rates of cell proliferation, disfunction in germ cells and fetal development. This study was carried out to determine the influence of Biotin supplementation to invitro maturation medium on the development of porcine oocyte and embryos. Biotin (0.0, 1.0, 10.0, 100.0 mg/l, respectively) was added into the oocyte maturation medium, the quality of mature oocytes was evaluated after 42h culturing. The parthenogenetic diploid embryos were produced by using electro-activation system, the quality of embryos was noted at 1-4 cells stage. The results showed that, Biotin can enhance the formation of the first polar body at the concentration of 10 mg/l, it can also improve the activation efficiency of parthenogenetic diploid embryos at the preimplantation stage from 2-4 cells. Therefore, the supplementation of 10mg/l Biotin to the in-vitro maturation medium has a beneficial effect on the parthenogenetic diploid embryos development in the pig.

70 EFFECT OF CRYOPROTECTANT CONCENTRATION ON THE IN VITRO SURVIVAL AND CELL PROLIFERATION OF PORCINE BLASTOCYSTS VITRIFIED USING THE OPEN PULLED STRAW SYSTEM

2008 ◽  
Vol 20 (1) ◽  
pp. 116
Author(s):  
C. Cuello ◽  
J. Sanchez-Osorio ◽  
C. Almiñana ◽  
M. A. Gil ◽  
I. Caballero ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Survival Rate ◽  
Nuclear Antigen ◽  
Proliferation Index ◽  
Hatching Rate ◽  
Hoechst 33342 ◽  
Number Range ◽  
Significant Difference ◽  
In Vitro Survival

The objective of the present project was to study the effect of the concentrations of ethylene glycol (EG) and dimethyl sulfoxide (DMSO) during vitrification on the survival and hatching rates of porcine blastocysts. Embryos were collected by laparotomy from weaned crossbred sows (n = 18), vitrified, and warmed (one-step dilution) as described by Cuello et al. (2004 Theriogenology 62, 1144–1152). Vitrification was performed in different concentrations of EG and DMSO (15%, 16%, and 17% v/v for each cryoprotectant) or in an EG-based medium (40% v/v) using superfine open pulled straws. Fresh and vitrified blastocysts were cultured for 24 h in TCM199 and assessed by stereomicroscopy during the culture. Blastocysts that reformed their blastocoelic cavities after warming, displaying a normal zona pellucida and excellent appearance, were considered viable. The in vitro survival rate was defined as the ratio of viable embryos divided by the total number of embryos cultured. The hatching rate is determined as the ratio of the number of embryos hatched in vitro to the total number embryos cultured. Some vitrified and fresh embryos classified as viable were processed for Hoechst 33342 staining and proliferation cell nuclear antigen (PCNA) inmunolocalization. The proliferation index was defined as the number of PCNA-positive nuclei divided by the total number of nuclei stained with Hoechst 33342. Data were analyzed by ANOVA using the MIXED procedure (SPSS version 11.5; SPSS, Inc., Chicago, IL, USA). Data were expressed as mean values � SEM. The survival rate was similar for fresh and vitrified blastocysts, except for blastocysts vitrified using 15% cryoprotectants, which displayed a lower (P < 0.05) survival rate (84.2 � 4.8%) than fresh blastocysts (94.6 � 5%) and blastocysts vitrified using 40% EG, 16%, or 17% EG-DMSO (88.8 � 4.9, 96.8 � 4.9, and 96.4 � 5%, respectively). Fresh blastocysts showed a higher (P < 0.05) hatching rate (80.7 � 4.5%) than their vitrified counterparts (range: 48.4 � 7.7–55.3 � 7.8%). Vitrified and fresh blastocysts showed similar cell proliferation indexes (range: 75.8 � 3.2–83.7 � 3). When only hatched blastocysts among groups were compared, the proliferation rate decreased (P < 0.05) after vitrification with 17% EG-DMSO. Among vitrification groups, there was no significant difference in the number of total cells. However, vitrified blastocysts had a lower (P < 0.05) total cell number (range: 116.6 � 6.7–124.8 � 6.6) than fresh blastocysts (195.5 � 11.4). In conclusion, under our experimental conditions, the concentration of EG-DMSO can be decreased until 16% in the vitrification medium with no reduction of the in vitro developmental ability of the blastocysts. In addition, a 40% EG-based medium can be used for vitrification with results similar to those achieved using a medium containing 16% EG-DMSO.

174 EFFECT OF HUMAN ENDOTHELIAL PROGENITOR CELLS ON IN VITRO MATURATION OF PORCINE OOCYTES AND PARTHENOGENETIC EMBRYO DEVELOPMENT COMPETENCE

2017 ◽  
Vol 29 (1) ◽  
pp. 195
Author(s):  
S. H. Lee ◽  
H. J. Oh ◽  
M. J. Kim ◽  
G. A. Kim ◽  
E. M. N. Setyawan ◽  
...  
Keyword(s):  
Growth Factor ◽  
Embryo Development ◽  
Oocyte Maturation ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Polar Body ◽  
Blastocyst Formation ◽  
First Polar Body ◽  
Significant Difference

In oocyte maturation, hepatocyte growth factor and vascular endothelial growth factor (VEGF) contribute to promote granulosa cell proliferation and cumulus cell expansion. It is well known that human endothelial progenitor cells (hEPC), which are isolated from monocytes and macrophages, secrete a variety of growth factors, such as hepatocyte growth factor and VEGF, and improve the process of angiogenesis. Therefore, the aim of this study was to investigate the effects of hEPC on in vitro oocyte maturation and subsequent embryo development in pigs. To isolate and culture hEPC, human peripheral blood sample was collected from a healthy donor and peripheral blood mononuclear cells were separated. The peripheral blood mononuclear cells were seeded into flask with defined Keratinocyte-SFM-based medium and incubated at 37°C, 5% CO2. The hEPC were cultured and cryopreserved until use for co-culturing with porcine oocytes obtained from a local slaughterhouse ovaries. Cumulus-oocyte complexes were randomly cultured in 2 groups; 1) co-culturing with hEPC and 2) culturing without hEPC. Cumulus-oocyte complexes were cultured in the in vitro maturation (IVM) medium containing TCM-199 supplemented with 0.57 mM cysteine, 0.91 mM sodium pyruvate, 5 μL mL−1 of insulin-transferrin-selenium solution 100X (Invitrogen, Seoul, South Korea), 10% porcine follicular fluid, 10 IU mL−1 of eCG, and 10 IU mL−1 of hCG. After IVM, the first polar body extrusion was observed under the microscope. To evaluate embryo development competence, the matured oocytes were activated with electrical stimulus and cultured in porcine zygote medium-5 for 7 days. The cleavage and blastocyst formation rates were observed on Day 2 and 7, respectively. Also, blastocysts were stained with Hoechst 33342 and total blastocyst cell numbers were evaluated under a fluorescence microscope. As a result, the oocyte maturation rate or first polar body extrusion rate of the hEPC co-culture group (90.06 ± 0.75) was significantly higher than the control group (90.06 ± 0.75 v. 85.79 ± 0.59; P < 0.05). There was no significant difference between the hEPC co-cultured and the control groups in cleavage rate. However, a significant difference in blastocyst formation rate was observed between the hEPC co-cultured and the control groups (28.45 ± 4.92 v. 15.87 ± 2.27; P < 0.05), whereas total blastocyst cell numbers did not show significant difference between the 2 groups. The all data were analysed by unpaired t-test using GraphPad Prism 5.0 (GraphPad Software Inc., La Jolla, CA, USA). Values are means ± standard error of mean. In conclusion, the results in the present study demonstrated that co-culturing with hEPC improved the in vitro oocyte maturation and blastocyst formation rate. Also, we are underway in analysing the concentration of VEGF families in the hEPC co-culture medium after IVM. For further study, we will analyse the genes of the VEGF signaling pathway in the cumulus cells and matured oocytes derived from the 2 groups. This research was supported by Nature Cell (#550-20150030), global PH.D Fellowship Program through NRF funded by the Ministry of Education (NRF-20142A1021187), and Research Institute for Veterinary Science, the BK21 plus program.

scholarly journals In Vitro Oocyte Maturation Rate of Mice with Polycystic Ovarian Syndrome in the Presence of α-Linolenic Acid Antioxidant

2020 ◽  
Vol 22 (2) ◽  
Author(s):  
Amaneh Moradi ◽  
Fatemeh Ghasemian ◽  
Farhad Mashayekhi
Keyword(s):  
Linolenic Acid ◽  
In Vitro Maturation ◽  
Polycystic Ovary ◽  
Polar Body ◽  
Reproductive Age ◽  
Estradiol Valerate ◽  
Control Group ◽  
First Polar Body

Background: One of the most common endocrine and metabolic disorders is Polycystic ovary syndrome (PCOS), which has been reported in about 10% of women during the reproductive age. Objectives: This study was designed to investigate the efficiency of α-Linolenic acid (ALA) on in vitro maturation (IVM) and the quality of mouse oocytes with PCOS. Methods: Female NMRI mice (30 - 35 day-old) were developed by the injection of 4 mg estradiol valerate dissolved in 0.2 mg sesame oil for 60 consecutive days. In the following, the PCOS ovaries were dissected and oocytes were cultured in the maturation medium supplemented with different dosages of α-linolenic acid (0, 50, 100 µM). The presence of the first polar body was considered the sign of the nuclear maturation of the oocyte. The expression of mitochondrial transcription factor A (TFAM) gene in mature oocytes was investigated by Quantitative Real-time PCR. Results: The in vitro maturation and TFAM gene expression rates of PCOS oocytes in the medium treated with 50 µM of ALA (84 ± 7.9 and 0.46 ± 0.09, respectively) were significantly higher than the control group (P < 0.05). Conclusions: The ALA could improve the IVM rate and quality of PCOS oocytes by higher expression of TFAM gene.

scholarly journals The Effectiveness of Candidate Probiotic Bacteria to Control Vibriosis Disease

2017 ◽  
Vol 5 (2) ◽  
pp. 1
Author(s):  
Mulyati Mulyati ◽  
Suryati Suryati ◽  
Irfani Baga
Keyword(s):  
Survival Rate ◽  
Sea Water ◽  
Challenge Test ◽  
Probiotic Bacteria ◽  
Pathogenicity Test ◽  
Inhibitory Ability ◽  
Significant Difference ◽  
Portunid Crab

The study aims to isolate, characterize, and examine probiotic bacteria's inhibitory ability against Vibrio harveyi bacteria, both in-vitro and in vivo. Methods used in the study consist of 1) An Isolation of Candidate Probiotic Bacteria, 2) An Antagonistic Test of Candidate Probiotic Bacteria in vitro, 3) An Identification of Bacteria, 4) A Pathogenicity Test of Candidate Probiotic Bacteria, 5) An Antagonistic Test of Candidate Probiotic Bacteria against V. harveyi in vivo. According to the isolation of candidate probiotic bacteria, there are 18 isolated candidate probiotic. After being tested for its inhibitory ability in vitro, there are 8 isolates with zone of inhibition as follows: isolate MM 7 from intestine (22 mm), isolate MM 6 from intestine (12 mm), isolate MM 10 from sea water (10 mm), isolate MM 5 from intestine (9 mm), isolate MM 4 from intestine (8 mm), isolate MM 3 from intestine (7 mm), isolate MM 2.2 from intestine (7 mm), isolate MM 2.1 from intestine (7 mm). Eight genera of the candidate probiotic bacteria is derived from Portunid crab, they are Staphylococcus, Streptococcus, bacillus, vibrio, Alcaligenes, Lactobacillus, micrococcus. Before proceeding the V. harveyi bacterial challenge test in vivo, three potential isolates consisting of MM6, MM7 and MM10 as the probiotic bacteria are pathogenicity-tested against V. harveyi. The survival rate of Portunid crab on pathogenicity test using MM6, MM7 and MM10 generates 91.11-100%, while the control generates 100% survival rate. Variance analysis result through post-hoc Tukey's Honest Significant Difference (HSD) test at 95% confidence interval indicates that isolate MM7 and MM10 are significantly able to increase hatchling Portunid crab's survival rate.

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