scholarly journals Reduced Expression of Interleukin-11 and Interleukin-6 in the Periimplantation Endometrium of Excessive Ovarian Responders during in Vitro Fertilization Treatment

2006 ◽  
Vol 91 (8) ◽  
pp. 3181-3188 ◽  
Author(s):  
Guneet Makkar ◽  
Ernest H. Y. Ng ◽  
William S. B. Yeung ◽  
P. C. Ho
Keyword(s):  
High Serum ◽  
Th2 Cytokines ◽  
Vitro Fertilization ◽  
Negative Effect ◽  
Group A ◽  
Uterine Flushing ◽  
Group B ◽  
Natural Cycles

Abstract Context: Impaired implantation in assisted reproduction cycles with high serum estradiol (E2) concentrations may be related to abnormal endometrial functions. Objective: The in vivo expression of T helper type 2 (Th2) cytokines in the periimplantation endometrium of infertile patients was compared between natural and stimulated cycles. Interventions and Main Outcome Measures: Uterine flushings and endometrial biopsies were collected 7 d after the LH surge in natural cycles or after human chorionic gonadotropin injection in stimulated cycles. Th2 cytokines were determined by immunolocalization and by ELISA. Natural cycles were in group A, whereas stimulated cycles with peak serum E2 of no more than 20,000 pmol/liter (moderate responders) and more than 20,000 pmol/liter (excessive responders) were classified as group B and group C, respectively. Results: Higher E2 had a negative effect on IL-11 and IL-6 expression in the endometrium and IL-11 concentration in the uterine flushing. In endometrial biopsies, a significantly lower immunostaining of stromal IL-11 (P < 0.001) and glandular IL-6 (P < 0.05) was detected in group C compared with that of groups A and B. IL-11 concentration by ELISA was significantly lower in group C (P < 0.05). Endometrial leukemia inhibitory factor and IL-4 expression was similar in the three groups. In uterine flushings, a significantly higher percentage of women in group C had undetectable IL-11 and a lower IL-11 concentration (P < 0.01) compared with group A, whereas no difference in IL-6 concentration was noted in the three groups. Conclusion: Reduced expression of IL-11 and IL-6 in periimplantation endometrium may account for lower implantation in excessive responders.

scholarly journals High Serum FSH is Associated with Brown Oocyte Formation and a Lower Pregnacy Rate in Human IVF Parctice

2016 ◽  
Vol 39 (2) ◽  
pp. 677-684 ◽  
Author(s):  
Hongyi Xu ◽  
Kai Deng ◽  
Qingbing Luo ◽  
Juan Chen ◽  
Xin Zhang ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Embryo Quality ◽  
High Serum ◽  
Vitro Fertilization ◽  
Good Quality Embryo ◽  
Serum Hormone ◽  
Group A ◽  
Group B ◽  
Ivf Et

Background/Aims: To investigate whether brown zona pellucida (ZP) of oocytes affects the outcome of fertilization, embryo quality and pregnancy rate in in vitro fertilization-embryo transfer (IVF-ET). Methods: Based on the ZP color of their oocytes, a total number of 703 patients dated from 2012 to 2014 were divided into a normal egg group (group A) and a brown oocyte group (group B), with 629 and 74 cases, respectively. Clinical characteristics, gonadotropin (Gn) days, Gn dosage, serum hormone levels on the day of human chorionic gonadotropin (HCG) injection, ZP thickness (ZPT) of the eggs, fertilization rate, rescue intracytoplasmic sperm injection (rICSI) rate, good-quality embryo rate and pregnancy rate were compared between the two groups. Results: No significant differences were found in the duration and the causes of infertility, and their basal level of endocrine hormone before IVF-ET between normal egg group and brown egg group. The level of serum hormone including estradiol, progesterone and luteinizing hormone on the day of HCG injection were again similar. Moreover, there were no differences in number of mature oocytes, oocyte fertilization rates and rICSI rates after IVF between the two groups. However, we observed that the ZPT of brown oocytes (group B) was higher than that of normal oocytes (group A). Moreover, the Gn dosage and FSH levels on the day of HCG injection were significantly higher in group B than in group A and the good-quality embryo rate and pregnancy rate in group B were lower than those in group A. Conclusion: Compared with normal eggs, oocytes with a brown ZP were found to have a higher ZPT, lower embryo quality and lower pregnancy rate, which might be due to a high Gn dosage injection and high serum FSH levels during IVT-ET cycles.

Salivary oestradiol and progesterone after in vitro fertilization and embryo transfer using different luteal support regimens

1990 ◽  
Vol 2 (4) ◽  
pp. 351 ◽  
Author(s):  
YF Wong ◽  
EP Loong ◽  
KR Mao ◽  
PP Tam ◽  
NS Panesar ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Transfer ◽  
Luteal Phase ◽  
Biologically Active ◽  
Luteal Support ◽  
Vitro Fertilization ◽  
Group A ◽  
Group B ◽  
Ivf Et

Salivary oestradiol (E2) and progesterone (P) levels have been shown to reflect the biologically active fractions in the serum. The luteal-phase status of stimulated cycles was investigated after in vitro fertilization and embryo transfer (IVF-ET). Thirty patients were randomly allocated to one of three luteal therapy groups: group A had no support, group B had intramuscular P and group C had intramuscular P and human chorionic gonadotrophin (hCG). One pregnancy was achieved in group A, two in group B and three in group C. Significant correlations between salivary and serum levels of E2 and of P in matched samples during luteal phase were found. Salivary E2 levels from luteal day 8 through day 14 and P levels from day 3 through day 14 were significantly higher in the pregnant than in the nonpregnant cycles. Among the nonpregnant cycles, salivary E2 and P levels were significantly higher in group C than in group A or B. These findings suggest that, in stimulated cycles for IVF-ET, determination of salivary E2 and P levels may be used as reliable alternatives to serum concentrations for assessing the luteal phase. Also, the additional hCG has an enhanced luteotrophic effect, as reflected by the higher salivary E2 and P levels, which may lead to a better pregnancy rate.

scholarly journals Photopolymerizable Injectable Cartilage Mimetic Hydrogel for the Treatment of Focal Chondral Lesions: A Proof of Concept Study in a Rabbit Animal Model

2018 ◽  
Vol 47 (1) ◽  
pp. 212-221 ◽  
Author(s):  
Cecilia Pascual-Garrido ◽  
Elizabeth A. Aisenbrey ◽  
Francisco Rodriguez-Fontan ◽  
Karin A. Payne ◽  
Stephanie J. Bryant ◽  
...  
Keyword(s):  
Animal Model ◽  
Chondrogenic Differentiation ◽  
Osteochondral Lesions ◽  
Chondral Defect ◽  
Group A ◽  
Group B ◽  
Safranin O

Background: In this study, we investigate the in vitro and in vivo chondrogenic capacity of a novel photopolymerizable cartilage mimetic hydrogel, enhanced with extracellular matrix analogs, for cartilage regeneration. Purpose: To (1) determine whether mesenchymal stem cells (MSCs) embedded in a novel cartilage mimetic hydrogel support in vitro chondrogenesis, (2) demonstrate that the proposed hydrogel can be delivered in situ in a critical chondral defect in a rabbit model, and (3) determine whether the hydrogel with or without MSCs supports in vivo chondrogenesis in a critical chondral defect. Study Design: Controlled laboratory study. Methods: Rabbit bone marrow–derived MSCs were isolated, expanded, encapsulated in the hydrogel, and cultured in chondrogenic differentiation medium for 9 weeks. Compressive modulus was evaluated at day 1 and at weeks 3, 6, and 9. Chondrogenic differentiation was investigated via quantitative polymerase reaction, safranin-O staining, and immunofluorescence. In vivo, a 3 mm–wide × 2-mm-deep chondral defect was created bilaterally on the knee trochlea of 10 rabbits. Each animal had 1 defect randomly assigned to be treated with hydrogel with or without MSCs, and the contralateral knee was left untreated. Hence, each rabbit served as its own matched control. Three groups were established: group A, hydrogel (n = 5); group B, hydrogel with MSCs (n = 5); and group C, control (n = 10). Repair tissue was evaluated at 6 months after intervention. Results: In vitro, chondrogenesis and the degradable behavior of the hydrogel by MSCs were confirmed. In vivo, the hydrogel could be delivered intraoperatively in a sterile manner. Overall, the hydrogel group had the highest scores on the modified O’Driscoll scoring system (group A, 17.4 ± 4.7; group B, 13 ± 3; group C, 16.7 ± 2.9) ( P = .11) and showed higher safranin-O staining (group A, 49.4% ± 20%; group B, 25.8% ± 16.4%; group C, 36.9% ± 25.2%) ( P = .27), although significance was not detected for either parameter. Conclusion: This study provides the first evidence of the ability to photopolymerize this novel hydrogel in situ and assess its ability to provide chondrogenic cues for cartilage repair in a small animal model. In vitro chondrogenesis was evident when MSCs were encapsulated in the hydrogel. Clinical Relevance: Cartilage mimetic hydrogel may offer a tissue engineering approach for the treatment of osteochondral lesions.

Effects of Removal of the Statoacoustic Ganglion Complex upon the Growing Otocyst

1976 ◽  
Vol 85 (6_suppl) ◽  
pp. 2-32 ◽  
Author(s):  
Thomas R. Van De Water
Keyword(s):  
Organ Culture ◽  
Inner Ear ◽  
Nerve Fibers ◽  
Trophic Effect ◽  
Group A ◽  
Sensory Structures ◽  
Statoacoustic Ganglion ◽  
Group B

An experiment was designed to answer the question as to whether or not the neural elements of the statoacoustic ganglion complex have a trophic effect upon the histodifferentiation of the sensory structures of the embryonic mouse inner ear anlage as it develops in vitro. The embryonic inner ear anlage with associated otic mesenchyme and statoacoustic ganglion complex was excised from 11, 12, and 13-day CBA/C57 mouse embryos. The inner ear explants of each gestational age group were further divided into two groups: the first group “A” (with) statoacoustic ganglion was explanted to the organ culture system without further surgical intervention; the second group “B” (without) statoacoustic ganglion underwent further surgical manipulation during which their statoacoustic ganglion complexes were dissected away prior to explantation to in vitro. The explanted embryonic inner ears were allowed to develop in organ culture until the equivalent of gestation day 21 in vivo was reached for each group; then all cultures were fixed and histologically processed and stained by a nerve fiber stain, in combination with a stain for glucoprotein membranes. Each specimen was code labeled and scored for histodifferentiation of sensory structures. Light microscopic observations confirmed that in group “A” cultures, statoacoustic ganglion neurons and their nerve fibers were present in association with the developed sensory structures; neither ganglion cell neurons nor their nerve fibers were found to be present in the sensory structures that developed in the group “B” organ culture specimens. Quantification revealed no consistent trend of greater occurrence of any sensory structure in the groups of explants analyzed. The presence of such a trend would have signified the probable existence of a trophic effect of the statoacoustic ganglion neural elements upon development of inner ear sensory structures in the group “A” explants of the 11, 12, and 13-day embryo inner ear organ culture specimens when compared to the aganglionic group “B” cultures. Microscopic comparison of the sensory structures and their sensory hair cells that developed in the organ cultures revealed no differences in the quality of the histodifferentiation of either group “A” or group “B” explants. A base to apex pattern of histodifferentiation of the organ of Corti sensory structures, which has been described to occur in vivo, was noted to occur in the in vitro developed cochlear ducts of all of the explanted inner ears without respect to whether neural elements were present (“A”) or absent (“B”) during development. It was concluded from the quantification of histodifferentiation data and the above observation on the pattern of differentiation of Corti's organ that no trophic effect of neural elements of the statoacoustic ganglion complex influencing the histodifferentiation of sensory structures of 11, 12, and 13-gestation day mouse embryo inner ear explants as they differentiate in vitro could be demonstrated.

Mycophenolate Acid Has a Negative Effect on the Maturation and Immune Function of the Murine Bone Marrow-Derived Dendritic Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3808-3808
Author(s):  
Zhen Cai ◽  
Wenye Huang ◽  
Wenji Sun
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Dendritic Cells ◽  
Immune Function ◽  
De Novo ◽  
Th2 Cytokines ◽  
Negative Effect ◽  
Antigen Presenting

Abstract Mycophenolate mofetil (MMF) is a newly developed immunosuppressor, currently widely used in allogeneic bone marrow transplantation. Its active metabolite, mycophenolic acid (MPA) is a noncompetitive, reversible inhibitor of the enzyme inosine 59-monophosphate dehydrogenase, which plays a major role in the de novo synthesis of guanosine nucleotides. Unlike other cells that also use the salvage pathway for purine biosynthesis, proliferating B and T cells are dependent on the de novo pathway generate guanosine. Thus, MMF exerts its immunosuppressive effects of lymphocyte proliferation. Recently, some studies found that MPA could inhibit the immun immune function of antigen presenting cells. Dendritic cells (DCs), the most potent antigen presenting cells with the unique ability to prime naive T cells, play a central role in antigen processing and presentation to induce T cell response in vitro and in vivo. This study is to evaluate the effects of MPA, the in vivo active metabolite of MMF, on the maturation and immune function of murine bone marrow-derived dendritic cells, and to explore the underlying mechanisms of MMF in graft versus host disease. Bone marrow-derived dendritic cells (DC) were cultured with GM-CSF and IL-4 in the presence of MPA at doses of 0.01 and 0.1μmol/L. The ability of the allostimulatory activities of the DCs on allogeneic T cells was assessed by MLR. IL-12 production in culture supernatant and the Th1/Th2 cytokines such as IL-2, IFN-g, IL-4 and IL-10 levels in mixed lymphocyte reaction (MLR) supernatant were examined by ELISA assays. The activity of NF-κB in DCs was measured with Western blot assays. Our results showed that DCs cultured in the presence of MPA expressed lower levels of CD40, CD80 and CD86, exhibited weaker activity of stimulating the allogeneic T cell proliferation and weaker in antigen presenting function with a concurrent reduction of IL-12 production. MPA-treated DCs stimulated allogeneic T cells to secrete higher levels of Th2 cytokines IL-4 and IL-10 but lower levels of Th1 cytokines IL-2 and IFN-g than did DCs not treated with MPA. The activity of NF-κB was decreased in DCs treated with MPA in a dose-dependent manner. We conclude that MPA, and hence MMF, exerts a negative effect on the maturation and immune function of in vitro cultured DCs, and drives a shift of Th1 cytokines to Th2 cytokines in MLR. This negative effect is associated with a decrease in NF-κB activity. Say something about the significance of this finding regarding GVHD.

scholarly journals Success rate of intrauterine insemination in patients with unknown infertility

2012 ◽  
Vol 69 (4) ◽  
pp. 301-307 ◽  
Author(s):  
Vladimir Jasovic ◽  
Emilija Jasovic-Siveska
Keyword(s):  
Success Rate ◽  
Negative Impact ◽  
Intrauterine Insemination ◽  
Unexplained Infertility ◽  
Mild Form ◽  
Vitro Fertilization ◽  
Group A ◽  
Group B ◽  
Twin Pregnancies

Background/Aim. Unknown cause of infertility exists in 10%-26% of couples with infertility problems. Treatment of these couples depends on the possibility of correcting the unidentified defect over time. Intrauterine insemination (IUI) and ovaluation stimulation are methods of choice in treatment of unexplained fertility, but if a woman is older than 37 years, in vitro fertilization (IVF) could be directly recommended. The aim of this research was to compare the success rate of pregnancies with IUI between the patients with unexplained infertility and the patients with mild form endometriosis. Methods. The study included on 50 patients diagnosed with mild form endometriosis (group A) and 50 patients with unknown cause infertility (group B). Using the same therapeutical protocol, human menopausal gonadothropin (hMG) stimulation and horionic gonadropin (hCG) induction were applied, as well as IUI. Results. The percentage of achieved ovulation was higher in the group B (p < 0.05). During the 3 simulated sequential periods 102 IUI were performed in the group A and 97 IUI in the group B. In the group A there were 6 single and 1 twin pregnancies sucesfully conceived (14%), while in group B there were 9 (18%) single pregnancies. Conclusion. The use of a combination of controled ovarian hyperstimulation and IUI is an effective, cheap and safe method for treating infertility couples, especially couples with unknown cause infertility. Mild form endometriosis, as etiological infertility factor, has a negative impact on IUI success rate.

scholarly journals Pregnancy outcomes of dichorionic triamniotic triplet pregnancies after in vitro fertilization-embryo transfer :  multifoetal pregnancy reduction versus expectant management

2019 ◽  
Author(s):  
Pei Cai ◽  
Yan Ouyang ◽  
Fei Gong ◽  
Xihong Li
Keyword(s):  
Preterm Birth ◽  
Pregnancy Outcomes ◽  
Expectant Management ◽  
Triplet Pregnancy ◽  
Vitro Fertilization ◽  
Group A ◽  
Group B ◽  
Singleton Pregnancies ◽  
Ivf Et

Abstract Background: Trichorionic triplet pregnancy reduction to twin pregnancy is associated with a lower risk of preterm delivery but not with a lower risk of miscarriage. However, data on dichorionic triamniotic (DCTA) triplet pregnancy outcomes are lacking. This study aimed to compare the pregnancy outcomes of DCTA triplets conceived via in vitro fertilization-embryo transfer (IVF-ET) managed expectantly or reduced to a monochorionic (MC) singleton or monochorionic diamniotic (MCDA) twins at 11-13 +6 gestational weeks. Method s : Two hundred ninety-eight patients with DCTA triplets conceived via IVF-ET between 2012 and 2016 were retrospectively analysed. DCTA triplets with three live foetuses were reduced to a MC singleton (group A) or MCDA twins (group B) or underwent expectant management (group C). Each multifoetal pregnancy reduction (MFPR) was performed at 11-13 +6 gestational weeks. Pregnancy outcomes in the 3 groups were compared. Results: Eighty-four DCTA pregnancies were reduced to MC singleton pregnancies, 149 were reduced to MCDA pregnancies, and 65 were managed expectantly. There were no significant differences among groups A, B, and C in miscarriage rate (8.3 vs. 7.4 vs. 10.8%, respectively) and live birth rate (90.5 vs. 85.2 vs. 83.1%, respectively) (P > 0.05). Group A had significantly lower rates of preterm birth (8.3 vs. 84.6%; odds ratio (OR) 0.017, 95% confidence interval (CI) 0.006-0.046) and low birth weight (LBW; 9.2 vs. 93.2%; OR 0.007, 95% CI 0.003-0.020) than group C (P < 0.001). Group B had significantly lower preterm birth (47.0 vs. 84.6%; OR 0.161, 95% CI 0.076-0.340) and LBW rates (58.7 vs. 93.2%; OR 0.103, 95% CI 0.053-0.200) than group C (P < 0.001). Group A had significantly lower preterm birth (8.3 vs. 47.0%; OR 0.103, 95% CI 0.044-0.237; P < 0.001), LBW (9.2 vs. 58.7%; OR 0.071, 95% CI 0.032-0.162; P < 0.001) and perinatal death rates (1.3 vs. 9.1%; OR 0.132, 95% CI 0.018-0.991; P = 0.021) than group B. Conclusion: The MFPR of DCTA triplets to singleton or MCDA pregnancies was associated with better pregnancy outcomes compared to expectant management. DCTA triplets reduced to singleton pregnancies had better perinatal outcomes than DCTA triplets reduced to MCDA pregnancies.

scholarly journals Pregnancy outcomes of dichorionic triamniotic triplet pregnancies after in vitro fertilization-embryo transfer : multifoetal pregnancy reduction versus expectant management

2020 ◽  
Author(s):  
Pei Cai ◽  
Yan Ouyang ◽  
Fei Gong ◽  
Xihong Li
Keyword(s):  
Preterm Birth ◽  
Pregnancy Outcomes ◽  
Expectant Management ◽  
Triplet Pregnancy ◽  
Vitro Fertilization ◽  
Group A ◽  
Group B ◽  
Singleton Pregnancies ◽  
Ivf Et

Abstract Background: Trichorionic triplet pregnancy reduction to twin pregnancy is associated with a lower risk of preterm delivery but not with a lower risk of miscarriage. However, data on dichorionic triamniotic (DCTA) triplet pregnancy outcomes are lacking. This study aimed to compare the pregnancy outcomes of DCTA triplets conceived via in vitro fertilization-embryo transfer (IVF-ET) managed expectantly or reduced to a monochorionic (MC) singleton or monochorionic diamniotic (MCDA) twins at 11-13 +6 gestational weeks. Method s : Two hundred ninety-eight patients with DCTA triplets conceived via IVF-ET between 2012 and 2016 were retrospectively analysed. DCTA triplets with three live foetuses were reduced to a MC singleton (group A) or MCDA twins (group B) or underwent expectant management (group C). Each multifoetal pregnancy reduction (MFPR) was performed at 11-13 +6 gestational weeks. Pregnancy outcomes in the 3 groups were compared. Results: Eighty-four DCTA pregnancies were reduced to MC singleton pregnancies, 149 were reduced to MCDA pregnancies, and 65 were managed expectantly. There were no significant differences among groups A, B, and C in miscarriage rate (8.3 vs. 7.4 vs. 10.8%, respectively) and live birth rate (90.5 vs. 85.2 vs. 83.1%, respectively) (P > 0.05). Group A had significantly lower rates of preterm birth (8.3 vs. 84.6%; odds ratio (OR) 0.017, 95% confidence interval (CI) 0.006-0.046) and low birth weight (LBW; 9.2 vs. 93.2%; OR 0.007, 95% CI 0.003-0.020) than group C (P < 0.001). Group B had significantly lower preterm birth (47.0 vs. 84.6%; OR 0.161, 95% CI 0.076-0.340) and LBW rates (58.7 vs. 93.2%; OR 0.103, 95% CI 0.053-0.200) than group C (P < 0.001). Group A had significantly lower preterm birth (8.3 vs. 47.0%; OR 0.103, 95% CI 0.044-0.237; P < 0.001), LBW (9.2 vs. 58.7%; OR 0.071, 95% CI 0.032-0.162; P < 0.001) and perinatal death rates (1.3 vs. 9.1%; OR 0.132, 95% CI 0.018-0.991; P = 0.021) than group B. Conclusion: The MFPR of DCTA triplets to singleton or MCDA pregnancies was associated with better pregnancy outcomes compared to expectant management. DCTA triplets reduced to singleton pregnancies had better perinatal outcomes than DCTA triplets reduced to MCDA pregnancies.

scholarly journals Single-unit transfusions of RBC enzymatically converted from group B to group O to A and O normal volunteers

Blood ◽  
1991 ◽  
Vol 77 (6) ◽  
pp. 1383-1388 ◽  
Author(s):  
LL Lenny ◽  
R Hurst ◽  
J Goldstein ◽  
LJ Benjamin ◽  
RL Jones
Keyword(s):  
Single Unit ◽  
Small Volume ◽  
Survival Times ◽  
Chronic Anemia ◽  
Group A ◽  
Alpha Galactosidase ◽  
Group B ◽  
Sustained Increase

Abstract Full-unit transfusions of RBC enzymatically converted from group B to group O by treatment with alpha-galactosidase (ECO RBC) to group O and A normal healthy individuals exhibit excellent in vivo survival times (24-hour survival 95.1% +/- 2.3%, T50 36.9 +/- 4.6 days). These results confirm our earlier findings describing ECO RBC in vitro viability and normal in vivo survival time after small-volume infusions. No significant increase in pretransfusion anti-B titer or score is observed in either group O or A subjects provided that sufficient enzyme is used to treat the cells: Cells transfused to group O recipients require higher levels of enzyme (185 to 200 U/mL RBC) than those infused to group A (90 U/mL RBC). Two separate single-unit transfusions of ECO RBC to one group O recipient (4.5 months apart) also survived normally (24-hour survival 96% and 92%, T50 40 and 36 days) and did not increase preexisting anti-B levels in this subject. ECO RBC were not agglutinated or lysed by recipient sera before or after transfusion. Similarly, no antibody development to the alpha- galactosidase used in cell treatment (and washed from the product before transfusion) could be detected in any subject. The sustained increase in hemoglobin levels after transfusion of ECO RBC suggests that this product will be useful in treatment of acute and chronic anemia.

114 CRYOTOP VITRIFICATION FOR IN VITRO-PRODUCED BUFFALO (BUBALUS BUBALIS) EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 174 ◽  
Author(s):  
A. De Rosa ◽  
L. Attanasio ◽  
L. Boccia ◽  
G. Pellerano ◽  
G. Campanile ◽  
...  
Keyword(s):  
Survival Rate ◽  
Liquid Nitrogen ◽  
Sucrose Solution ◽  
Bubalus Bubalis ◽  
Thin Strip ◽  
Embryo Survival ◽  
Vitro Fertilization ◽  
Group A ◽  
Group B

The aim of this study was to compare the efficiency of different combinations of cryoprotectants for vitrification of IVP buffalo (Bubalus bubalis) embryos by the cryotop method (Kuwayama et al. 2005 RBM Online 11, 300–308). In group A, we evaluated the vitrification and warming solutions previously used to vitrify buffalo embryos in French straws (Gasparrini et al. 2001 Theriogenology 55, 307). Embryos were equilibrated in 1.4 M glycerol for 5 min and in 1.4 M glycerol and 3.6 M ethylene glycol (EG) for an additional 5 min. After being transferred into 3.4 M glycerol and 4.6 M EG for 25 s, individual embryos were picked up in an extremely small volume (&lt;0.1 �L) of vitrification solution and placed on the top of a very fine polypropylene strip (0.4 mm wide � 20 mm long � 0.1 mm thick) attached to a hard plastic handle, kindly provided by M. Kuwayama. Each embryo was placed onto the thin strip of the Cryotop and immediately submerged into liquid nitrogen. For warming, the strip of the Cryotop was immersed directly into a 0.5 M sucrose solution; embryos were retrieved and transferred into 0.25 M sucrose for 5 min before culture in SOF medium. In group B, we examined the vitrification and warming solutions previously used for OPS vitrification of buffalo embryos (De Rosa et al. 2006 Reprod. Fertil. Dev. 18, 153). Embryos were equilibrated in 7.5% EG + 7.5% dimethyl sulfoxide (DMSO) for 3 min before transfer into 16.5% EG + 16.5% DMSO + 0.5 M sucrose. After 25 s, they were placed on the cryotop, as previously described, and submerged into liquid nitrogen. For warming, embryos were recovered into a 0.25 M sucrose solution for 1 min, transferred into 0.15 M sucrose for 5 min, and cultured in SOF. IVP buffalo embryos of excellent quality that, by Day 7 of culture (Day 0 = in vitro fertilization), had reached the blastocyst stage (n = 44 and 53 for groups A and B, respectively), over 6 replicates, were vitrified. Embryo survival rate was determined as the percentage of vitrified-warmed embryos undergoing further development during a 24-h in vitro culture period. Differences between methods were analyzed by chi-square test. A significantly higher embryo survival rate was recorded in Group B compared to Group A (67.9 vs. 43.2% respectively; P &lt; 0.05). In conclusion, it was demonstrated that cryotop vitrification, with the combination of cryoprotectants used in group B, is a valid tool to cryopreserve IVP buffalo blastocysts.

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