scholarly journals Production of viable bovine embryos by intracytoplasmic sperm injection of oocytes harvested from slaughtered old cows

2021 ◽  
Vol 26 (1) ◽  
pp. 7-14
Author(s):  
Mihai Cenariu ◽  
Mihai Borzan ◽  
Sorin Dan ◽  
Remus Chiorean ◽  
Emoke Pall
Keyword(s):  
In Vitro Fertilization ◽  
Intracytoplasmic Sperm Injection ◽  
Fertilization Rate ◽  
Fertilization Success ◽  
Vitro Fertilization ◽  
Bull Semen ◽  
Intracytoplasmic Injection ◽  
Lower Fertility

Abstract: (1) Background: Intracytoplasmic sperm injection (ICSI) is currently used to increase fertilization success by avoiding several oocyte or sperm deficiencies that would normally prevent conception after in vivo fertilization or classical in vitro fertilization. This paper aimed at improving the in vitro fertilization protocol of bovine oocytes, harvested from old cows after slaughtering, using intracytoplasmic sperm injection; (2) Methods: Oocytes were harvested by puncture of follicles from ovaries obtained from slaughtered old cows, followed by aspiration. Out of the 127 cumulus-oocyte complexes that were harvested, 84 (66.14%) were declared suitable for cultivation, after morphological evaluation. Following oocyte maturation for 22 hours, 77 cumulus-oocyte complexes were morphologically intact and could undergo the steps required for intracytoplasmic injection of spermatozoa. Frozen-thawed bull semen was used for ICSI and the 77 fertilized oocytes were kept for 24 hours in an atmosphere enriched with 5% CO2.; (3) Results: Fertilized oocytes transformed into 46 zygotes (fertilization rate of 59.74%), while after 168 h of cultivation 38 transferable compact morulae or early blastocysts were obtained; (4) Conclusions: Intracytoplasmic sperm injection can represent a viable alternative to classical IVF, when oocytes or sperm with lower fertility are used.

Development and implementation of intracytoplasmic sperm injection (ICSI)

1995 ◽  
Vol 7 (2) ◽  
pp. 211 ◽  
Author(s):  
GD Palermo ◽  
J Cohen ◽  
M Alikani ◽  
A Adler ◽  
Z Rosenwaks
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Human Sperm ◽  
Fertilization Rate ◽  
Semen Parameters ◽  
Ongoing Pregnancy Rate ◽  
Vitro Fertilization ◽  
Human Oocytes ◽  
Intracytoplasmic Injection ◽  
Previously Treated

The purpose of this paper is to elucidate the experimental steps that led to the development of intracytoplasmic sperm injection (ICSI) and its application in the human. ICSI has become the most successful micromanipulation procedure for treating male infertility. A total of 355 in vitro fertilization (IVF) cycles utilizing ICSI are described; 180 couples were previously treated in 509 IVF cycles but achieved no fertilization and 175 couples could not be treated by IVF because of extremely poor semen parameters. Of the 3063 metaphase II (M II) oocytes retrieved, 2970 were injected with a survival rate of 93.6%, yielding 1917 bipronuclear zygotes (64.5%). In 148 patients, a foetal heart was evidenced by ultrasound; 11 of these patients miscarried between 7 and 13 weeks of gestation. The ongoing pregnancy rate was 38.6% (137/355) per retrieval and 40.5% (137/338) per embryo replacement. At the time of writing, there were 22 deliveries and one therapeutic abortion for a trisomy 21 chromosomal abnormality. In addition, 66 singleton, 37 twin, 10 triplet and 1 quadruplet pregnancies were ongoing. The concentration of motile spermatozoa in the ejaculate only slightly influenced the fertilization rate (P < 0.001) and the pregnancy outcome (P < 0.01). A preliminary injection procedure utilizing intracytoplasmic injection of isolated sperm heads was performed in 35 M II human oocytes with resultant fertilization and cleavage rates of 74% and 73% respectively. Skills in ICSI were acquired by injecting hamster and unfertilized human oocytes with human sperm. ICSI can be used to successfully treat couples who have failed IVF or who have too few spermatozoa for conventional in vitro insemination.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Effect of sperm DNA fragmentation on clinical outcomes for Chinese couples undergoing in vitro fertilization or intracytoplasmic sperm injection

2016 ◽  
Vol 44 (6) ◽  
pp. 1283-1291 ◽  
Author(s):  
Lin-Tao Xue ◽  
Rui-Xue Wang ◽  
Bing He ◽  
Wei-Ying Mo ◽  
Li Huang ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Dna Fragmentation ◽  
Intracytoplasmic Sperm Injection ◽  
Fertilization Rate ◽  
Sperm Dna Fragmentation ◽  
Roc Curve Analysis ◽  
Vitro Fertilization ◽  
Fragmentation Rate ◽  
Sperm Dna

Objective To investigate the effect of sperm DNA fragmentation on the fertilization rate, embryo development and pregnancy outcome of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in a cohort of Chinese couples. Methods Infertile couples that had undergone assisted reproductive technology at our centre between January 2011 and December 2013 were included in this retrospective study. Fractions of prepared sperm samples were evaluated for sperm DNA fragmentation on the day of oocyte recovery. Results Of the 550 couples selected, 415 had undergone IVF and 135 ICSI. Sperm DNA fragmentation rate was significantly negatively correlated with the fertilization rate in the ICSI cycles but not the IVF cycles. No association was found between sperm DNA fragmentation and cleavage rate or good quality embryo formation rates in IVF or ICSI cycles. Receiver operating characteristic (ROC) curve analysis showed that the sperm DNA fragmentation rate was a statistically significant prognostic indicator of the clinical fertilization rate in ICSI cycles; a rate > 22.3% was associated with a lower fertilization rate following ICSI compared with a rate ≤ 22.3%. Conclusions High values of sperm DNA fragmentation were associated with a low fertilization rate following ICSI but were not associated with alterations in pregnancy or live birth rates in either ICSI or IVF in this cohort of Chinese couples.

scholarly journals Hubungan Anti-Mullerian Hormone (AMH) pada Long dan Short Protocol Terhadap Fertilization Rate Pasien In Vitro Fertilization (IVF)

2021 ◽  
Vol 4 (2) ◽  
pp. 151-161
Author(s):  
Rizki Amalia Wahid ◽  
◽  
Edwin Armawan ◽  
ono Djuwantono
Keyword(s):  
In Vitro Fertilization ◽  
Intracytoplasmic Sperm Injection ◽  
Fertilization Rate ◽  
Chi Square ◽  
Vitro Fertilization ◽  
Fertility Clinic ◽  
Long Protocol

Abstrak Tujuan: Untuk mengevaluasi pengaruh kadar anti-mullerian hormone (AMH) dengan fertilization rate (FR) dan menilai perbedaan pengaruh jenis protokol (long protocol (LP) dan short protocol (SP)) pada tiap tingkat cadangan ovarium terhadap FR pada pasien in vitro fertilization (IVF) dengan Intracytoplasmic Sperm Injection. Metode: Data sekunder dari rekam medis pasien yang menjalani IVF di Aster Fertility Clinic Rumah Sakit Umum Pendidikan dr. Hasan Sadikin pada tahun 2016-2020 dan Bandung Fertility Centre Rumah Sakit Ibu Anak Limijati pada tahun 2018-2019. Penelitian ini analitik observational dengan metode Cohort retrospektif. Hubungan antara dua data kategorik diuji dengan uji chi-square dan uji Kruskal-Wallis digunakan pada data numerik dengan distribusi yang tidak rata pada lebih dari 2 kelompok, Hasil: Hasil data diperoleh nilai rerata kadar AMH secara keseluruhan adalah 3.30 ng/ml dengan rerata capaian FR sebesar 71.97%. Berdasarkan metode IVF yang dipilih, mayoritas pasien menjalani pengobatan SP 54.4% (rerata FR 72.80%) dibandingkan dengan LP 45.6% (rerata FR 70.97%). Tidak ditemukan hubungan yang bermakna antara kadar AMH dengan FR, dinyatakan dengan nilai p=0.977. Kadar AMH terhadap FR bila dipisahkan menurut protokol terapi yang diberikan tidak menunjukkan perbedaan yang bermakna pada masing-masing protokol (LP p=0,763; SP p=0,843). Mengenai hubungan antara protokol IVF dengan FR juga tidak diperoleh perbedaan yang signifikan secara statistik dengan nilai p=0,27 (RR 1.17 (0.62-2.15); CI 95%). Penggobatan menggunakan LP (p=0,770) maupun SP (p=0.845) tidak memberikan pengaruh yang bermakna terhadap FR pada setiap kategori AMH. Kesimpulan: Tidak ada pengaruh kadar AMH dan protokol terapi terhadap FR. Kata kunci : In Vitro Fertilization, Fertilization Rate, Anti-Mullerian Hormone, Protokol Stimulasi Ovarium

312 EXPOSURE OF SPERMATOZOA TO SOLUBILIZED EXTRACTS OF THE OVIDUCTAL EPITHELIUM APICAL PLASMA MEMBRANE ENHANCES FERTILIZATION IN PORCINE IN VITRO FERTILIZATION

2007 ◽  
Vol 19 (1) ◽  
pp. 272
Author(s):  
N. Satake ◽  
A. K. Alhaider ◽  
W. V. Holt ◽  
P. F. Watson
Keyword(s):  
Plasma Membrane ◽  
In Vitro Fertilization ◽  
Plasma Membranes ◽  
Fertilization Rate ◽  
Apical Plasma Membrane ◽  
In Vitro Production ◽  
Vitro Fertilization ◽  
Fertilization Rates

In vitro production (IVP) of porcine embryos is currently suboptimal compared with IVP in species such as mice and cattle. In vitro fertilization (IVF) usually involves the co-culture of oocytes and spermatozoa in a medium droplet. Oocyte quality is the focus of many studies. In vivo, the quality of spermatozoa is as important as the oocyte, and females have many mechanisms to select the highest quality spermatozoa for their oocytes. Oviductal proteins have been shown to affect sperm motility of subpopulations within an ejaculate. The present study was carried out to investigate normal and polyspermic fertilization rates of spermatozoa exposed to oviductal epithelial apical plasma membrane (APM) proteins, a mixture of peripheral proteins extracted by 1 M NaCl from isolated oviductal apical plasma membranes, prior to co-culture with oocytes in IVF. Porcine oocytes were aspirated from ovaries and grade I quality oocytes (cumulus–oocyte complexes with a spherical shape, visible nucleus, even-density cytoplasm, and multiple layers of cumulus cells) were selected and matured for 48 h in TCM-199 supplemented with LH (0.5 �g mL-1), FSH (0.5 �g mL-1), and EGF (10 ng mL-1). Ejaculates were washed through a Percoll gradient to obtain a concentrated pellet. Spermatozoa were diluted in capacitation–fertilization medium in the presence or absence of APM proteins (100 �g mL-1), incubated for 10 min, and then co-cultured with oocytes for 6 h in modified Tween medium B with milk powder medium (Abeydeera and Day 1997 Theriogenology 48, 537–544) supplemented with BSA (0.4%) and sodium bicarbonate (5 mM). Presumptive zygotes were cultured in NCSU23 medium for a further 48 h. The oocytes/zygotes were then fixed and stained with propidium iodide for evaluation by confocal microscopy for fertilization and cleavage (n = 1235 oocytes). Fertilization rates were compared between treatments in a chi-squared test using the Mantel-Haenszel approach. The overall fertilization rate was significantly higher (78 vs. 86%) when spermatozoa were incubated in the presence of APM proteins (P &lt; 0.05), and in the group of fertilized oocytes, polyspermic fertilization (47 vs. 21%) was significantly reduced when spermatozoa were exposed to APM proteins (P &lt; 0.01). However, cleavage rates were not different. These results suggest that exposure of spermatozoa to APM proteins prior to IVF increases the fertilization rate and decreases the incidence of polyspermic penetration.

Comparison of the Outcome of Conventional in vitro Fertilization and Intracytoplasmic Sperm Injection in Moderate Male Infertility from Ejaculate

2013 ◽  
Vol 94 (1) ◽  
pp. 111-116 ◽  
Author(s):  
Bao-Guo Xie ◽  
Yuan-Hua Huang ◽  
Wei-Jie Zhu ◽  
Song Jin
Keyword(s):  
In Vitro Fertilization ◽  
Male Infertility ◽  
Intracytoplasmic Sperm Injection ◽  
Semen Analysis ◽  
Implantation Rate ◽  
Fertilization Rate ◽  
Vitro Fertilization ◽  
Good Quality Embryo ◽  
Implantation And Pregnancy Rates

Objective: To evaluate whether couples with moderate male infertility should be treated with conventional in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Patients and Methods: A total of 249 couples with moderate male infertility undergoing their first IVF/ICSI cycle were enrolled in the study. The couples were divided into two groups according to the results of semen analysis: moderate oligozoospermia (O group) and moderate oligoasthenozoospermia (OA group). Sibling oocytes were randomized into groups to be inseminated either by conventional IVF or ICSI. Fertilization rate, embryo quality, implantation rate, and clinical pregnancy rate were examined. Results: There was no difference in the fertilization, implantation, and pregnancy rates between conventional IVF and ICSI in either the O group or OA group (p > 0.05). Additionally, in the OA group, the good quality embryo rate was similar after IVF or ICSI (p > 0.05). However, in the O group, the good quality embryo rate was significantly higher after ICSI than after IVF (p < 0.05). Conclusions: Couples with moderate oligozoospermia or moderate oligoasthenozoospermia did not influence the major indices of IVF. Because of the uncertainties concerning the safety of ICSI, couples with moderate oligozoospermia or moderate oligoasthenozoospermia need not be subjected to this procedure.

scholarly journals Clinical Outcomes of Rescue Intracytoplasmic Sperm Injection at Different Timings Following In Vitro Fertilization

2021 ◽  
Author(s):  
Yuki Shiraiwa ◽  
Noritoshi Enatsu ◽  
Kazuki Yamagami ◽  
Koyu Furuhashi ◽  
Toshiroh Iwasaki ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Intracytoplasmic Sperm Injection ◽  
Polar Body ◽  
Fertilization Rate ◽  
Optimal Timing ◽  
Vitro Fertilization ◽  
Polar Body Analysis ◽  
Second Polar Body ◽  
Rescue Icsi

Background: Although rescue intracytoplasmic sperm injection (r-ICSI) is extensively used worldwide, the indication of r-ICSI and its optimal timing remains obscure. This study aimed to assess the outcomes of r-ICSI following in vitro fertilization in different timings when fertilization is confirmed. Methods: This study included 5,156 cycles (47,785 eggs). Fertilization was confirmed by polar body analysis after 4 and 6 hr of coincubation of the sperm and oocyte. Oocytes that underwent IVF were divided into two groups based on the time when a second polar body was detected in more than 30% of all oocytes (Four-hr group and six-hr group). If the second polar body was not detected or was present in less than 30% of all oocytes after six hr of coincubation, rescue-ICSI (r-ICSI) was performed for oocytes without a second polar body (r-ICSI group). Results: The fertilization rates of two pronuclear (2PN) oocytes in the three groups (Four-hr group, six-hr group, and r-ICSI group) were 70.7%, 51.3%, and 58.0%, respectively. The blastocyst formation rates were 62.8%, 53.4%, and 42.9%, respectively. Conclusion: Performing r-ICSI after six hr of coincubation can salvage cases with fertilization failure in IVF. The higher fertilization rate of r-ICSI indicates that all oocytes without signs of fertilization after six hr of coincubation should undergo r-ICSI.

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