Pregnancy and calving rates following transfer of in-vitro-produced river and F1 (river × swamp) buffalo (Bubalus bubalis) embryos in recipients on natural oestrus or synchronised for ovulation

2007 ◽  
Vol 19 (5) ◽  
pp. 670 ◽  
Author(s):  
Xianwei Liang ◽  
Xiufang Zhang ◽  
Bingzhuang Yang ◽  
Mingtang Cheng ◽  
Fenxiang Huang ◽  
...  
Keyword(s):  
Bubalus Bubalis ◽  
Sperm Cells ◽  
In Vitro Production ◽  
In Vitro Development ◽  
Antral Follicles ◽  
Swamp Buffaloes ◽  
Swamp Buffalo ◽  
Vitro Development ◽  
Vitro Production

The main objective of this study was to compare pregnancy and calving rates following transfer of in-vitro-produced fresh river and F1 (river × swamp) buffalo embryos in recipients synchronised by Ovsynch protocol or following natural oestrus. River embryos were produced from cumulus–oocyte complexes (COCs) derived by ovum pick up (OPU) on 40 Murrah and Nili-Ravi donor buffaloes over a twice-weekly collection schedule for 120 single OPUs. F1 embryos were produced by fertilisation of swamp COCs recovered from abattoir ovaries coincubated with river sperm cells. Both groups of embryos were produced following the same protocol for in vitro production. With regard to the OPU source of COCs, 923 antral follicles were punctured and 647 COCs were recovered (70%). From 462 selected COCs for IVM, 257 (55.6%) cleaved zygotes were recorded leading to 93 blastocysts (20.1%). In total, 590 swamp COCs were aspirated from abattoir ovaries and 476 were selected for IVM leading to 270 (56.7%) cleaved zygotes and resulting in 137 blastocysts (28.8%). River and F1 embryos were transferred between Day 6 to 7 of in vitro development, corresponding to blastocyst–expanding blastocyst, into F1 recipients synchronised by Ovsynch and swamp buffaloes following natural oestrus, respectively, each of them receiving two embryos. According to palpation per rectum of the ovaries at the time of embryo transfer, 26 of the 47 (55.3%) F1 recipients synchronised by Ovsynch were considered suitable for transfer, resulting in seven pregnancies (26.9%) and four calvings (15.3%) owing to three abortions occurring between 2 and 3 months of pregnancy. In total, 29 swamp recipients following natural oestrus were judged suitable as recipients, resulting in 12 pregnancies (41.4%) and 10 calvings (34.5%) owing to two abortions at 2 and 3 months of gestation respectively. Pregnancy and calving rates following transfer of river and F1 embryos were similar. Likewise, weight at birth of calves derived from transfer of river and F1 embryos was not different: 30.5 ± 1.4 and 32.9 ± 2.4 respectively. Pregnancy and calving rates following AI in a group of river and swamp buffaloes considered for reference in this study were similar to recipients carrying in-vitro-produced embryos. Collectively, no apparent postnatal complications were recorded in resulting live calves.

scholarly journals In vitro development of sugarcane seedlings using ethephon or gibberellin

2018 ◽  
Vol 8 (2) ◽  
pp. 389-395
Author(s):  
Luciane De Siqueira Mendes ◽  
Marcia Eugenia Amaral Carvalho ◽  
Willian Rodrigues Macedo ◽  
Paulo Roberto de Camargo e Castro
Keyword(s):  
Gibberellic Acid ◽  
Dry Mass ◽  
In Vitro Production ◽  
In Vitro Development ◽  
Reduced Height ◽  
Potential Strategy ◽  
Vitro Development ◽  
Vitro Production ◽  
Different Doses

The use of plant growth regulators is directly related to the success of in vitro propagation, which is an advantageous alternative to obtain seedlings on a commercial scale. This study aimed to evaluate the in vitro development of ‘IAC 95-5000’ sugarcane seedlings after the addition of different doses of ethephon (0, 25, 50, 100 and 200 mg L-1) or gibberellic acid (0, 2.5, 5.0, 7.5 and 10.0 mg L-1) to the culture medium. Ethephon increased the number of tillers (up to 231.70%), reduced height of the main tiller (44.66 to 60.47%), and did not affect the shoot´s fresh and dry mass. On the other hand, gibberellin decreased the number of tillers and negatively changed biomass partitioning. It is concluded that the use of ethephon is a potential strategy to enhance in vitro production of ‘IAC 95-5000’ sugarcane seedlings, since it increased the number of usable shoots in subsequent subcultures, and its effects on height reduction can be reversible. However, the use of the tested doses of gibberellic acid is not recommended, because it impaired seedling development of this sugarcane variety.

scholarly journals α-Tocopherol and l-ascorbic acid increase the in vitro development of IVM/IVF swamp buffalo (Bubalus bubalis) embryos

animal ◽  
2008 ◽  
Vol 2 (10) ◽  
pp. 1486-1490 ◽  
Author(s):  
K. Saikhun ◽  
T. Faisaikarm ◽  
Z. Ming ◽  
K.H. Lu ◽  
Y. Kitiyanant
Keyword(s):  
Ascorbic Acid ◽  
Bubalus Bubalis ◽  
In Vitro Development ◽  
Swamp Buffalo ◽  
Vitro Development

Roscovitine Treatment Improves Synchronization of Donor Cell Cycle in G0/G1 Stage and In Vitro Development of Handmade Cloned Buffalo (Bubalus bubalis) Embryos

2012 ◽  
Vol 14 (2) ◽  
pp. 146-154 ◽  
Author(s):  
Naresh L. Selokar ◽  
Monika Saini ◽  
Mushariffa Muzaffer ◽  
G. Krishnakanth ◽  
Ambika P. Saha ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Donor Cell ◽  
Bubalus Bubalis ◽  
In Vitro Development ◽  
Vitro Development

298 SPERM CAPACITATED WITH CALCIUM IONOPHORE AS A VECTOR FOR IN VITRO PRODUCTION OF BOVINE TRANSGENIC EMBRYOS

2006 ◽  
Vol 18 (2) ◽  
pp. 256
Author(s):  
R. Simões ◽  
M. P. Milazzotto ◽  
C. Yamada ◽  
W. B. Feitosa ◽  
A. R. S. Coutinho ◽  
...  
Keyword(s):  
Cell Monolayer ◽  
Calcium Ionophore ◽  
Control Group ◽  
Sperm Cells ◽  
In Vitro Production ◽  
Successful Technique ◽  
Blastocyst Rate ◽  
Transgenic Embryos ◽  
Vitro Production

Production of transgenic mouse embryos by microinjection is a well established and successful technique. However, when microinjection protocols were used for bovine, the amount of the oocyte lipid content did not allow the production of bovine transgenic embryos. Sperm-mediated gene transfer (SMGT) is an alternative for this species because it has lower cost and does not require microinjection handling. One of the procedures to introduce exogen DNA into oocytes is by means of sperm capacitated with calcium ionophore (CaI). The aim of this work was to evaluate different CaI concentrations ([CaI]), sperm incubation times with CaI (tCa), and incubation times of sperm capacitated with DNA (tDNA) (EYFP; Clontech, Palo Alta, CA, USA) to establish a satisfactory method for IVP of bovine transgenic embryos. Slaughterhouse oocytes with compact cumulus and uniform ooplasm were in vitro maturated in TCM-199 medium + 10% FCS + FSH + hCG + estradiol (E2) + piruvate + gentamicin under 5% CO2 in air, at 39�C and high humidified atmosphere for 24 h. Semen was thawed in a water bath at 37�C for 30 s and separated by Percoll gradient (45/90%) at 600g for 30 min. After this procedure, sperm cells were washed in TALP-semen medium by centrifugation at 200g for 5 min at room temperature. Supernatant was removed and capacitation (5 � 106 spermatozoa/group) was induced with CaI (250 nM or 500 nM for 1 or 5 min). Capacitated sperm cells were incubated with 500 ng/mL DNA for 1 or 2 h. Nontreated spermatozoa were used as control group. Sperm cells (1 � 105) were used to inseminate 20 oocytes/90 mL microdroplets for 18 h. The presumptive zygotes were co-cultured in SOFaa medium with a granulosa cell monolayer under high humidified atmosphere, at 39�C and 5% CO2 in air. Blastocyst rates were analyzed by ANOVA. Independent variables were replicate, [CaI], tCa, tDNA, and the double and triple interactions among the last three variables; when appropriate, means were compared by orthogonal contrasts. There was [CaI] � tCa � tDNA interaction for blastocyst rate (P < 0.02). Treatments with 250 nM ([CaI]), 5 min (tCaI), and 1 h (tDNA) or 500 nM ([CaI]), 1 min (tCaI), and 1 h (tDNA) resulted in 36.1% and 37.4% blastocyst rates, respectively, similar to the control group (30.5%; P > 0.4). These results demonstrated that it is possible to capacitate spermatozoa with CaI to produce transgenic embryos, without alteration of blastocyst rate. This work was supported by FAPESP 03/08542-5 and 03/07456-8.

292 EXPRESSION PROFILES OF STRESS AND METABOLIC MARKER GENES DURING IN VITRO PRODUCTION OF BUFFALO (BUBALUS BUBALIS) EMBRYOS

2006 ◽  
Vol 18 (2) ◽  
pp. 253
Author(s):  
R. Rajhans ◽  
G. S. Kumar ◽  
G. T. Sharma
Keyword(s):  
Bubalus Bubalis ◽  
Marker Genes ◽  
Hsp 70 ◽  
In Vitro Production ◽  
Glucose Transporter 1 ◽  
Total Rna ◽  
Gene Transcripts ◽  
Vitro Production ◽  
Glut1 Gene

An increased understanding of the pre-implantation embryo developmental stage, with respect to physiological interaction of embryo with its micromilieu both in vivo and in vitro, is imperative to comprehend the events of pre-implantation development.The objective of the present study was to examine the temporal expression of heat shock protein 70 (Hsp-70) and glucose transporter 1 (Glut1) genes in pre-implantation-stage buffalo embryos produced under the standard in vitro production (IVP) system. Embryos were produced from slaughterhouse ovaries employing standard in vitro embryo production protocol, and presumptive zygotes produced following IVM/IVF were cultured in vitro in mSOF under mineral oil; FCS (10%) was added at 48 h post-insemination (hpi).The time series of development at stages being zygote (18-20 hpi), 2-1 cell (48 hpi), 8-16 cell (94-96 hpi), morula (120-144 hpi), and blastocyst (168-192 hpi), pre-implantation embryos conforming to the above developmental pattern were considered as 'fast-cleaving embryos', and all the embryos that did not conform to the above developmental timing were regarded as 'slow-cleaving embryos'. Pools of immature oocytes (IM, 120), Matured oocytes (MO, 120), 8-16 cell stages (8-16, 70), morula (M, 28), and blastocyst (B, 9) were collected and prepared for total RNA isolation and RT-PCR for the specific transcripts, with �-actin as loading control. The total RNA content ranged from 2.5 to 5.0 ng per oocyte/embryo. Presence of Hsp 70 and Glut1 gene transcripts was assessed in different stages of buffalo pre-implantation embryos using primers designed from bovine Hsp 70 and Glut1 by using the OLIGO program. RT was standardized using the embryo equivalent of 1-10 oocytes/embryo as the template as described by Arcellana-Panlilio and Schultz 1993 (Methods Enzymol. 225, 303-328) with PCR conditions being 59�C and 62�C for 45 s with 39 cycles for Glut1 and Hsp 70 gene transcripts, respectively. Amplicons were subjected to restriction digestion and sequencing (Acc. No. AJ812563, AJ812564). The expression of Hsp 70 throughout pre-implantation development in the fast-cleaving embryos indicated their maternal and zygotic origin, but transcripts of the Hsp 70 gene, represented by a single 488-bp amplicon, were not detected in slow-cleaving embryos, suggesting altered zygotic expression. Glut1 expression was prominent from the 8-16 cell stage, indicating a metabolic shift from pyruvate to glucose after the pre-compaction stage. For slow-cleaving embryos, transcripts of the Glut1 gene, represented by a single 327-bp amplicon, were absent during morula- and blastocyst-stage embryos, indicating the poor developmental competence of these embryos, which morphologically appeared normal. These transcription patterns reflect the embryonic response to the in vitro culture conditions and also correlate with the embryo quality and the speed of development of the pre-implantation buffalo (Bubalus bubalis) embryos.

scholarly journals 257 EFFECTS OF GLUTAMINE AND HYPOTAURINE ON OXIDATIVE STRESS OF PORCINE EMBRYOS CULTURED IN VITRO

2005 ◽  
Vol 17 (2) ◽  
pp. 278 ◽  
Author(s):  
C. Suzuki ◽  
K. Yoshioka
Keyword(s):  
Oxidative Stress ◽  
Subsequent Development ◽  
In Vitro Production ◽  
Vitro Fertilization ◽  
H 41 ◽  
H2o2 Level ◽  
Vitro Development ◽  
Vitro Production ◽  
Number Of Cells

We previously developed an in vitro production system for porcine embryos and reported that the addition of glutamine and hypotaurine during in vitro culture improved blastocyst yield and the total number of cells in the blastocysts. Glutamine and hypotaurine might reduce oxidative stress, allowing the development of embryos cultured in vitro, because glutamine reportedly protects embryos against oxidative stress by helping to maintain intracellular levels of cysteine, a precursor of glutathione (GSH), and hypotaurine is a potent antioxidant. In the present study we evaluated the effects of the presence of glutamine and hypotaurine from Day 2 (Day 0 = the day of in vitro fertilization) to Day 3 on oxidative stress during in vitro development of porcine embryos. Porcine cumulus-oocytes complexes from prepubertal gilts were matured and fertilized in vitro using frozen-thawed ejaculated boar semen (Yoshioka et al. 2003 Biol. Reprod. 69, 2092–2099). Presumptive zygotes were cultured in porcine zygote medium (PZM)-5 (Suzuki et al., 2002) containing 2 mM of glutamine and 5 mM of hypotaurine as a basal culture medium until Day 2. The cleaved embryos were then transferred into one of four media prepared as follows: (1) containing no glutamine or hypotaurine (G−H−), (2) containing glutamine (G+H−), (3) containing hypotaurine (G−H+), (4) containing glutamine and hypotaurine (G+H+) (= PZM-5), and cultured for 24 h. After culture, the total number of cells, intracellular GSH content, and level of hydrogen peroxide (H2O2), which is a reactive oxygen species, in the cleaved embryos were examined. Some cleaved embryos were cultured in PZM-5 from Day 3 until Day 5 and the percentage of embryos that developed into blastocysts and the total number of cells in the blastocysts were investigated. Intracellular GSH content and H2O2 level on Day 3 were determined by a dithionitrobenzoic acid-glutathione disulfide (DTNB-GSSG) reductase recycling assay and dichlorohydrofluorescein diacetate (DCHFDA)-based assay, respectively. Data were statistically analyzed by ANOVA and Fisher's PLSD test. The total number of cells (4.3 to 4.4 cells) and intracellular GSH content (2.3 to 2.9 pmol/embryo) in the cleaved embryos on Day 3 did not differ among treatments. On Day 3, the intracellular H2O2 level of the cleaved embryos cultured in G+H+ decreased by 49% compared with those cultured in G−H− (100%) (P < 0.05). On Day 5, the percentage of embryos that developed into blastocysts in G+H+ (52%, 47/90) and G+H− (41%, 36/88) was significantly higher than in G−H− (11%, 11/90) and G−H+ (21%, 19/89) (P < 0.05). The total number of cells in the Day 5 blastocysts from G+H+ (34.5 cells) was significantly (P < 0.05) greater than in those from G−H− (25.8 cells). These results suggest that the presence of glutamine and hypotaurine in PZM-5 from Day 2 to Day 3 improves the subsequent development of porcine embryos into blastocysts by reducing intracellular H2O2 levels. This work was supported by MAFF, Japan.

scholarly journals Gelatin Electrospun Mat as a Potential Co-culture System for In Vitro Production of Sperm Cells from Embryonic Stem Cells

2020 ◽  
Vol 6 (10) ◽  
pp. 5823-5832
Author(s):  
Mina Vardiani ◽  
Marefat Ghaffari Novin ◽  
Morteza Koruji ◽  
Hamid Nazarian ◽  
Ellen Goossens ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Culture System ◽  
Embryonic Stem ◽  
Sperm Cells ◽  
In Vitro Production ◽  
Vitro Production
Sign in / Sign up

Export Citation Format

Share Document