Oesophagostomum radiatum: inhibition of in vitro development by newborn calf serum

1990 ◽  
Vol 64 (1) ◽  
pp. 9-14
Author(s):  
I. J. East ◽  
C. J. Fitzgerald
Keyword(s):  
Inhibitory Action ◽  
Natural Infection ◽  
Calf Serum ◽  
Foetal Calf Serum ◽  
Newborn Calf Serum ◽  
In Vitro Development ◽  
Specific Antibodies ◽  
Serum Inhibition ◽  
Vitro Development

ABSTRACTOesophagostomum radiatum developed to fourth stage larvae after 14 days in in vitro culture. However, development was totally inhibited if the standard 50% foetal calf serum in the medium was replaced by newborn calf serum. Inhibition did not occur with serum from cattle immune to O. radiatum through natural infection or experimental vaccination irrespective of the titre of specific antibodies to O. radiatum in each serum. The inhibitory action of NCS could be abolished by heat treatment at 56°C for 1 h but not by dialysis or repeated freeze-thawing. The inhibition was not consistent with observed differences in the activity of 19 enzymes in the various sera or the absence of various thiol-containing stimulants of worm development.

scholarly journals 138 THE EFFECT OF SYNTHETIC HYALURONAN, BSA AND SERUM ON IN VITRO DEVELOPMENT AND GENE TRANSCRIPTION OF BOVINE BLASTOCYSTS

2005 ◽  
Vol 17 (2) ◽  
pp. 219
Author(s):  
A. Gutiérrez-Adán ◽  
H. Rodriguez-Martinez ◽  
P. Beltrán Breña ◽  
J. De la Fuente ◽  
A.T. Palasz
Keyword(s):  
Fine Structure ◽  
Repeated Measures ◽  
Expression Patterns ◽  
Calf Serum ◽  
The Other ◽  
Bovine Embryo ◽  
In Vitro Development ◽  
Chi Square Analysis ◽  
Vitro Development

The objective of this study was to examine the effect of synthetic hyaluronan (s-HA), BSA and fetal calf serum (FCS) on bovine embryo in vitro development, ultrastructure, and mRNA transcription of four developmentally important genes: apoptosis (BAX), oxidative stress (SOX), growth factor (IGF-II), and cell-to-cell adhesion (Ecad). A total of 1406 presumptive zygotes (7 replicates) were cultured initially in two Groups: 1, SOFaa + 4% BSA only, and 2, SOFaa + 4% BSA and 10% FCS. On Day 4 (96 h after insemination) of culture, the number of zygotes that developed to the <8-cell-stage were recorded, and 2.5 mg/mL of s-HA (MAP-5; Bioniche Inc, Belleville, ON, Canada) was added to half of the embryos from each group; the other half received an equivalent volume of corresponding SOF medium without s-HA. Embryos were cultured in 50-μL drops (25 zygotes per drop) under paraffin oil at 39°C and 5% CO2 in humidified air. Cleavage rates were recorded on Day 2 and the number of blastocysts on Days 7, 8, and 9. At least five blastocysts from each replicate and from each treatment were frozen for evaluation of gene expression patterns. Poly(A) mRNA was prepared from 4–5 groups of pools of 10 embryos. The quantification of all gene transcripts was performed by real time quantitative RT-PCR in three replicates. The fine structure of blastocysts was studied using transmission electron microscopy. Embryo developmental stages and blastocyst formation were analyzed by chi-square analysis, and data on mRNA expression were analyzed by one-way repeated-measures ANOVA. No differences in cleavage rates were observed between groups. There was no difference between the BSA group with or without s-HA in the percentages of embryos developed to the blastocyst stage at Days 7, 8, and 9 (38.3 and 38.1%, respectively). However, significantly (P < 0.05) less blastocysts developed in medium supplemented with BSA + FCS (18.3%) or with BSA + FCS + s-HA (27.4%). Synthetic HA added to the medium containing BSA significantly (P < 0.05) increased the level of expression of EGF-II and decreased (P < 0.05) the level of expression of BAX, SOX, and Ecad. On the other hand, presence of FCS significantly (P < 0.05) increased the level of SOX and decreased the level of IGF-II (P < 0.05), and the addition of s-HA to SOF containing FCS showed no effect on the level of transcription of any analyzed genes. The general fine structure of embryos cultured with s-HA regardless of protein supplement was conspicuously improved in comparison with the respective controls. It can be concluded that, within our culture system, addition of s-HA on Day 4 of culture to the SOFaa medium supplemented with BSA but not in combination with FCS showed a positive effect on embryo development and molecular composition of the embryos.

scholarly journals 25COLD STORAGE OF TISSUES AS SOURCE FOR DONOR CELLS DOES NOT REDUCE THE IN VITRO DEVELOPMENT OF BOVINE EMBRYOS FOLLOWING NUCLEAR TRANSFER

2004 ◽  
Vol 16 (2) ◽  
pp. 135 ◽  
Author(s):  
S. Arat ◽  
H. Bagis ◽  
F. Ergin ◽  
H. Sagirkaya ◽  
H.O. Mercan ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cell Line ◽  
Nuclear Transfer ◽  
Calf Serum ◽  
Calcium Ionophore ◽  
Penicillin G ◽  
In Vitro Development ◽  
Viable Cells ◽  
Vitro Development

So far, most calves have been cloned from live adult cows or fresh fetal samples. There are few reports on using cells from a dead mammal for nuclear transfer (NT). This study was conducted to investigate whether different kind of viable cells could be obtained from tissues stored in cold for different duration and whether these cells could be used for NT. Bovine oocytes isolated from slaughterhouse ovaries were matured in TCM199 supplemented with 10% fetal calf serum (FCS), 50μgmL−1 sodium pyruvate, 1% v:v penicillin-streptomycin (10.000UmL−1 penicillin G, 10.000μgmL−1 streptomycin), 10ngmL−1 EGF, 0.5μgmL−1 FSH, and 5μgmL−1LH. First cell line (CC) was established from articular cartilage of the leg of a slaughtered cow stored at 0°C in a cold storage room for 48h. Second cell line (MC) was established from leg muscle of a cow carcass stored at 0°C for 24h. Tissues from articular cartilage and muscle were cut into small pieces. Tissue explants were cultured in DMEM-F12 supplemented with 10% FBS at 37°C in 5% CO2 in air. Bovine granulosa cells (GC) were isolated from ovarian follicles and used for NT as control cells. Prior to NT, all somatic cells were allowed to grow to confluency (G1/G0) in DMEM-F12 supplemented with 10% FBS. Cumulus cells were removed by vortexing with hyaluronidase at 18h after the start of maturation. Matured oocytes labeled with DNA fluorochrome Hoechst 33342 were enucleated under UV to ensure full removal of the chromatin. A single cell was inserted into the perivitelline space of the enucleated oocyte. Oocyte-cell couples were fused by a DC pulse of 133V/500μm for 25μs. After fusion, NT units were activated using a combination of calcium ionophore (5μM), cytochalasin D (2.5μgmL−1), and cycloheximide (10μgmL−1), and cultured for 7 days. Differences among groups were analyzed by one-way ANOVA after arcsin square transformation. The results are summarized in Table 1. The results suggest that viable cells can be obtained from articular cartilage and muscle of a cow carcass stored at cold temperature for 24 and 48h and these cells have ability to generate NT blastocysts at rates similar to that of the controls. This study was supported by a grant from TUBITAK, Turkey (VHAG-1908-102V048). F Ergin is a volunteer young researcher. Table 1 In vitro development of NT embroys from different cell lines

scholarly journals 143 IN VITRO DEVELOPMENT OF OVINE OOCYTES FROM EWES OF CONTRASTING VITAMIN B12 STATUS

2005 ◽  
Vol 17 (2) ◽  
pp. 222
Author(s):  
L.M. Mitchell ◽  
G. McCallum ◽  
K. Mackie ◽  
M. Ewen ◽  
A. Ainslie ◽  
...  
Keyword(s):  
Vitamin B12 ◽  
Calf Serum ◽  
Cumulus Cells ◽  
Developmental Competence ◽  
Culture Period ◽  
Deficient Diet ◽  
In Vitro Development ◽  
Cell Numbers ◽  
Vitro Development

Suboptimal circulating concentrations of vitamin B12 are commonly found in cattle and sheep grazing cobalt-deficient pastures. Vitamin B12 is a co-factor for enzymes involved in energy metabolism (methylmalonyl CoA mutase) and DNA synthesis/methylation (methionine synthase), and vitamin B12 status may therefore impact on cell division and gene expression in early embryos. The aim of this study was to determine the effect of vitamin B12 status on the in vitro development of ovine oocytes to the blastocyst stage. Mature Scottish Blackface ewes from cobalt-deficient farms were housed for ∼ four months and fed a cobalt-deficient diet (0.06 mg cobalt kg DM−1). At housing, 55 of the ewes were given an intra-ruminal slow-release cobalt bolus to compensate for the dietary deficit, and 52 remained untreated. The ovaries of all ewes were recovered at slaughter within the natural breeding season. Oocytes were aspirated and those with evenly granulated cytoplasm and >3 layers of cumulus cells were pooled according to ewe cobalt treatment, matured, fertilized, and cultured in vitro (∼20 oocytes per 50-μL drop under mineral oil). Oocytes were matured for 24 h in M199 + 10% fetal calf serum at 38.5°C in a humidified atmosphere of 5% CO2 in air prior to co-incubation for 18 h with frozen-thawed semen from a single ejaculate (1 × 106 live sperm mL−1). Presumptive zygotes were cultured for 7 Days in synthetic oviduct fluid + 0.4% fatty acid-free BSA (5% CO2, 5% O2, 90% N). Blastocysts formed at the end of the culture period were fixed and stained (Hoechst 33258) to count cell numbers. Data were analyzed by ANOVA and chi-square. For cobalt-supplemented and non-supplemented ewes, circulating concentrations of vitamin B12 at the time of slaughter were 1244 ± 52.5 and 372 ± 27.9 pmol L−1 (P < 0.001), respectively. Numbers of small (<5 mm) follicles per ewe were 17.6 ± 1.22 and 17.1 ± 1.31, and large (>5 mm) follicles per ewe were 1.8 ± 0.16 and 1.6 ± 0.18, respectively (NS). Cobalt-supplemented ewes yielded a lower proportion of matured oocytes that cleaved but an increased proportion of cleaved oocytes that formed blastocysts by Day 6 of the culture period (Table 1). The proportion of grade 1 and 2 blastocysts was also increased but cobalt treatment did not affect blastocyst cell numbers. In conclusion, results suggest that cleaved eggs derived from ewes of adequate, compared to suboptimal vitamin B12 status have improved developmental competence in vitro. Table 1. Effect of cobalt/vitamin B12 status on the in vitro development of ovine oocytes This work was funded by the Scottish Executive Environment and Rural Affairs Department.

In vitro development of human triploid zygotes reconstructed by pronuclear transfer

2002 ◽  
Vol 78 ◽  
pp. S180-S181
Author(s):  
John Zhang ◽  
Yi Ming Shu ◽  
Lewis C Krey ◽  
Hui Liu ◽  
Guang Lun Zhuang ◽  
...  
Keyword(s):  
In Vitro Development ◽  
Pronuclear Transfer ◽  
Vitro Development

Gallic acid promotes the in vitro development of sheep secondary isolated follicles involving the phosphatidylinositol 3-kinase pathway

2021 ◽  
pp. 106767
Author(s):  
Gizele A.L. Silva ◽  
Luana B. Araújo ◽  
Larissa C.R. Silva ◽  
Bruna B. Gouveia ◽  
Ricássio S. Barberino ◽  
...  
Keyword(s):  
Gallic Acid ◽  
Phosphatidylinositol 3 Kinase ◽  
In Vitro Development ◽  
Vitro Development ◽  
Kinase Pathway ◽  
Phosphatidylinositol 3

scholarly journals Necrostatin-1 Supplementation to Islet Tissue Culture Enhances the In-Vitro Development and Graft Function of Young Porcine Islets

2021 ◽  
Vol 22 (16) ◽  
pp. 8367
Author(s):  
Hien Lau ◽  
Shiri Li ◽  
Nicole Corrales ◽  
Samuel Rodriguez ◽  
Mohammadreza Mohammadi ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Oxygen Consumption ◽  
Insulin Secretion ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
In Vitro Development ◽  
Porcine Islets ◽  
Vitro Development

Pre-weaned porcine islets (PPIs) represent an unlimited source for islet transplantation but are functionally immature. We previously showed that necrostatin-1 (Nec-1) immediately after islet isolation enhanced the in vitro development of PPIs. Here, we examined the impact of Nec-1 on the in vivo function of PPIs after transplantation in diabetic mice. PPIs were isolated from pancreata of 8–15-day-old, pre-weaned pigs and cultured in media alone, or supplemented with Nec-1 (100 µM) on day 0 or on day 3 of culture (n = 5 for each group). On day 7, islet recovery, viability, oxygen consumption rate, insulin content, cellular composition, insulin secretion capacity, and transplant outcomes were evaluated. While islet viability and oxygen consumption rate remained high throughout 7-day tissue culture, Nec-1 supplementation on day 3 significantly improved islet recovery, insulin content, endocrine composition, GLUT2 expression, differentiation potential, proliferation capacity of endocrine cells, and insulin secretion. Adding Nec-1 on day 3 of tissue culture enhanced the islet recovery, proportion of delta cells, beta-cell differentiation and proliferation, and stimulation index. In vivo, this leads to shorter times to normoglycemia, better glycemic control, and higher circulating insulin. Our findings identify the novel time-dependent effects of Nec-1 supplementation on porcine islet quantity and quality prior to transplantation.

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