Influence of cAMP modulator supplementation of in vitro culture medium on Bos taurus indicus embryos

2020 ◽  
Vol 141 ◽  
pp. 134-141 ◽  
Author(s):  
Camila Bortoliero Costa ◽  
Paula Alvares Lunardelli ◽  
Patricia Kubo Fontes ◽  
Mateus José Sudano ◽  
Marcelo Fábio Gouveia Nogueira ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Bos Taurus ◽  
Bos Taurus Indicus

Follicular development, morphological integrity, and oxidative stress in bovine preantral follicles cultured in vitro with ascorbic acid

Zygote ◽  
2021 ◽  
pp. 1-7
Author(s):  
Larissa Zamparone Bergamo ◽  
Denis Vinicius Bonato ◽  
Camila Bizarro-Silva ◽  
Francieli Gesleine Capote Bonato ◽  
Tamires Korchovei Sanches ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Ascorbic Acid ◽  
Culture Medium ◽  
Bos Taurus ◽  
Follicular Development ◽  
Ovarian Follicles ◽  
Bos Taurus Indicus ◽  
Bonferroni Test ◽  
And Oxidative Stress

Summary The aim of this study was to evaluate the follicular development, morphological integrity, and oxidative stress of preantral ovarian follicles from Bos taurus indicus females grown in vitro with ascorbic acid. Ovaries (n = 20) from Bos taurus indicus females were collected, fragmented, and were cultured in vitro for 6 or 12 days in minimum essential medium (MEM), or MEM supplemented with 50 or 100 ng/ml ascorbic acid, with an extracellular matrix of agarose gel, in an incubator at 38.5°C; every 2 days, 100% of the culture medium was replaced. The data were analyzed using the chi-squared test and/or Fisher’s exact test. In the event of a significant effect, the proportions were compared using a 2 × 2 proportion test. The oxidative stress analysis data were submitted to analysis of variance followed by the Bonferroni test. Values were considered significant when P ≤ 0.05. The addition of 100 ng/ml of ascorbic acid to the in vitro culture medium of preantral ovarian follicles from bovine females promoted follicular development, was efficient in maintaining morphological integrity, as well as the stability of reactive oxygen species, after 6 days of in vitro culture.

Culture of preantral ovarian follicles of Bos taurus indicus with alpha-lipoic acid

Zygote ◽  
2021 ◽  
pp. 1-7
Author(s):  
Larissa Zamparone Bergamo ◽  
Denis Vinicius Bonato ◽  
Camila Bizarro-Silva ◽  
Francieli Gesleine Capote Bonato ◽  
Suellen Miguez González ◽  
...  
Keyword(s):  
Lipoic Acid ◽  
Culture Medium ◽  
Bos Taurus ◽  
Thiobarbituric Acid ◽  
Ovarian Follicles ◽  
Control Treatment ◽  
Alpha Lipoic Acid ◽  
Antioxidant Power ◽  
Bos Taurus Indicus

Summary The aim of this study was to evaluate follicular development, morphological integrity, and antioxidant potential of preantral ovarian follicles from Bos taurus indicus females grown in vitro with alpha-lipoic acid. Ovaries (n = 24) of Bos taurus indicus (n = 12) females were collected during slaughter and fragmented. A randomly obtained fragment from each pair of ovaries was fixed in Bouin (non-cultivated control; D0). These fragments were intended for classical histology (morphology and evaluation of follicular growth), and a fragment from each pair of ovaries was frozen at −80°C (non-cultivated control; D0), and assigned for analysis of oxidative stress [thiobarbituric acid reactive substances (TBARS), nitroblue tetrazolium (NBT), and ferric reducing antioxidant power (FRAP)]. The remaining fragments were cultured in vitro for 6 (D6) or 12 (D12) days, containing only minimum essential medium (MEM) or MEM supplemented with alpha-lipoic acid (50, 100, or 250 ng/ml), on an extracellular matrix of agarose gel, in an oven at 38.5ºC. Every 2 days, 100% of the culture medium was replaced. Supplementation with 100 ng/ml was effective for maintaining follicular integrity after 6 days of culture (primordial: 51.28%; development: 36.88%; P < 0.0001). There was no difference (P > 0.05) between treatments compared with the non-cultivated control treatment (D0), using the NBT and TBARS assays. Therefore, supplementation of the in vitro culture medium of bovine preantral ovarian follicles with a concentration of 100 ng/ml of alpha-lipoic acid at 6 days of culture was effective for maintaining follicular integrity and, after 6 days, maintaining stable levels of reactive oxygen species.

scholarly journals Rheological and Microstructural Features of Plant Culture Media Doped with Biopolymers: Influence on the Growth and Physiological Responses of In Vitro-Grown Shoots of Thymus lotocephalus

2021 ◽  
Vol 2 (2) ◽  
pp. 538-553
Author(s):  
Natacha Coelho ◽  
Alexandra Filipe ◽  
Bruno Medronho ◽  
Solange Magalhães ◽  
Carla Vitorino ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Culture Media ◽  
Average Pore Size ◽  
Physiological Responses ◽  
Gum Arabic ◽  
Shoot Elongation ◽  
Hydroxyethyl Cellulose ◽  
Plant Culture

In vitro culture is an important biotechnological tool in plant research and an appropriate culture media is a key for a successful plant development under in vitro conditions. The use of natural compounds to improve culture media has been growing and biopolymers are interesting alternatives to synthetic compounds due to their low toxicity, biodegradability, renewability, and availability. In the present study, different culture media containing one biopolymer (chitosan, gum arabic) or a biopolymer derivative [hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC)], at 100 or 1000 mg L−1, were tested regarding their influence on the growth and physiological responses of Thymus lotocephalus in vitro culture. Cellulose-based biopolymers (HEC and CMC) and gum arabic were used for the first time in plant culture media. The results showed that CMC at 100 mg L−1 significantly improved shoot elongation while chitosan, at the highest concentration, was detrimental to T. lotocephalus. Concerning only the evaluated physiological parameters, all tested biopolymers and biopolymer derivatives are safe to plants as there was no evidence of stress-induced changes on T. lotocephalus. The rheological and microstructural features of the culture media were assessed to understand how the biopolymers and biopolymer derivatives added to the culture medium could influence shoot growth. As expected, all media presented a gel-like behaviour with minor differences in the complex viscosity at the beginning of the culture period. Most media showed increased viscosity overtime. The surface area increased with the addition of biopolymers and biopolymer derivatives to the culture media and the average pore size was considerably lower for CMC at 100 mg L−1. The smaller pores of this medium might be related to a more efficient nutrients and water uptake by T. lotocephalus shoots, leading to a significant improvement in shoot elongation. In short, this study demonstrated that the different types of biopolymers and biopolymer derivatives added to culture medium can modify their microstructure and at the right concentrations, are harmless to T. lotocephalus shoots growing in vitro, and that CMC improves shoot length.

scholarly journals Fetal calf serum enhances in vitro production of Bos taurus indicus embryos

2011 ◽  
Vol 75 (3) ◽  
pp. 429-433 ◽  
Author(s):  
F.G. Leivas ◽  
D.S. Brum ◽  
S.S. Fialho ◽  
W.P. Saliba ◽  
M.T.T. Alvim ◽  
...  
Keyword(s):  
Fetal Calf Serum ◽  
Bos Taurus ◽  
Calf Serum ◽  
In Vitro Production ◽  
Bos Taurus Indicus ◽  
Vitro Production

P–218 Analysis of the occurrence of microbial contamination in IVF culture system and the effect of microorganisms on embryo development and clinical outcomes

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
F Du ◽  
R Li ◽  
Q Zhang ◽  
W Wang
Keyword(s):  
Escherichia Coli ◽  
In Vitro Culture ◽  
Clinical Outcomes ◽  
Embryo Transfer ◽  
Follicular Fluid ◽  
Culture System ◽  
Microbial Contamination ◽  
Culture Medium ◽  
Ivf Et

Abstract Study question what is the source, prevalence, and influence of microbial contamination on in vitro fertilization (IVF) and embryo transfer (ET) cycles? Summary answer Microbial contamination mainly occurs on Day 2, most caused by Escherichia coli carried with semen. ICSI could prevent contamination effectively and get good clinical outcomes. What is known already Microbial contamination occurs in IVF-ET system occasionally, which is hard to stop happening. The IVF culture system and laboratory environment, the patients’ follicular fluid and semen are not absolutely sterile, while the antibiotics in culture medium isn’t effective for all microbe types, and the artificial operations may bring in microbes. Generally, microbial contamination leads to degradation of embryos, reduction the number of embryos available, and infection of female reproductive tract, which would increase the cost of patients’ time, money, and bring psychological damages. A better understanding of embryo contamination in IVF culture system is of added value. Study design, size, duration A total of 29583 IVF-ET cycles were enrolled in this prospective observational study, from January 2010 to December 2020, included 70 microbial contamination cycles discovered in Day1-Day3 (D1-D3) of in vitro culture. Follicular fluid and semen saved on oocyte retrieval day, and culture medium contaminated were examined and identified for microorganisms at each contamination cycle. Participants/materials, setting, methods Compared the contamination rate of different insemination methods (IVF/ICSI/IVF+ICSI), different in vitro culture days (D1-D3), and different samples examination (follicular fluid, semen, culture medium) respectively, identified the source of microorganism types, compared the IVF culture outcomes and clinical outcomes between total contamination group (TC group, 42 cases) and partial contamination group (PC group, 28 cases). Main results and the role of chance A total of 70 microbial contamination cases occurred in 29583 oocyte retrieving cycles (0.24%), and it was observed only in IVF embryos but never in ICSI (Intracytoplasmic sperm injection) embryos. 38 contamination cases occurred on D2 with a highest ratio (54.3%) compared to D1 (32.9%) and D3(12.9%); Compared with follicular fluid, semen was the main cause inducing contamination from D1 to D3, and Escherichia coli in semen and culture medium, Enterococcus faecalis in follicular fluid proved to be the most common sources. Compared with TC group, the PC group showed a lower rate of No-available embryos (21.4% vs 81.0%) and a higher rate of blastocyst formation (41.2% vs 28.6%), In addition, the clinical pregnancy rate of PC group was higher than that of TC group in both fresh and frozen-thawed embryo transfer cycles (31.3% vs 16.7%, 38.5% vs 0.0%). Limitations, reasons for caution Further study is still necessary to better understand the sources that induce microbial contamination embryos, and more efficient methods are required to remove the microbes on these contaminated embryos so as better develop and manage a sterile micro-environment for successful embryo growth. Wider implications of the findings: The differential embryonic microbe types associated to different IVF culture and clinical outcomes in patients undergoing IVF-ET might have profound implications for understanding the microbial sources and making a better management of IVF culture system. Trial registration number Not applicable

Synthesis of RNA in oocytes of Xenopus laevis during culture in vitro

Development ◽  
1974 ◽  
Vol 32 (2) ◽  
pp. 515-532
Author(s):  
A. Colman
Keyword(s):  
Xenopus Laevis ◽  
In Vitro Culture ◽  
Ribosomal Rna ◽  
Rna Synthesis ◽  
Culture Medium ◽  
Salt Medium ◽  
Complex Media ◽  
18S Ribosomal Rna ◽  
Rna Precursor

RNA synthesis can be maintained in large oocytes of Xenopus laevis during periods of in vitro culture of at least 10 days. A simple salt medium, modified Barth's solution, is found to be as effective a culture medium for these oocytes as several other complex media. The newly synthesized RNA is characterized electrophoretically and shown to consist predominantly of ribosomal RNA precursor, 28S and 18S ribosomal RNA, and 4S RNA. The distribution of this RNA within the oocyte is detected autoradiographically, where it is found to be greatly concentrated over the nucleoli. No qualitative alterations in either of these parameters are found during culture, within the limits of sensitivity of the assay procedures.

scholarly journals PROPORÇÃO DO SEXO NA PRODUÇÃO IN VITRO DE EMBRIÕES BOVINOS

2015 ◽  
Vol 20 (2) ◽  
Author(s):  
Marcus Vinícius Galvão Loiola ◽  
Alexandra Soares Rodrigues ◽  
Marcos Chalhoub ◽  
Priscila Assis Ferraz ◽  
Rodrigo Freitas Bittencourt ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Bos Taurus Indicus

Objetivou-se avaliar a proporção do sexo utilizando espermatozoides provenientes de sêmen convencional ou sexado para fêmea em um programa de produção in vitro de embriões (PIVE) bovinos. Foram utilizados oócitos de 87 doadoras Bos taurus indicus da raça Nelore, obtidos por sistema de aspiração folicular transvaginal guiado por ultrassonografia. Os oócitos selecionados foram maturados in vitro, fertilizados com espermatozoides oriundos de sêmen convencional (grupo 1 – CONV) ou sêmen sexado para fêmea (grupo 2 – SEX) e os embriões produzidos foram então cultivados in vitro por sete dias. Após o cultivo, os mesmos foram envasados e inovulados em receptoras mestiças Bos taurus taurus x Bos taurus indicus. Foram transferidos 1.319 embriões, sendo 1.081 produzidos com sêmen convencional (CONV) e 238 com sêmen sexado de fêmea (SEX). O diagnóstico precoce de gestação e a sexagem fetal foram realizados por ultrassonografia transretal. A taxa de gestação total foi 37,38% (493/1.319), sendo 37,74% (408/1.081) no grupo CONV e 35,71% (85/238) no grupo SEX. A proporção do sexo no grupo CONV foi desviada (P=0,002) a favor dos machos (57,84% de machos e 42,16% de fêmeas) e no grupo SEX ocorreu um desvio na proporção (P=0,001) a favor das fêmeas (68,24% de fêmeas e 31,76% de machos). Concluiu-se que a utilização do sêmen convencional em PIVE bovinos elevou a proporção de produtos machos e apesar da utilização de sêmen sexado de fêmea ser uma alternativa para pré-selecionar o sexo dos produtos, ainda houve uma considerável taxa de machos quando se utilizou este tipo de sêmen.

scholarly journals A new experimental in vitro culture medium for cultivation of Leishmania species.

1997 ◽  
Vol 35 (9) ◽  
pp. 2430-2431 ◽  
Author(s):  
M E Limoncu ◽  
I C Balcioğlu ◽  
K Yereli ◽  
Y Ozbel ◽  
A Ozbilgin
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Leishmania Species

scholarly journals Morphophysiological responses of Billbergia zebrina Lindl. (Bromeliaceae) in function of types and concentrations of carbohydrates during conventional in vitro culture

2020 ◽  
Vol 26 (1) ◽  
pp. 18-34 ◽  
Author(s):  
Elizangela Rodrigues Santos ◽  
João Paulo Rodrigues Martins ◽  
Luiz Carlos de Almeida Rodrigues ◽  
Andreia Barcelos Passos Lima Gontijo ◽  
Antelmo Ralph Falqueto
Keyword(s):  
Photosystem Ii ◽  
In Vitro Culture ◽  
Leaf Anatomy ◽  
Culture Medium ◽  
Stomatal Density ◽  
Photosynthetic Apparatus ◽  
Fluorescence Transient ◽  
Chlorophyll A Fluorescence Transient ◽  
Concentration Interval

Abstract When propagated in vitro, explants receive all the nutrients needed for their growth, including carbohydrates, from the culture medium. However, it is not well understood how the type and concentration of carbohydrates can affect the functioning of the photosynthetic apparatus (particularly photosystem II) of these plants. The aim was to assess the morphophysiological responses of Billbergia zebrina plants in function of sources and concentrations of carbohydrates during in vitro culture. Side shoots of plants previously established in vitro were individualized and transferred to a culture medium containing fructose, glucose or sucrose in four concentrations (0, 15, 30 or 45 g L−1). After growth for 55 days, the chlorophyll a fluorescence transient, leaf anatomy and growth were analyzed. The concentration and type of carbohydrate employed during in vitro culture did not decrease the photosynthetic apparatus performance. However, concentrations above 30 g L−1 led to anatomical modifications, revealing some degree of stress suffered by the plants. When grown in concentrations of 15 and 30 g L−1, irrespective of the carbohydrate used, the plants presented greater stomatal density. The supplementation of the culture medium with monosaccharides caused alterations in the development of the xylem vessels, such as increased number and diameter, allowing adjustment to the microenvironmental conditions. The in vitro conditions influenced the photosynthetic and anatomical responses of plants. The concentration interval from 15 to 30 g L−1 sucrose had a better effect by not causing large changes in the performance of the photosynthetic apparatus and anatomy of plants.

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