scholarly journals Effects of some precursors and elicitors on the growth and α-glucosidase inhibitory activity of in vitro Urena lobata L. hairy roots

2020 ◽  
Vol 4 (3) ◽  
pp. First
Author(s):  
Phuong Thi Bach Vu ◽  
Dai Minh Cao ◽  
Phuong Ngo Diem Quach
Keyword(s):  
Salicylic Acid ◽  
Methyl Jasmonate ◽  
Hairy Roots ◽  
Inhibitory Activity ◽  
Culture Medium ◽  
Culture Media ◽  
Culture Conditions ◽  
Metabolic Factors

Recent studies have demonstrated the potential of in vitro Urena lobata L. hairy roots to inhibit α-glucosidase for supporting the treatment of type 2 diabetes. To increase the productivities of hairy roots with α-glucosidase inhibitory activity in in vitro culture conditions, this study focus on the effects of some metabolic factors such as precursors (L-phenylalanine, L-tyrosine) and elicitors (chitosan, methyl jasmonate, salicylic acid). They were added to the culture media to investigate the growth and α-glucosidase inhibitory activity of Urena lobata L. hairy roots. The results showed that for the effects of precursors, only phenylalanine (1 µM) increased root biomass with the highest of α-glucosidase inhibitory activity on the 25th day of culture. In contrast, tyrosine did not play any role in increasing the biomass and α-glucosidase inhibitory activity in Urena lobata L. hairy roots. For the effects of elicitors, only chitosan (50 mg/L) resulted in hairy roots with α-glucosidase inhibitory activity higher than the control after 3 days in culture medium. Other elicitors such as methyl jasmonate, salicylic acid had lower α-glucosidase inhibitory activity than the control. The results of this study demonstrated the potential of phenylalanine and chitosan in increasing the productivity of in vitro hairy roots with higher α-glucosidase inhibitory activity in Urena lobata L.  

The effect of osmolarity on mouse parthenogenesis

Development ◽  
1974 ◽  
Vol 31 (2) ◽  
pp. 513-526
Author(s):  
M. H. Kaufman ◽  
M. A. H. Surani
Keyword(s):  
Protein Synthesis ◽  
Culture Medium ◽  
Culture Media ◽  
Polar Body ◽  
Amino Acid Mixture ◽  
Culture Conditions ◽  
Follicle Cells ◽  
Acid Mixture ◽  
Second Polar Body

Eggs from (C57B1 × A2G)F1 mice were activated by treatment with hyaluronidase, which removed the follicle cells, and cultured in vitro. Observations were made 6–8 h after hyaluronidase treatment to determine the frequency of activation and the types of parthenogenones induced. Cumulus-free eggs resulting from hyaluronidase treatment were incubated for 2¼ h in culture media of various osmolarities. The frequency of activation was found to be dependent on the postovulatory age of oocytes, while the types of parthenogenones induced were dependent on the osmolarity of the in vitro culture medium and their postovulatory age. Culture in low osmolar medium suppressed the extrusion of the second polar body (2PB). This decreased the incidence of haploid eggs with a single pronucleus and 2PB and immediately cleaved eggs from 97·5% to 42·3% of the activated population. Where 2PB extrusion had been suppressed, 97·4% of parthenogenones contained two haploid pronuclei. Very few were observed with a single and presumably diploid pronucleus. Serial observations from 11 to 18 h after hyaluronidase treatment were made on populations of activated eggs as they entered the first cleavage mitosis after 2¼ h incubation in medium either of normal (0·287 osmol) or low (0·168 osmol) osmolarity. A delay in the time of entry into the first cleavage mitosis similar to the duration of incubation in low osmolar medium was observed. Further, eggs were incubated in control and low osmolar culture media containing uniformly labelled [U-14C]amino acid mixture to examine the extent of protein synthesis in recently activated eggs subjected to these culture conditions. An hypothesis is presented to explain the effect of incubation in low osmolar culture medium in delaying the first cleavage mitosis.

291 EFFECTS OF ANTIOXIDANT AND CULTURE MEDIUM ON THE DEVELOPMENT OF PORCINE IN VITRO-MATURED - IN VITRO-FERTILIZED EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 261
Author(s):  
C. Choe ◽  
D.-S. Son ◽  
S.-H. Choi ◽  
S.-R. Cho ◽  
H.-J. Kim ◽  
...  
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Culture Conditions ◽  
Free Oxygen Radicals ◽  
Blastocyst Formation ◽  
Free Oxygen ◽  
Statistical Analysis System ◽  
Developmental Rates ◽  
Analysis System

Most cells cultured in vitro are exposed to the risk of injury by free oxygen radicals (FOR). However, some of FOR-induced injury could be reduced by the antioxidants and culture medium used for in vitro embryos. This study was undertaken to examine the effects of the antioxidant and culture medium on the development of porcine in vitro-matured–in vitro-fertilized embryos. In Experiment 1, we treated the porcine oocytes in NCSU23 medium with various concentrations of β-mercaptoethanol (β-ME) to determine the effective concentration of antioxidants during IVM of porcine oocytes. In Experiment 2, we tested different culture media to find the proper culture conditions for in vitro porcine embryos. The porcine oocytes that were matured in NCSU23 medium and then fertilized in mTBM medium were cultured in NCSU23 or porcine zygote medium-5. All steps (maturation, fertilization, and development) were carried out in vitro. Differences were analyzed among treatments using the general linear model (GLM) procedure in the Statistical Analysis System (SAS Institute, Inc., Cary, NC, USA). The results were summarized as follows. Various concentrations of β-ME showed different developmental rates in porcine embryos. The rates of blastocyst formation at Day 7 after IVF were 9.2 � 1.8 (n = 65), 10.0 � 4.2 (n = 80), 17.5 � 1.1 (n = 63), 20.7 � 1.7 (n = 82), and 14.6 � 1.4 (n = 82) in oocytes treated with β-ME at 0, 10, 25, 50, and 100 �M during IVM, respectively. Of the concentrations of β-ME tested, 50 �M β-ME markedly increased the rates of blastocyst formation at Day 7 (P < 0.05). The rates of blastocyst formation at Day 7 in the NCSU23 and PZM-5 culture media of porcine IVF-derived embryos were 18.8 � 2.6 (n = 96) and 15.6 � 7.1 (n = 77), respectively. The developmental rates were slightly increased in NCSU23, compared with those in PZM-5, but there were no significant differences (P < 0.05) between the NCSU23 and PZM-5 media. In conclusion, these results suggest that the addition of 50 �M β-ME in the IVM medium can improve developmental the rates of porcine embryos in vitro.

The effect of strain of donor eggs, culture conditions, and experimental protocol on release of eggshell calcium by the chick chorioallantoic membrane in vitro

1997 ◽  
Vol 75 (9) ◽  
pp. 1474-1478 ◽  
Author(s):  
Mary J. Packard
Keyword(s):  
Culture Medium ◽  
Calcium Release ◽  
Culture Media ◽  
Chorioallantoic Membrane ◽  
Culture Conditions ◽  
Factors Affecting ◽  
Chick Chorioallantoic Membrane ◽  
Contrast Exposure ◽  
Donor Eggs

Factors affecting release of calcium by the chick chorioallantoic membrane in vitro were assessed by culturing explants of shell with and without the chorioallantoic epithelium. Explants were removed from eggs on day 16 of incubation and cultured using techniques similar to those used to examine resorption of bone in vitro. The region of an egg from which an explant is removed, the strain of donor eggs used, and addition of bovine serum albumin to the culture medium did not affect release of calcium. In contrast, exposure of explants to a variety of different culture media (DMEM, Medium 199, BGJb, and RPMI) introduced considerable variation in calcium release. Calcium release in vitro was also examined by removing the blunt end of a fertile egg as well as the embryo and yolk. The remaining preparation was filled with culture medium, covered, and cultured for two 24-h periods, with a change of medium after the first 24 h. Preparations from which the chorioallantoic membrane had been removed released very little calcium during culture, whereas those with the epithelium in place released considerable quantities. However, release of calcium by preparations with the membrane declined during the second interval. When surface area is taken into account, such preparations mobilize less calcium than explants cultured under similar conditions.

scholarly journals Improving In Vitro Culture of Human Male Fetal Germ Cells

Cells ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 2033
Author(s):  
Myriam Martin-Inaraja ◽  
Monica Ferreira ◽  
Jasin Taelman ◽  
Cristina Eguizabal ◽  
Susana M. Chuva De Sousa Lopes
Keyword(s):  
Germ Cells ◽  
Culture Medium ◽  
Culture Media ◽  
Culture Conditions ◽  
Pronounced Difference ◽  
Coated Substrate ◽  
Male Gametes ◽  
The Media

Male human fetal germ cells (hFGCs) give rise to spermatogonial stem cells (SSCs), which are the adult precursors of the male gametes. Human SSCs are a promising (autologous) source of cells for male fertility preservation; however, in contrast to mouse SSCs, we are still unable to culture them in the long term. Here, we investigated the effect of two different culture media and four substrates (laminin, gelatin, vitronectin and matrigel) in the culture of dissociated second trimester testes, enriched for hFGCs. After 6 days in culture, we quantified the presence of POU5F1 and DDX4 expressing hFGCs. We observed a pronounced difference in hFGC number in different substrates. The combination of gelatin-coated substrate and medium containing GDNF, LIF, FGF2 and EGF resulted in the highest percentage of hFGCs (10% of the total gonadal cells) after 6 days of culture. However, the vitronectin-coated substrate resulted in a comparable percentage of hFGCs regardless of the media used (3.3% of total cells in Zhou-medium and 4.8% of total cells in Shinohara-medium). We provide evidence that not only the choices of culture medium but also choices of the adequate substrate are crucial for optimizing culture protocols for male hFGCs. Optimizing culture conditions in order to improve the expansion of hFGCs will benefit the development of gametogenesis assays in vitro.

Some features of clonal micropropagation of decorative cultivars of Amelanchier Medik.

2020 ◽  
pp. 97-104
Author(s):  
E. N. Raeva-Bogoslovskaya ◽  
O. I. Molkanova
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Culture Media ◽  
Culture Conditions ◽  
Clonal Micropropagation ◽  
Morphogenetic Potential

In vitro culture conditions were optimized for representatives of the genus Amelanchier Medik. at the stages of micropropagation and rooting. A significant effect of the mineral and hormonal compositions of culture media on the morphogenetic potential of the cultivars of serviceberry has been established. The use at the stage of micropropagation of the MS culture medium with addition 1,0 mg / L 6-benzylaminopurine (6-BAP) promoted the active microshoot regeneration of the studied genotypes. For the induction of rhizogenesis the type of auxin as IB A at a concentration of 1,0 mg / L was used.

scholarly journals The effect of coconut extract on callus growth and ultrasound waves on production of betulin and betulinic acid in in-vitro culture conditions of Betula pendula Roth species

2018 ◽  
Vol 60 (4) ◽  
pp. 261-268
Author(s):  
Vahide Payamnoor ◽  
Razieh Jafari Hajati ◽  
Negar Khodadai
Keyword(s):  
Callus Induction ◽  
Betulinic Acid ◽  
High Performance ◽  
Betula Pendula ◽  
Culture Medium ◽  
Culture Media ◽  
Culture Conditions ◽  
Dichlorophenoxyacetic Acid ◽  
Ultrasound Waves

Abstract To determine the effect of coconut extract on callogenesis of Betula pendula, Roth stem barks were cultured in NT (Nagata and Takebe) basic culture media in two individual experiments: i) cultivation explant in different treatments of coconut extracts combined with 1 mg l−1 2, 4-D (2, 4-Dichlorophenoxyacetic acid) and ii) callogenesis in NT media containing 1.5 mg l−1 2,4-D and 0.5 mg l−1 BAP (6-Benzylaminopurine) and then cultivation under the first experiment treatments. The first experiment demonstrated that not all concentrations of coconut extracts lead to callus induction individually, but callus induction increased 84% in a culture containing 5% coconut extract plus 1 mg l−1 2, 4-D. Based on the results of the second experiment, this treatment also significantly increased the wet and dry weights of the produced calluses. The possibility of increasing the betulinic acid and betulin by ultrasound was also studied. Samples cultivated in the selected culture medium were exposed to ultrasound waves in two forms of 1) one exposure and 2) twice exposure (repetition with 24 hr interval) in steps of 20, 60, 100, and 160 sec, and one treatment as the control. Betulin and the betulinic acid amount were determined using HPLC (High-Performance Liquid Chromatography). The maximum betulinic acid and betulin were obtained in 100 sec in once exposure (2.3 mg g−1 DW) and 160 sec in the twice exposure (0.75 mg g−1 DW) to ultrasound. The results showed that betulinic acid content was more than betulin.

Pertumbuhan Dan Perkembangan Anggrek Dendrobium anosmum Pada Media Kultur In Vitro Dengan Beberapa Konsentrasi Air Kelapa

Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Water Concentration ◽  
Medium Composition ◽  
Coconut Water ◽  
Orchid Species ◽  
Randomized Design ◽  
Wet Weight ◽  
Completely Randomized Design

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.

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 Establishment of Highly Efficient Agrobacterium Rhizogenes-mediated Transformation for Stevia Rebaudiana Bertoni Explants

2016 ◽  
Vol 58 (1) ◽  
pp. 113-118 ◽  
Author(s):  
Żaneta Michalec-Warzecha ◽  
Laura Pistelli ◽  
Francesca D’Angiolillo ◽  
Marta Libik-Konieczny
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Molecular Level ◽  
Stevia Rebaudiana ◽  
Culture Conditions ◽  
Light Conditions ◽  
Inoculum Concentration ◽  
Stevia Rebaudiana Bertoni ◽  
Pcr Method

Abstract Leaves and internodes from Stevia rebaudiana Bertoni plants growing in different conditions were used for transformation with two strains of Agrobacterium rhizogenes: ATCC 15384 and LBA 9402. Hairy roots formation was observed and the percentage of the transformed explants depended on the type of explant, time of inoculation and inoculum concentration. Inoculation of explants from ex vitro and in vitro plants with LBA 9402 strain led to higher efficiency of transformation than inoculation with ATCC 15384 strain. Growth rate of hairy roots in liquid culture was assessed under light and dark conditions. It was found that the growth of hairy roots decreased significantly under light conditions. Transformation of hairy roots growing in different culture conditions was confirmed at the molecular level using PCR method with primers constructed against rolB and rolC genes from A. rhizogenes.

Biochemical markers for pregnancy in the spent culture medium of in vitro produced bovine embryos

2021 ◽  
Author(s):  
Gabriela de Oliveira Fernandes ◽  
Marcella Pecora Milazzotto ◽  
Andrei Antonioni Guedes Fidelis ◽  
Taynan Stonoga Kawamoto ◽  
Ligiane de Oliveira Leme ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Biochemical Markers ◽  
Culture Medium ◽  
Culture Media ◽  
Ionization Mass ◽  
Metabolic Profiles ◽  
Bovine Embryos ◽  
The Media

Abstract The present study aimed to identify biomarkers to assess the quality of in vitro produced (IVP) bovine embryos in the culture media. IVP embryos on Day (D) 5 of development were transferred to individual drops, where they were maintained for the last 48 h of culture. Thereafter, the medium was collected and the embryos were transferred to the recipients. After pregnancy diagnosis, the media were grouped into the pregnant and nonpregnant groups. The metabolic profiles of the media were analyzed via electrospray ionization mass spectrometry, and the concentrations of pyruvate, lactate, and glutamate were assessed using fluorimetry. The spectrometric profile revealed that the media from embryos from the pregnant group presented a higher signal intensity compared to that of the nonpregnant group; the ions 156.13 Da [M + H]+, 444.33 Da [M + H]+, and 305.97 Da [M + H]+ were identified as biomarkers. Spent culture medium from expanded blastocysts (Bx) that established pregnancy had a greater concentration of pyruvate (p = 0.0174) and lesser concentration of lactate (p = 0.042) than spent culture medium from Bx that did not establish pregnancy. Moreover, pyruvate in the culture media of Bx can predict pregnancy with 90.9% sensitivity and 75% specificity. In conclusion, we identified markers in the culture media that helped in assessing the most viable IVP embryos with a greater potential to establish pregnancy.

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