P–115 Supplementation of healthy Sertoli cells into culture media containing follicle stimulating hormone (FSH)/testosterone (T) has no advantage in germ cell maturation

Abstract Study question In nonobstructive azoospermia (NOA) cases, whether supplementation of healthy Sertoli cells (SCs) has an effect on spermatogenic differentiation in culture medium containing FSH/T. Summary answer Expression of Crem and Acrosin increased significantly in both medium with FSH/T and medium with additional healthy SCs but there was no difference between them What is known already In NOA the induction of spermatogonial stem cells (SSCs) proliferation and differentiation has been demonstrated using different culture systems. SCs have vital roles in the regulation of spermatogenesis. Hormonal control of spermatogenesis is through FSH and T activity on SCs. Growth factors secreted by SCs via FSH, stimulate proliferation and colonization of SSCs. Although germ cells do not express androgen receptors, FSH receptors are localized on spermatogonia. It is not clear whether native SCs are sufficient for FSH/T added to the culture medium to be effective in induction of spermatogenesis, and whether supplementation of healthy SCs will increase this activity. Study design, size, duration 34 NOA and 12 obstructive azoospermia (OA) cases were included. Testicular tissue samples were taken with testicular sperm extraction (TESE) in the study and control groups. In a group of fertile cases, healthy Sertoli cells were identified and purified and then cryopreserved. Tissue samples of each case prepared in standard DMEM/F12 medium were processed in 2 separate environments containing FSH/T and FSH/T plus thawed healthy SCs for 7 days. Participants/materials, setting, methods The characterization of healthy SCs isolated from fertile cases was done by flow cytometry (FC) and immunohistochemistry using antibodies specific for GATA4 and vimentin. FITC-conjugated annexin V/PI staining and MTT assay were performed to compare the viability and proliferation of SCs before and after freezing. FC was used to measure the 7th day levels of specific markers expressed in spermatogonia (Vasa), meiotic cells (Crem) and post-meiotic cells (Protamine–2 and Acrosin). Main results and the role of chance In Annexin V staining, no difference was found in percentages of live and apoptotic SCs, and MTT exhibited that cryopreservation didn’t inhibite the SCs proliferation compared to the pre-freezing state. Vasa and Acrosin basal levels were found to be lower in infertile patients compared to the control group (8.2% vs. 30.6% and 12.8% vs. 30.5%, p < 0.05). Compared to day 0 measurements, on the 7th day in FSH/T environment, Crem level increased by 58.8% and Acrosin level increased by 195.5% (p < 0.05). Similarly, in medium supplemented with healthy SCs, by day 7, the Crem and Acrosin levels were increased to 92.2% and 204.8%, respectively (p < 0.05). Although Vasa and Protamine levels increased in both groups, they did not reach a significant level. No significant difference was found between the 7th day increase rates of Crem, Vasa, Acrosin and Protamine–2 in either FSH/T-containing medium or in medium additionally supplemented with healthy SCs (58.8% vs. 92.2%, 120.6% vs. 79.4%, 195.5% vs. 204.8% and 232.3% vs. 198.4%, respectively, p > 0.05). Our results suggest that freezing-thawing process would not impair the viability and proliferation of SCs, and adding healthy SCs to the culture medium to correct impaired gene expression does not have an advantage over FSH/T. Limitations, reasons for caution The 7-day culture period we determined might be not sufficient for spermatogenic differentiation completion. This period could be extended in order to see further morphological differentiation may need. Wider implications of the findings: The failure of the culture media containing FSH/T to show the expected effectiveness could be thought to be due to the SCs’ inadequate response to these hormones. Therefore, healthy SCs supplementation would be needed, but this could pose ethical issues. Our findings show that it is not necessary. Trial registration number 214S532

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160 COMPARATIVE STUDY OF IN VITRO CULTURE MEDIA AND ASSISTED HATCHING TECHNIQUES ON MOUSE EMBRYOS

Reproduction Fertility and Development ◽

10.1071/rdv29n1ab160 ◽

2017 ◽

Vol 29 (1) ◽

pp. 188

Author(s):

N. C. Negota ◽

L. P. Nethenzheni ◽

M. L. Mphaphathi ◽

D. M. Barry ◽

T. L. Nedambale

Keyword(s):

In Vitro Culture ◽

Chorionic Gonadotropin ◽

Culture Medium ◽

Culture Media ◽

Assisted Hatching ◽

Hatching Rate ◽

Significant Difference ◽

Equine Chorionic Gonadotropin ◽

F1 Generation

The in vitro culture media and assisted hatching techniques remain challenging obstacles to be utilised widely. Mechanical, chemical, enzymatic thinning, and laser-assisted techniques have been used previously but information is still lacking on its application in livestock. The aim of this study was to compare the effect of 2 in vitro culture media (Hamster F10 and TMC-199) and 4 (mechanical, chemical, enzymatic, and laser) assisted hatching techniques on blastocyst formation and hatching rate using murine embryos as a model. The C57/b and Balb/c breeds were raised until they reached maturity and bred naturally to produce F1 generation. The light in the breeding house was controlled at 14 h light and 10 h dark. Feed and water were provided ad libitum for the mice. Superovulation of females were stimulated using equine chorionic gonadotropin and human chorionic gonadotropin. The F1 generation was used for the collection of the 400 blastocysts and randomly allocated into 4 assisted hatching techniques. Blastocysts were paired into a group of 10 and replicated 4 times for each assisted hatching technique. The general linear model of SAS version 9.4 (SAS Institute Inc., Cary, NC, USA) was used to analyse the data. Assisted hatching techniques of laser, mechanical, enzymatic, and chemical yielded 46.9 ± 37.1, 51.1 ± 40.2, 39.1 ± 35.8, and 33.3 ± 4.5%, respectively, under in vitro culture of Hamster F10. The TCM-199, laser, mechanical, enzymatic, and chemical assisted hatching techniques yielded 56.3 ± 43.3, 52.6 ± 35.5, 49.2 ± 37.5, and 33.9 ± 35.5%, respectively, with a significant difference. There was no significant difference observed in assisted hatching techniques and Hamster F10 culture medium. However, the hatching rate of embryos for all techniques was higher when in vitro cultured in TCM than cultured in Hamster F10. Hatching rate of blastocysts increased from chemical, enzymatic, mechanical, and laser with response to Hamster F10 and TCM; thus, laser is a suitable assisted hatching technique with TCM-199.

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INFLUENCE OF CULTURE MEDIUM ON THE SPORULATION AND VIABILITY OF ASPERGILLUS SPP. AND TALAROMYCES SPP. ENTOMOPATHOGENIC FUNGI

JURNAL HAMA DAN PENYAKIT TUMBUHAN TROPIKA ◽

10.23960/j.hptt.11812-22 ◽

2018 ◽

Vol 18 (1) ◽

pp. 12

Author(s):

Yuyun Fitriana ◽

Radix Suharjo ◽

I Gede Swibawa ◽

Purnomo . ◽

Puji Lestari ◽

...

Keyword(s):

Entomopathogenic Fungi ◽

Culture Medium ◽

Culture Media ◽

Spore Production ◽

Corn Meal ◽

Spore Viability ◽

Randomized Design ◽

Honestly Significant Difference ◽

Significant Difference ◽

Completely Randomized Design

Influence of Culture Medium on the Sporulation and Viability of Aspergillus spp. and Talaromyces spp. Entomopathogenic Fungi. The purpose of this study was to determine the effect of three kinds of cultures media on the spore production and viability of Aspergillus spp. (AS1, 6, 7, 9) and Talaromyces spp. (AS2–5, 8, 10) entomopathogenic fungi. This study was arranged using Factorial-Completely Randomized Design (CRD) with 2 factors and 3 replications. The first factor was three kinds of cultures media (potato dextrose agar (PDA), corn meal agar (CMA), and sabouraud dextrose agar (SDA)) and the second one was isolates of Aspergillus spp. Or Talaromyces spp.. Data of spore production and spore viability were tested using ANOVA and if there was significantly difference, the data then further analyzed using Tukey‘s Honestly Significant Difference (HSD) test at 5% of significant level. The spore production of Aspergillus spp. were in the range of 0.58 - 14.27 x 108 spores mL-1 (PDA); 0.28 – 2.68 x 108 spores mL-1 (SDA) and 1.85 - 5.33 x 108 spores mL-1 (CMA). The highest spore production was achieved by AS1 isolate that was grown on PDA media. The spore produced by Talaromyces spp. were in the range of 2.15 – 28.62 x108 spores mL-1 (PDA); 0.28 – 29.43 x108 spores mL-1 (SDA); and 1.88 – 16.63 x108 spores mL-1 (CMA). The highest spore production was produced by AS8 isolate which were cultured on PDA. The spore viability among isolates of the two entomopathogenic fungi were not significantly different. The spore viability of Aspergillus spp. was in the range of 95.10 – 97.66% (PDA), 94.02 – 98.45% (SDA) and 92.86 – 98.20% (CMA). The spore viability of Talaromyces spp. was in the range of 95.83 – 100% (PDA), 85.83 – 100% (SDA), and 90.75 – 100% (CMA). Culture medium influenced spore production but not the spore viability. The best culture media used for spore production of both of the entomopathogenic fungi was PDA media.

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Aquaculture biological waste as culture medium to cultivation of Ankistrodesmus gracilis (Reinsch) Korshikov

Brazilian Journal of Biology ◽

10.1590/1519-6984.175432 ◽

2017 ◽

Vol 78 (3) ◽

pp. 579-587 ◽

Cited By ~ 2

Author(s):

L. H. Sipaúba-Tavares ◽

T. Florêncio ◽

B. Scardoeli-Truzzi

Keyword(s):

Chlorophyll A ◽

Cell Density ◽

Total Phosphorus ◽

Biochemical Composition ◽

Eichhornia Crassipes ◽

Culture Medium ◽

Culture Media ◽

Conventional Culture ◽

Significant Difference ◽

Biological Wastes

Abstract Current study investigated the effectiveness of different macrophytes as culture media for Ankistrodesmus gracilis in laboratory conditions. Significant difference (p < 0.05) was reported in cell density with regard to conventional culture medium (CHU12) and macrophytes culture media. Mean cell density in NPK, Eichhornia crassipes and E. azurea media was higher (p < 0.05) than in conventional culture medium. Chlorophyll-a was higher than 1 g.L-1, except in CHU12 (0.7 ± 0.4 g.L-1) and T. domingensis (0.8 ± 0.3 g.L-1) media. Nitrate decreased sharply as from the 7th-day of the experiment. Ammonium and total phosphorus were highest in culture media and ranged between 0.4 g.L-1 (P. cordata medium) and 1.7 g.L-1 (CHU12 medium) for ammonium, and between 0.8 g.L-1 (CHU12 medium) and 1.9 g.L-1 (T. domingensis medium) for total phosphorus. Results revealed inorganic fertilizer and macrophytes combined with vitamins may be effective as culture media and strongly supports the growth of Ankistrodesmus gracilis, since cell density and biochemical composition are similar to or higher than conventional culture medium (CHU12). Macrophyte is a tool for aquaculture since biological wastes may be used with nutrients to improve the cultivation of microalgae.

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Evaluación de un medio de cultivo para el aislamiento selectivo de campylobacters termotolerantes

Revista ECIPeru ◽

10.33017/reveciperu2014.0004/ ◽

2018 ◽

pp. 26-30

Keyword(s):

Campylobacter Jejuni ◽

Culture Medium ◽

Culture Media ◽

Campylobacter Coli ◽

Selective Isolation ◽

Basic Fuchsin ◽

Average Growth ◽

San Lorenzo ◽

Significant Difference ◽

Microbial Agents

Evaluación de un medio de cultivo para el aislamiento selectivo de campylobacters termotolerantes Evaluation of a culture medium for the selective isolation of thermotolerant campylobacter Álvaro Tresierra-Ayala, Juan Huanaquiri, Ramsés Perea, María Bendayán. Laboratorio de Microbiología. Centro de Investigación de Recursos Naturales, Universidad Nacional de la Amazonía Peruana. Psje. Los Paujiles S/N, San Lorenzo, distrito de San Juan Bautista, Iquitos-Perú. DOI: https://doi.org/10.33017/RevECIPeru2014.0004/ Resumen Se evaluó la capacidad de aislamiento selectivo de un medio de cultivo para campylobacters termotolerantes, el cual fue suplementado con sangre de porcino y agentes restrictivos (sales biliares, tioglicolato de sodio, fucsina básica), en lugar de la sangre de ovino o equino y diversos antibióticos que suelen ser componentes de los medios comúnmente empleados. Se logró aislar e identificar 20 cepas (13 de Campylobacter jejuni y 7 de Campylobacter coli) de un total de 70 muestras, determinándose que no había diferencia estadísticamente significativa respecto a la capacidad de aislamiento entre el medio de cultivo evaluado y el medio Skirrow modificado. Para el análisis comparativo de la cuantificación del crecimiento de las cepas en ambos medios, se empleó la técnica de Miles y Misra modificada [1] y se observó que las cepas no mostraban diferencia estadísticamente significativa en cuanto a su crecimiento en los medios ensayados, obteniéndose recuentos promedios de 23,4 x106 y 21,7x106 UFC/ml, para la especie de C. coli en el medio de cultivo evaluado y el medio Skirrow modificado y de 44,6 x106 y 40,1 x106 UFC/ml para la especie C. jejuni en ambos medios de cultivo, respectivamente. El medio de cultivo evaluado permite la misma capacidad de aislamiento selectivo para campylobacters termotolerantes que el medio Skirrow modificado; además, estos agentes microbianos crecen cuantitativamente de modo similar en ambos medios de cultivo. Descriptores: Campylobacter, medio de cultivo, sangre de porcino, sales y colorantes. Abstract The ability of selective isolation of a culture medium for thermotolerant campylobacters, supplemented with pig blood and restrictive agents (bile salts, sodium thioglycollate, basic fuchsin), instead of the blood of sheep or horses and various antibiotics which are often components of the means commonly employed was evaluated. Twenty strains (13 C. coli y 7 C. jejuni) were isolated and identified from a total of 70 samples There was not significant difference with respect to the isolation ability between the culture medium evaluated and the modified Skirrow medium. For the comparative analysis about quantification of the growth of strains in both media, Miles and Misra technique, modified [1], was employed. So, it was determined that the strains showed no significant difference with respect to their growth on both media studied. Average growth of 23,4 x 106 and 21,7 x 106 CFU/ml for C. coli in culture medium evaluated and the modified Skirrow medium, and 44,6 x 106 and 40,1 x 106 CFU/ml for C. jejuni in both culture media, respectively, were detected. Culture medium evaluated allows the same ability selective isolation of thermotolerant campylobacter than modified Skirrow medium; moreover, these microbial agents grow similarly quantitatively in both culture media. Keywords: Campylobacter, culture medium, blood pig, salts and dyes.

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MiR-191-5p Is Upregulated in Culture Media of Implanted Human Embryo on Day Three of Development

10.21203/rs.3.rs-122465/v1 ◽

2020 ◽

Author(s):

Ricardo Josue Acuña-González ◽

Fela Vanesa Morales-Hernández ◽

Jorge Skiold López-Canales ◽

Jair Lozano-Cuenca ◽

Mauricio Osorio-Caballero ◽

...

Keyword(s):

Embryo Development ◽

Human Embryo ◽

Culture Medium ◽

Culture Media ◽

Expression Profiles ◽

Uterine Cavity ◽

Common Criteria ◽

Significant Difference ◽

Human Embryo Development

Abstract Background: Morphologic features are the most common criteria for selecting human embryo to be transferred to the receptive uterine cavity. However, such characteristics are not valid for embryos in cellular arrest. The aim of this study was to quantify the expression profile of hsa-miR-21-3p, -24-1-5p, -191-5p, and -372-5p on day 3 of culture media from in vitro fertilization (IVF) embryo that were implanted or failed to be implanted in patients (n=25 pregnant and 25, non-pregnant patients). Methods: Fifty patients were accepted in the Department of Reproductive Biology of a Hospital in México City, based on the Institutional inclusion criteria for in vitro fertilization. On day 3 of development, embryos were transferred to women, and the culture medium was collected from implanted embryos (n=25, pregnant patients) and non-implanted embryos (n=25, non-pregnant). In the culture medium, RNA was isolated using TRIzol reagent. MiRNA expression was detected through RT-PCR with specific primers. Expression bands were quantified using an optic density.Results: The expression profiles were compared between pregnant and non-pregnant patients revealing a significant 5.2-fold greater expression of hsa-miR-191-5p in the former group (p ≤0.001) and a significantly higher expression of hsa-miR-24-1-5p (p =0.043) in the latter. No significant difference was found between the two groups in regard hsa-miR-21-3p or hsa-miR-372-5p (p =0.41). Conclusions: According to the results, has-miR-191-5p could possibly be a possible biomarker of adequate human embryo development. This miRNA modulated IGF2BP-1 and IGF2R, which are associated with the implantation window. On the other hand, hsa-miR-24-1-5p may be related to a poor prognosis of human embryo development.

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Follicle-stimulating hormone-dependent estrogen secretion by rat Sertoli cells in vitro: Modulation by calcium

Acta Endocrinologica ◽

10.1530/acta.0.1250280 ◽

1991 ◽

Vol 125 (3) ◽

pp. 280-285 ◽

Cited By ~ 6

Author(s):

J. Alan Talbot ◽

Ann Lambert ◽

Robert Mitchell ◽

Marek Grabinski ◽

David C. Anderson ◽

...

Keyword(s):

Sertoli Cells ◽

Culture Medium ◽

Follicle Stimulating Hormone ◽

Dibutyryl Camp ◽

Immature Rat ◽

Estradiol Secretion ◽

Old Rats ◽

Stimulating Hormone

Abstract We have investigated the role of Ca2+ in the control of FSH-induced estradiol secretion by Sertoli cells isolated from 8-10 days old rats. Exogenous Ca2+ (4-8 mmol/1) inhibited FSH-stimulated E2 secretion such that, with 8 mmol/l Ca2+ and FSH (8 IU/l) E2 secretion decreased from 2091±322 to 1480±84 pmol/l (p<0.002), whilst chelation of Ca2+ in the culture medium with EGTA (3 mmol/l) increased E2 secretion from 360±45 to 1242±133 pmol/l) in the absence of FSH. Further, EGTA (3 mmol/l) markedly potentiated FSH (8 IU/l), forskolin (1 μmol/l) and dibutyryl cAMP (1 mmol/l)-stimulated E2 secretion. Addition of the Ca2+ ionophores, ionomycin (2-5 μmol/l) and A23187 (2 μmol/l), inhibited FSH (8 IU/l)-stimulated E2 secretion by >80%. The effect of ionomycin was totally reversible, whereas that of A23187 was irreversible. Ionomycin (5 μmol/l) had no effect on EGTA-induced E2 secretion in the absence of FSH, but reduced EGTA-provoked E2 secretion by 59% in the presence of FSH (8 IU/l). Similarly, forskolin- and dibutyryl cAMP-provoked E2 production was inhibited 46-50% by ionomycin (5 μmol/l). We conclude that FSH-induced E2 secretion from immature rat Sertoli cells is modulated by intra- and extracellular Ca2+.

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

Agrologia ◽

10.30598/a.v1i1.293 ◽

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.

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The Effect of Zingiber officinale on the Spleen Tissue and Antibody Titer of Broiler Chickens

Journal of Morphological Sciences ◽

10.4322/jms.088315 ◽

2019 ◽

Vol 36 (01) ◽

pp. 046-050

Author(s):

Alireza Taghdisi ◽

Sajjad Hejazi

Keyword(s):

Broiler Chickens ◽

Zingiber Officinale ◽

Control Group ◽

White Pulp ◽

Broiler Chicks ◽

Pulp Tissue ◽

Tissue Samples ◽

Serum Factors ◽

Spleen Tissue ◽

Significant Difference

Introduction Increasing the immune system's function of fighting infectious diseases is very important in the poultry industry. Ginger, scientifically known as Zingiber officinale, belongs to the Zingiberaceae family. The use of ginger in the diet of poultry increases serum levels of superoxide dismutase enzymes and glutathione peroxidase, which are considered to be important antioxidant enzymes. The main objective of the present study is to evaluate the effect of ginger on the spleen tissue of broiler chickens. Material and Methods The specimens comprised 2 groups of 20 Ross breed broiler chicks, for 42 days and were then, examined and tested. The diet was supplemented with 1 g/kg of ginger powder from the beginning of the rearing period. Blood samples of the chicks were randomly collected to measure the levels of hemagglutination (HI). The removed spleens were fixed with 10% formalin buffer. The specimens were cut in 5-micron diameters and stained with hematoxylin and eosin. Results and Conclusion There was a statistically significant difference in the mean of HI blood titers between the chicks in the growth period and final period groups (p < 0.05). The white-pulp tissue samples were more clearly seen in the treatment group than in the control group, and also, it was observed that the wall of the central artery of the white pulp was thicker in the ginger-treated group as compared with the control group. The nutritional value of ginger may vary. Thus, it is necessary to investigate the effect of this plant final on weight gain; the serum factors associated with the metabolic chart, and the response of the immune system to this plant.

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Untargeted Metabolomics Approach for the Discovery of Environment-Related Pyran-2-ones Chemodiversity in a Marine-Sourced Penicillium restrictum

Marine Drugs ◽

10.3390/md19070378 ◽

2021 ◽

Vol 19 (7) ◽

pp. 378

Author(s):

Van-Tuyen Le ◽

Samuel Bertrand ◽

Thibaut Robiou du Pont ◽

Fabrice Fleury ◽

Nathalie Caroff ◽

...

Keyword(s):

Culture Medium ◽

Blue Mussel ◽

Culture Media ◽

Principal Component ◽

Chemical Diversity ◽

Untargeted Metabolomics ◽

High Chemical ◽

Medium Mass ◽

Metabolomics Study ◽

Penicillium Restrictum

Very little is known about chemical interactions between fungi and their mollusc host within marine environments. Here, we investigated the metabolome of a Penicillium restrictum MMS417 strain isolated from the blue mussel Mytilus edulis collected on the Loire estuary, France. Following the OSMAC approach with the use of 14 culture media, the effect of salinity and of a mussel-derived medium on the metabolic expression were analysed using HPLC-UV/DAD-HRMS/MS. An untargeted metabolomics study was performed using principal component analysis (PCA), orthogonal projection to latent structure discriminant analysis (O-PLSDA) and molecular networking (MN). It highlighted some compounds belonging to sterols, macrolides and pyran-2-ones, which were specifically induced in marine conditions. In particular, a high chemical diversity of pyran-2-ones was found to be related to the presence of mussel extract in the culture medium. Mass spectrometry (MS)- and UV-guided purification resulted in the isolation of five new natural fungal pyran-2-one derivatives—5,6-dihydro-6S-hydroxymethyl-4-methoxy-2H-pyran-2-one (1), (6S, 1’R, 2’S)-LL-P880β (3), 5,6-dihydro-4-methoxy-6S-(1’S, 2’S-dihydroxy pent-3’(E)-enyl)-2H-pyran-2-one (4), 4-methoxy-6-(1’R, 2’S-dihydroxy pent-3’(E)-enyl)-2H-pyran-2-one (6) and 4-methoxy-2H-pyran-2-one (7)—together with the known (6S, 1’S, 2’S)-LL-P880β (2), (1’R, 2’S)-LL-P880γ (5), 5,6-dihydro-4-methoxy-2H-pyran-2-one (8), (6S, 1’S, 2’R)-LL-P880β (9), (6S, 1’S)-pestalotin (10), 1’R-dehydropestalotin (11) and 6-pentyl-4-methoxy-2H-pyran-2-one (12) from the mussel-derived culture medium extract. The structures of 1-12 were determined by 1D- and 2D-MMR experiments as well as high-resolution tandem MS, ECD and DP4 calculations. Some of these compounds were evaluated for their cytotoxic, antibacterial, antileishmanial and in-silico PTP1B inhibitory activities. These results illustrate the utility in using host-derived media for the discovery of new natural products.

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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

Polysaccharides ◽

10.3390/polysaccharides2020032 ◽

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.

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