Bacteriophages with Potential to Inactivate Aeromonas hydrophila in Cockles: In Vitro and In Vivo Preliminary Studies

João Duarte; Carla Pereira; Pedro Costa; Adelaide Almeida

doi:10.3390/antibiotics10060710

Bacteriophages with Potential to Inactivate Aeromonas hydrophila in Cockles: In Vitro and In Vivo Preliminary Studies

Antibiotics ◽

10.3390/antibiotics10060710 ◽

2021 ◽

Vol 10 (6) ◽

pp. 710

Author(s):

João Duarte ◽

Carla Pereira ◽

Pedro Costa ◽

Adelaide Almeida

Keyword(s):

Aeromonas Hydrophila ◽

Pathogenic Bacteria ◽

Culture Medium ◽

Phage Therapy ◽

Maximum Reduction ◽

Liquid Culture Medium ◽

Resistant Mutants ◽

Bivalve Shellfish

The recurrent emergence of infection outbreaks associated with shellfish consumption is of extreme importance for public health. The present study investigated the potential application of phages AH-1, AH-4, and AH-5 to inactivate Aeromonas hydrophila, a causative agent of infections in humans associated with bivalve shellfish consumption. The inactivation of A. hydrophila was assessed in vitro, using a liquid culture medium, and in vivo, using artificially contaminated cockles with A. hydrophila ATCC 7966. In the in vitro experiments, all phages were effective against A. hydrophila, but phage AH-1 (with a maximum reduction of 7.7 log colonies forming units CFU/mL) was more effective than phages AH-4 and AH-5 (with reductions of 4.9 and 4.5 log CFU/mL, respectively). The cocktails AH-1/AH-4, AH-1/AH-5, AH-4/AH-5, and AH-1/AH-4/AH-5 were slightly more effective than the single phage suspensions. The phages presented a low emergence rate of phage-resistant mutants. When artificially contaminated cockles were treated in static seawater with phage AH-1, around 44% of the added A. hydrophila (1.0 log CFU/g) was inactivated. The results of this study suggest that phage therapy can be an effective alternative to control human pathogenic bacteria during depuration.

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Influence of genotype on protein and oil concentration of soybean seeds

Brazilian Archives of Biology and Technology ◽

10.1590/s1516-89132010000400007 ◽

2010 ◽

Vol 53 (4) ◽

pp. 793-799 ◽

Cited By ~ 3

Author(s):

Esmael Lopes dos Santos ◽

Antonio Eduardo Pípolo ◽

Ricardo Tadeu de Faria ◽

Cássio Egidio Cavenaghi Prete

Keyword(s):

Protein Concentration ◽

Culture Medium ◽

Seed Oil ◽

Soybean Seeds ◽

Glutamine Concentration ◽

Liquid Culture Medium ◽

Immature Seeds ◽

Oil Concentration

Aiming at evaluating genotype influence on the concentration of protein and oil, immature seeds of cultivars CD 202 and CD 206 were removed from the mother-plant, in the stage R5, and were grown in vitro, in a liquid culture medium which contained 20, 40 and 60 mM of glutamine, during eight days. Afterwards, the concentrations of oil and protein were compared to the contents of the seeds cultivated in vivo. With a higher availability of glutamine for the seed, there was an increase of protein content. The genotypes were statistically different as far as the protein concentration was concerned,which confirmed that the genotype had influence on the concentration of protein in the seed. Oil and protein concentrations were inversely related when a variation of glutamine concentration occurred.

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Screening of Probiotic Bacteria from Intestine and Culture Environment of Hoeven’s slender carp Leptobarbus hoeveni Blkr to Control Pathogenic Bacteria

Jurnal Akuakultur Indonesia ◽

10.19027/jai.9.127-135 ◽

2010 ◽

Vol 9 (2) ◽

pp. 127

Author(s):

. Sunarto ◽

. Sukenda ◽

. Widanarni

Keyword(s):

Aeromonas Hydrophila ◽

Pathogenic Bacteria ◽

Probiotic Bacteria ◽

Streptococcus Iniae ◽

Mycobacterium Fortuitum ◽

Fish Pathogen ◽

Culture Environment ◽

Control Disease

The ability of probiotic bacteria to control disease infection has been used in aquaculture. This experiment was conducted to isolate and characterize probiotic bacteria; the competition test its ability probiotic bacteria against pathogenic bacteria; and to improve survival rate of Leptobarbus hoeveni. The bacteria were isolated from Leptobarbus hoeveni and its culture environment, and then tested to know its ability to inhibit bacterial fish pathogen in-vitro. Furthermore, the selected probiotic bacteria were tested in vivo to evaluate their ability to inhibit pathogen of Leptobarbus hoeveni. The result showed that probiotic bacteria inhibit the growth of Streptococcus iniae, Flexibacter columnaris, Mycobacterium fortuitum and Aeromonas hydrophila in vitro. Isolate DD3 was the best of candidate probiotic because of the ability to inhibit pathogen, especially A. hydrophila, the most virulent bacteria in Leptobarbus hoeveni. Key Words : probiotic bacteria, Leptobarbus hoeveni, pathogenic bacteria Abstrak Kemampuan bakteri probiotik untuk mengendalikan penyakit infeksi telah digunakan dalam akuakultur. Tujuan penelitian ini adalah mengisolasi dan mengkarakterisasi bakteri probiotik, menguji kemampuan bakteri probiotik terhadap bakteri patogen, sehingga dapat meningkatkan tingkat kelangsungan hidup ikan jelawat. Bakteri diisolasi dari usus ikan jelawat dan lingkungan budaya, kemudian diuji kemampuannya menghambat bakteri patogen secara in-vitro. Selanjutnya bakteri probiotik yang dipilih diuji secara in vivo untuk mengevaluasi kemampuannya dalam menghambat patogen di dalam tubuh ikan jelawat. Dari hasil penelitian diperoleh bakteri probiotik yang diisolasi dari usus dan lingkungan budaya ikan jelawat menunjukkan penghambatan pertumbuhan terhadap Streptococcus iniae, Flexibacter columnaris, Mycobacterium fortuitum dan Aeromonas hydrophila secara in vitro. Isolat DD3 merupakan kandidat probiotik terbaik, karena mempunyai kemampuan untuk menghambat bakteri patogen, khususnya bakteri A. hydrophila adalah bakteri yang paling viluren bagi ikan jelawat. Kata Kunci: bakteri probiotik, ikan jalawat dan baktri patogen

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Parallel evolution of phage resistance - virulence trade - offs during in vitro and nasal Pseudomonas aeruginosa phage treatment

10.1101/2021.09.06.459069 ◽

2021 ◽

Author(s):

Meaghan Castledine ◽

Daniel Padfield ◽

Pawel Sierocinski ◽

Jesica Soria Pascual ◽

Adam Hughes ◽

...

Keyword(s):

Pathogenic Bacteria ◽

Evolutionary Dynamics ◽

Phage Therapy ◽

De Novo ◽

Resistant Bacteria ◽

Phage Resistance ◽

Lower Growth ◽

Trade Offs

With rising antibiotic resistance, there has been increasing interest in the treatment of pathogenic bacteria with bacteriophages (phage therapy). One limitation of phage therapy is the ease at which bacteria can evolve resistance. The negative effects of resistance may be partially mitigated when resistance results in reduced bacterial growth and virulence, or when phage coevolve to overcome resistance. Resistance evolution and its consequences are highly contingent on the particular combination of bacteria and phage and the ecological context they interact in, making therapeutic outcomes hard to predict. One solution might be to conduct ″in vitro evolutionary simulations″ using the bacteria-phage combinations specific to the therapeutic context. Here, we investigate parallels between in vitro experiments and in vivo dynamics in a human participant. Evolutionary dynamics were similar in vivo and in vitro, with high levels of de novo resistance evolving quickly with limited evidence of phage evolution. Moreover, resistant bacteria – evolved both in vitro and in vivo – had lower virulence when measured in an insect model. In vivo, this was linked to lower growth rates of resistant isolates, whereas in vitro isolates evolved greater biofilm production with phage resistance. Population sequencing suggests resistance was typically the result of selection on de novo mutations rather than sorting of existing variants in the population. These results highlight the speed at which resistance to phages can evolve in vivo, and that in vitro evolution may give useful insights for evolutionary outcomes in vivo.

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Lipid Secretion by Parasitic Cells of Coccidioides Contributes to Disseminated Disease

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.592826 ◽

2021 ◽

Vol 11 ◽

Author(s):

Carlos Alberto Peláez-Jaramillo ◽

Maria Del Pilar Jiménez-Alzate ◽

Pedronel Araque-Marin ◽

Chiung-Yu Hung ◽

Natalia Castro-Lopez ◽

...

Keyword(s):

Culture Medium ◽

Fungicidal Activity ◽

Saturated Fatty Acids ◽

Outer Wall ◽

Mammalian Host ◽

Liquid Culture Medium ◽

United States Mexico ◽

Disseminated Disease

Coccidioides is a soil-borne fungal pathogen and causative agent of a human respiratory disease (coccidioidomycosis) endemic to semi-desert regions of southwestern United States, Mexico, Central and South America. Aerosolized arthroconidia inhaled by the mammalian host first undergo conversion to large parasitic cells (spherules, 80–100 μm diameter) followed by endosporulation, a process by which the contents of spherules give rise to multiple endospores. The latter are released upon rupture of the maternal spherules and establish new foci of lung infection. A novel feature of spherule maturation prior to endosporulation is the secretion of a lipid-rich, membranous cell surface layer shed in vivo during growth of the parasitic cells and secretion into liquid culture medium during in vitro growth. Chemical analysis of the culture derived spherule outer wall (SOW) fraction showed that it is composed largely of phospholipids and is enriched with saturated fatty acids, including myristic, palmitic, elaidic, oleic, and stearic acid. NMR revealed the presence of monosaccharide- and disaccharide-linked acylglycerols and sphingolipids. The major sphingolipid components are sphingosine and ceramide. Primary neutrophils derived from healthy C57BL/6 and DBA/2 mice incubated with SOW lipids revealed a significant reduction in fungicidal activity against viable Coccidioides arthroconidia compared to incubation of neutrophils with arthroconidia alone. Host cell exposure to SOW lipids had no effect on neutrophil viability. Furthermore, C57BL/6 mice that were challenged subcutaneously with Coccidioides arthroconidia in the presence of the isolated SOW fraction developed disseminated disease, while control mice challenged with arthroconidia alone by the same route showed no dissemination of infection. We hypothesize that SOW lipids contribute to suppression of inflammatory response to Coccidioides infection. Studies are underway to characterize the immunosuppressive mechanism(s) of SOW lipids.

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Application of Bacillus probiotic to prevent Aeromonas hydrophilla infection in Clarias sp.

Jurnal Akuakultur Indonesia ◽

10.19027/jai.13.105-114 ◽

2015 ◽

Vol 13 (2) ◽

pp. 105 ◽

Cited By ~ 3

Author(s):

Mohammad Faizal Ulkhaq ◽

, Widanarni ◽

Angela Mariana Kusumastuti

Keyword(s):

Aeromonas Hydrophila ◽

Pathogenic Bacteria ◽

Clarias Gariepinus ◽

Positive Control ◽

Negative Control ◽

Probiotic Bacteria ◽

African Catfish ◽

Control Application

ABSTRACT The aim of this study was to test the effectiveness of a probiotic Bacillus for the prevention of motile aeromonad septicemia (MAS) disease caused by Aeromonas hydrophila in African catfish (Clarias gariepinus). The study consisted of the inhibition testing of A. hydrophila by Bacillus (in vitro) and the application of probiotic in African catfish (in vivo). The in vivo test, consisted of five treatments such as the addition of probiotic Bacillus P4I1 RifR, Bacillus P4I2 RifR, Bacillus P4I1 RifR + Bacillus P4I2 RifR (Kom), positive control (K+; only added with A. hydrophila) and negative control (K-; without probiotic nor A. hydrophila addition). African catfish (13.35±2.80 g) was maintained in 15 aquariums (40 L in volume) with 30 fishes each for 30 days. Probiotic bacteria was applied in water once a day, whereas pathogenic bacteria A. hydrophila RifR (103 cfu/mL) were added once in earlier treatment (except for the negative control). The result showed that the optimal concentration of Bacillus to inhibit A. hydrophila on in vitro test was 104 cfu/mL. In vivo test showed that the addition of probiotic in media of cultivation could reduce the number of A. hydrophila, improve immune response, and also increase the survival of African catfish compared to positive control. Application of probiotic P4I1 RifR showed the highest survival (92.23%) of all treatments. Keywords: Bacillus, Clarias gariepinus, motile aeromonad septicemia, probiotic ABSTRAK Penelitian ini bertujuan untuk menguji efektivitas probiotik Bacillus dalam pencegahan penyakit motile aeromonad septicaemia (MAS) yang disebabkan oleh Aeromonas hydrophila pada ikan lele dumbo (Clarias gariepinus). Penelitian terdiri atas pengujian penghambatan bakteri probiotik Bacillus terhadap A. hydrophila secara in vitro, dilanjutkan dengan aplikasi pada budidaya ikan lele dumbo (in vivo). Pada uji in vivo, penelitian terdiri atas lima perlakuan yaitu budidaya ikan lele dumbo dengan penambahan probiotik Bacillus P4I1 RifR, Bacillus P4I2 RifR, kombinasi probiotik Bacillus P4I1 RifR + Bacillus P4I2 RifR (Kom), kontrol positif (K+; hanya ditambahkan A. hydrophila) dan kontrol negatif (K-; tanpa pemberian probiotik dan A. hydrophila). Ikan lele dumbo (13,35±2,80 g) dipelihara pada akuarium volume 40 L dengan kepadatan 30 ekor/akuarium selama 30 hari. Bakteri probiotik ditambahkan pada media pemeliharaan ikan setiap hari, sedangkan bakteri patogen A. hydrophila RifR (103 cfu/ mL) diberikan sekali pada awal pemeliharaan (kecuali pada kontrol negatif). Hasil penelitian menunjukkan bahwa konsentrasi terbaik pada penghambatan in vitro adalah dengan penambahan Bacillus 104 cfu/mL. Hasil uji in vivo menunjukkan perlakuan penambahan probiotik pada media budidaya efektif dapat menekan jumlah bakteri A. hydrophila, memperbaiki respons imun, dan meningkatkan kelangsungan hidup ikan lele dumbo dibanding kontrol positif. Perlakuan probiotik P4I1 RifR memberikan hasil terbaik dengan tingkat kelangsungan hidup tertinggi yaitu 92,23%. Kata kunci: Bacillus, Clarias gariepinus, motile aeromonad septicemia, probiotik

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Selection of lactic acid bacteria as a probiotic and evaluated its performance on gnotobiotic catfish Clarias sp.

Jurnal Akuakultur Indonesia ◽

10.19027/jai.17.1.68-80 ◽

2018 ◽

Vol 17 (1) ◽

pp. 68

Author(s):

Enita Romasni Turnip ◽

Widanarni, Widanarni ◽

Anja Meryandini

Keyword(s):

Growth Rate ◽

Lactic Acid ◽

Survival Rate ◽

Lactic Acid Bacteria ◽

Aeromonas Hydrophila ◽

Pathogenic Bacteria ◽

Challenge Test ◽

Feed Conversion

ABSTRACT This study aimed to select lactic acid bacteria (LAB) as a potential probiotic that producing anti‒microbial compounds in order to treat motile aeromonads septicemia diseases caused by Aeromonas hydrophila on catfish Clarias sp. and evaluated its performance on gnotobiotic catfish. The in vitro assay was done to select several LAB isolates based on antagonistic activity against pathogenic bacteria. The selected isolate was tested in vivo to observe their ability to improve growth performances of catfish. The study was conducted with five treatments consists of K‒ (normal catfish without addition probiotic, without challenge test), K+ (normal catfish without addition of probiotic, with challenge test), Np (normal catfish with addition of probiotic and challenge test), G (gnoto catfish without addition of probiotic, with challenge test), and Gp (gnoto catfish with addition of probiotic and challenge test). The results showed that the addition of Pediococcus pentosaceus E2211 as selected probiotic could increase survival rate, specific growth rate, and immune response towards infection of A. hydrophila. The best survival rate after challenge test was obtained in Np and Gp treatments (88.46%), followed by G treatment (65.38%), while the K+ was only 53.84%. The conclusion of this study was P. pentosaceus E2211 potentially used as a probiotic candidate for normal and gnotobiotic catfish. The presence of normal microflora with P. pentosaceus E2211 in Np treatment showed the best probiotic performance with daily growth rate 3.28%, feed conversion ratio 1.79, and total intestinal bacteria reached 108 CFU/mL significantly different from other treatments (P<0.05).Keywords: Aeromonas hydrophila, catfish, LAB, probiotic, screening ABSTRAKTujuan penelitian ini adalah menyeleksi bakteri asam laktat (BAL) sebagai probiotik potensial penghasil senyawa antimikrob guna menanggulangi penyakit motile aeromonad septicemia akibat Aeromonas hydrophila pada ikan lele Clarias sp. dan evaluasi kinerjanya pada ikan lele gnotobiotik. Pengujian in vitro dilakukan untuk menyeleksi beberapa isolat BAL sebagai kandidat probiotik berdasarkan aktivitas antagonis terhadap bakteri patogen. Isolat terpilih kemudian diuji in vivo untuk mengetahui kemampuannya dalam meningkatkan performa tumbuh ikan lele. Penelitian ini menggunakan rancangan acak lengkap dengan lima perlakuan, yaitu: K‒ (lele normal tanpa probiotik dan tanpa tanpa diuji tantang), K+ (lele normal tanpa probiotik dan diuji tantang), Np (lele normal diberi probiotik dan diuji tantang), G (lele gnoto tanpa probiotik dan diuji tantang), dan Gp (lele gnoto diberi probiotik dan diuji tantang). Hasil penelitian menunjukkan pemberian probiotik terpilih BAL Pediococcus pentosaceus E2211 mampu meningkatkan sintasan, laju pertumbuhan, dan respons imun ikan lele terhadap infeksi A. hydrophila. Sintasan terbaik pascauji tantang diperoleh pada perlakuan Np dan Gp yaitu sebesar 88,46%, diikuti perlakuan G sebesar 65,38%, sementara pada K+ hanya mencapai 53,84%. Kesimpulan dari penelitian ini ialah isolat BAL terpilih P. pentosaceus E2211 berpotensi sebagai kandidat probiotik untuk ikan lele normal dan lele gnotobiotik Clarias sp. Keberadaan mikroflora normal yang berasosiasi dengan P. pentosaceus E2211 pada perlakuan Np menunjukkan kinerja probiotik terbaik dengan nilai laju pertubuhan harian 3,28%, rasio konversi pakan 1,79 dan total bakteri usus mencapai 108 CFU/mL yang berbeda signifikan dibanding perlakuan lainnya (P<0,05).Kata kunci: A. hydrophila, BAL, ikan lele, probiotik, seleksi

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PENAMBAHAN KANDIDAT PROBIOTIK Bacillus methylothrophicus SECARA BERKALA PADA MEDIA PEMELIHARAAN UNTUK PENCEGAHAN INFEKSI BAKTERI Aeromonas hydrophila PADA IKAN NILA (Oreochromis niloticus)

Sains Akuakultur Tropis ◽

10.14710/sat.v3i2.4069 ◽

2019 ◽

Vol 3 (2) ◽

Cited By ~ 1

Author(s):

Femy Musthofa Ardy ◽

Desrina Desrina ◽

Alfabetian Harjuno Condro Haditomo

Keyword(s):

Aeromonas Hydrophila ◽

Oreochromis Niloticus ◽

Pathogenic Bacteria ◽

Survival Rates ◽

Average Weight ◽

In Vitro Test ◽

In Vivo Test ◽

Short Period

Aeromonas hydrophila is a bacteria that causes of MAS disease (motile aeromonad septicemia) in freshwater fish cultivation and can cause mass death in a fairly short period of time in some species including tilapia. There are several alternative strategies in prevention, one of which is the use of probiotic bacteria as agents for controlling or preventing this disease. One candidate for probiotics that has been molecularly identified as 16sRNA and is known to have the ability to inhibit pathogenic bacteria is B. methylotrhrophicus. The aim of this research was to study B.methylotrophicus in inhibiting A. hydrophila in Oreochromis niloticus culture. This research consisted of in vitro and in vivo test that used experiment method with completely randomized design with 4 treatments (density of 1 fishes/l) and 3 replications. The treatment consisted of a mixture of A. hydrophila 102 CFU/mL with B. methylotrophicus 109 CFU/mL (a) without addition of B. methylotrophicus (b) Addition every 3 days, (c) Addition every 5 days, (d) Addition every 7 day. 120 fishes at average weight of 17,5±1,9 g was used as experimental animals. Based on the in vitro test, the most powerful concentration of B. methylotrophicus to inhibit A. hydrophila was 109 cfu/mL with clear zone of 24,9±4,2 mm. In vivo tests show that the addition of B. methylotrophicus periodecally does not significantly affect survival rates, but can slow the gowth of A. hydrophila. Treatment D showed the highest survival rate (13.33%), followed by treatment A (6.66%), B (3.33%), and C (3.33%). These results indicate that B.methylotrophicus can prevent the gowth of A. hydrophila in vitro, and can increase SR by 6.66% in the in vivo test.

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Reprogrammed M1 macrophages with inhibited STAT3, STAT6 and/or SMAD3 extends lifespan of mice with experimental carcinoma

ZHurnal «Patologicheskaia fiziologiia i eksperimental`naia terapiia» ◽

10.25557/0031-2991.2017.02.4-9 ◽

2017 ◽

pp. 4-9

Author(s):

С.В. Калиш ◽

С.В. Лямина ◽

А.А. Раецкая ◽

И.Ю. Малышев

Keyword(s):

Tumor Growth ◽

Culture Medium ◽

Ehrlich Carcinoma ◽

Macrophage Phenotype ◽

M1 Macrophages ◽

M1 Macrophage ◽

Ec Cells ◽

Experimental Carcinoma

Цель исследования. Репрограммирование М1 фенотипа макрофагов с ингибированными факторами транскрипции М2 фенотипа STAT3, STAТ6 и SMAD и оценка их влияния на развитие карциномы Эрлиха (КЭ) in vitro и in vivo. Методика. Рост опухоли иницировали in vitro путем добавления клеток КЭ в среду культивирования RPMI-1640 и in vivo путем внутрибрюшинной инъекции клеток КЭ мышам. Результаты. Установлено, что M1макрофаги и in vitro, и in vivo оказывают выраженный противоопухолевый эффект, который превосходит антиопухолевые эффекты М1, M1, M1 макрофагов и цисплатина. Заключение. М1 макрофаги с ингибированными STAT3, STAT6 и/или SMAD3 эффективно ограничивают рост опухоли. Полученные данные обосновывают разработку новой технологии противоопухолевой клеточной терапии. Objective. Reprogramming of M1 macrophage phenotype with inhibited M2 phenotype transcription factors, such as STAT3, STAT6 and SMAD and assess their impact on the development of Ehrlich carcinoma (EC) in vitro and in vivo . Methods. Tumor growth in vitro was initiated by addition of EC cells in RPMI-1640 culture medium and in vivo by intraperitoneal of EC cell injection into mice. Results. It was found that M1 macrophages have a pronounced anti-tumor effect in vitro , and in vivo , which was greater than anti-tumor effects of M1, M1, M1 macrophages and cisplatin. Conclusion. M1 macrophages with inhibited STAT3, STAT6 and/or SMAD3 effectively restrict tumor growth. The findings justify the development of new anti-tumor cell therapy technology.

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Differentiation of mesenchymal stem cells from humans and animals into insulin-producing cells: An overview in vitro induction forms

Current Stem Cell Research & Therapy ◽

10.2174/1574888x16666201229124429 ◽

2020 ◽

Vol 16 ◽

Author(s):

Bruna O. S. Câmara ◽

Bruno M. Bertassoli ◽

Natália M. Ocarino ◽

Rogéria Serakides

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Culture Medium ◽

Adipogenic Differentiation ◽

Medium Composition ◽

Cell Therapies ◽

Insulin Producing Cells ◽

Msc Differentiation

The use of stem cells in cell therapies has shown promising results in the treatment of several diseases, including diabetes mellitus, in both humans and animals. Mesenchymal stem cells (MSCs) can be isolated from various locations, including bone marrow, adipose tissues, synovia, muscles, dental pulp, umbilical cords, and the placenta. In vitro, by manipulating the composition of the culture medium or transfection, MSCs can differentiate into several cell lineages, including insulin-producing cells (IPCs). Unlike osteogenic, chondrogenic, and adipogenic differentiation, for which the culture medium and time are similar between studies, studies involving the induction of MSC differentiation in IPCs differ greatly. This divergence is usually evident in relation to the differentiation technique used, the composition of the culture medium, the cultivation time, which can vary from a few hours to several months, and the number of steps to complete differentiation. However, although there is no “gold standard” differentiation medium composition, most prominent studies mention the use of nicotinamide, exedin-4, ß-mercaptoethanol, fibroblast growth factor b (FGFb), and glucose in the culture medium to promote the differentiation of MSCs into IPCs. Therefore, the purpose of this review is to investigate the stages of MSC differentiation into IPCs both in vivo and in vitro, as well as address differentiation techniques and molecular actions and mechanisms by which some substances, such as nicotinamide, exedin-4, ßmercaptoethanol, FGFb, and glucose, participate in the differentiation process.

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Transcriptional Profiling of Porcine Blastocysts Produced In Vitro in a Chemically Defined Culture Medium

Animals ◽

10.3390/ani11051414 ◽

2021 ◽

Vol 11 (5) ◽

pp. 1414

Author(s):

Josep M. Cambra ◽

Emilio A. Martinez ◽

Heriberto Rodriguez-Martinez ◽

Maria A. Gil ◽

Cristina Cuello

Keyword(s):

Culture Medium ◽

Embryo Quality ◽

Transcriptional Profiling ◽

Defined Medium ◽

Platelet Factor ◽

Defined Media ◽

Defined Culture ◽

The Impact

The development of chemically defined media is a growing trend in in vitro embryo production (IVP). Recently, traditional undefined culture medium with bovine serum albumin (BSA) has been successfully replaced by a chemically defined medium using substances with embryotrophic properties such as platelet factor 4 (PF4). Although the use of this medium sustains IVP, the impact of defined media on the embryonic transcriptome has not been fully elucidated. This study analyzed the transcriptome of porcine IVP blastocysts, cultured in defined (PF4 group) and undefined media (BSA group) by microarrays. In vivo-derived blastocysts (IVV group) were used as a standard of maximum embryo quality. The results showed no differentially expressed genes (DEG) between the PF4 and BSA groups. However, a total of 2780 and 2577 DEGs were detected when comparing the PF4 or the BSA group with the IVV group, respectively. Most of these genes were common in both in vitro groups (2132) and present in some enriched pathways, such as cell cycle, lysosome and/or metabolic pathways. These results show that IVP conditions strongly affect embryo transcriptome and that the defined culture medium with PF4 is a guaranteed replacement for traditional culture with BSA.

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