Development in culture of rat foetuses explanted at 12·5 and 13·5 days of gestation

Development ◽  
1973 ◽  
Vol 29 (2) ◽  
pp. 473-483
Author(s):  
D. L. Cockroft
Keyword(s):  
Protein Content ◽  
Culture Medium ◽  
Yolk Sac ◽  
Gas Mixture ◽  
Rat Serum ◽  
Simple Operation ◽  
Flowing Medium ◽  
Economic Culture ◽  
Capillary Circulation ◽  
Somite Number

1. As a result of the relatively simple operation of opening the yolk sac and thus exposing the foetal capillary circulation to flowing medium, it has proved possible to grow 12·5- and 13·5-day rat foetuses for a period of 42 h in culture. 2. 25% rat serum, 75% Tyrode has been found to be a satisfactory and economic culture medium, and has the added attraction that foetal survival is improved compared with that obtained in whole rat serum. 3. For both 12·5- and 13·5-day foetuses grown with open yolk sacs, a gas mixture of 95% O2, 5% CO2 in equilibrium at 1 atm with culture medium flowing at about 15 ml/min has been found to give the best results. 4. Under these conditions, foetuses explanted at 12·5 days increased their protein content from 1·0–1·3 mg to 2·3–2·7 mg during culture. Their somite number was 40–44 at explantation and reached 50–55 during culture. 5. Foetuses explanted at 13·5 days increased their protein content from 3·2–4·0 mg to 6·1–7·5 mg during culture. Their somite number was 51–55 at explantation and reached 60–63 during culture.

scholarly journals Inhibition of proteolysis in rat yolk sac as a cause of teratogenesis. Effects of leupeptin in vitro and in vivo

Development ◽  
1983 ◽  
Vol 78 (1) ◽  
pp. 183-193
Author(s):  
Stuart J. Freeman ◽  
John B. Lloyd
Keyword(s):  
Amino Acids ◽  
Protein Content ◽  
Culture Medium ◽  
Specific Inhibitor ◽  
Yolk Sac ◽  
Cysteine Proteinases ◽  
Rat Serum ◽  
Pregnant Rats ◽  
Stage Of Development

Conceptuses from 9·5-day pregnant rats were cultured for 48 h in heat-inactivated homologous serum to which leupeptin, a specific inhibitor of the lysosomal cysteine-proteinases, was added for the final or the penultimate 6h. The presence of leupeptin (25 µg/ml or above) increased the protein content of yolk sacs at harvesting to approximately twice the control value. The protein content of the embryo at harvesting was lower than that of controls. When 125I-labelled polyvinylpyrrolidone was added to the culture serum for the final 6 h of culture, radioactivity was found in the yolk sac at harvesting, but not in the embryo. The presence of leupeptin did not affect the rate of uptake of the radiolabelled macromolecule by the yolk sac, nor facilitate its entry into the embryo. When formaldehyde-denatured 125I-labelled bovine serum albumin was added to the culture medium for the final 6 h of culture, little radioactivity was found in the yolk sac at harvesting, and barely any was found in the embryo. Trichloroacetic acid-soluble radioactivity was found in the culture serum. The presence of leupeptin sharply increased the levels of radioactivity in the yolk sac (but not the embryo) and sharply decreased the acid-soluble radioactivity of the culture medium. When rat serum whose proteins were labelled with [3H]leucine was used as culture medium, radioactivity was found in both yolk sac and embryo at harvesting. The presence of leupeptin increased the amount found in the yolk sac and decreased that found in the embryo. These results are interpreted as follows. Leupeptin enters the lysosomes of the yolk sac, inhibiting their cysteine proteinases. The digestion of proteins pinocytosed by the yolk sac is consequently inhibited, resulting in the accumulation of protein by the yolk sac and a decreased flow of amino acids to the embryo. Leupeptin (50 mg/kg), injected into pregnant rats at either 8·5 days or 9·5 days of gestation, induced congenital malformation in the offspring. It is proposed that leupeptin exerts its teratogenic action by inhibiting proteolysis in the lysosomes of the yolk sac, and so depriving the developing embryo of its supply of amino acids at a critical stage of development.

Characterization of protein production by caprine placental membranes: identification and immunolocalization of retinol-binding protein

1995 ◽  
Vol 146 (3) ◽  
pp. 527-534 ◽  
Author(s):  
K H Liu ◽  
J C Huang ◽  
J D Godkin
Keyword(s):  
Protein Production ◽  
Culture Medium ◽  
De Novo ◽  
Binding Protein ◽  
Yolk Sac ◽  
Retinol Binding Protein ◽  
Minimum Essential Medium ◽  
Placental Membranes ◽  
Extraembryonic Membranes

Abstract Caprine chorion, allantois and amnion from days 23, 28, 35, 39 and 45, and yolk sac from day 23 of pregnancy were isolated by dissection and cultured for 24 h in modified minimum essential medium in the presence of [35S] methionine. De novo-synthesized proteins released into the culture medium were analyzed by two-dimensional PAGE and fluorography. Patterns of protein production by these isolated extraembryonic membranes remained relatively unchanged from days 23 to 45 of pregnancy. Electrophoretic profiles of proteins synthesized by allantois and amnion were identical but distinct from that produced by chorion. Yolk sac was the major source of serum-like proteins. An acidic (pI 5·3–6·3) 22 kDa protein, which consisted of four isoelectric variants, was produced by all extraembryonic membranes and demonstrated to immunoreact with antiserum produced against bovine placental retinol-binding protein (RBP). Limited N-terminal sequence analysis of one major isoform indicated that the protein had complete homology with bovine RBP over the first 15 amino acids. Immunoreactive RBP was localized in epithelial cells lining the chorion, allantois and amnion. In this study, we have characterized and compared protein production by isolated extraembryonic membranes through days 23 to 45 of pregnancy and identified the 22 kDa protein as caprine RBP of placental origin. Journal of Endocrinology (1995) 146, 527–534

Effect of yolk-sac antibody on rat embryos grown in culture

Development ◽  
1972 ◽  
Vol 27 (3) ◽  
pp. 543-553
Author(s):  
D. A. T. New ◽  
R. L. Brent
Keyword(s):  
Direct Contact ◽  
Culture Medium ◽  
Gamma Globulin ◽  
Yolk Sac ◽  
Amniotic Cavity ◽  
Pregnant Rats ◽  
Rat Embryos ◽  
Visceral Yolk Sac ◽  
Primary Action

Rat embryos, explanted with their embryonic membranes during the early stages of organogenesis ( days gestation), were grown in culture in roller tubes. Yolk-sac antibody (sheep anti rat yolk-sac gamma globulin), known to be teratogenic when injected into pregnant rats, was added to the culture medium. At concentrations of 0·1 mg/ml or more the antibody caused gross retardation of growth and differentiation. Injection of antibody into the amniotic cavity so that it had direct contact with the embryo, or between the amnion and yolk sac so that it was in contact with the mesodermal surface of the yolk sac, had little or no effect on development of the embryo or its membranes. These in vitro experiments indicate that yolk-sac antibody has an effect on development independent of any immunological reaction of the mother, and the primary action is probably on the visceral yolk-sac endoderm.

GATA-1 Protein Levels Are Not Altered in CD34+ and CD61+ Cells, but NF-κb Protein Levels Are Elevated in Patients with Idiopathic Myelofibrosis

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4498-4498
Author(s):  
Jen-Chin Wang ◽  
Seema Naik ◽  
Chi Chen ◽  
Ying-Yi Xiao ◽  
Tsong H. Chang
Keyword(s):  
Protein Content ◽  
Culture Medium ◽  
Myeloproliferative Disorders ◽  
Idiopathic Myelofibrosis ◽  
Hematopoietic Progenitor ◽  
Protein Levels ◽  
Nuclear Extracts ◽  
Cd34 Cells ◽  
Stem Cell Growth Factor ◽  
Cells Cultured

Abstract GATA-1 and NF-kB are important in megakaryocytopoiesis and pathogenesis of idiopathic myelofibrosis (IMF). Previous studies have shown that GATA-1 levels were normal in mRNA but were lower in protein content in CD61+ cells of patients with IMF, and we reported previously that GATA-1 protein levels were not altered in CD61+ cells. The current study includes more patients and also measured NF-kB levels. ELISA assay (GATA-1 and NF-kB ELISA kit, Panomics Inc. CA) was used to determine and compare the protein content of nuclear extracts of blood hematopoietic progenitor cells (CD34+) and megakaryocytes (CD61+ cells) from patients with IMF, patients with other myeloproliferative disorders, and with controls. CD61+ cells were obtained from 10-day cultures of blood CD34+ cells in serum-free culture medium with thrombopoietin and stem cell growth factor. Our study confirmed that GATA-1 protein levels are not altered in megakaryocytes (cultured CD 61+ cells) in patients with IMF (Fig. 1) and NF-kB levels are increased (Table 1). We conclude that GATA-1 protein levels are not altered in patients with IMF compared with controls. Fig 1. GATA-1 Protein levels in CD61+ cells Cultured from Patients with IMF & Others Fig 1. GATA-1 Protein levels in CD61+ cells Cultured from Patients with IMF & Others Table 1. NF-kB Levels (ng/2μg protein) IMF Controls CD34+ cells 1.39 ± 0.27 1.10 ± 0.24 CD61+ cells 1.54 ± 0.52 0.45 ± 0.22

scholarly journals Environmental conditions modulate the protein content and immunomodulatory activity of extracellular vesicles produced by the probiotic Propionibacterium freudenreichii

2020 ◽  
Author(s):  
Vinícius de Rezende Rodovalho ◽  
Brenda Silva Rosa da Luz ◽  
Aurélie Nicolas ◽  
Fillipe Luiz Rosa do Carmo ◽  
Julien Jardin ◽  
...  
Keyword(s):  
Protein Content ◽  
Extracellular Vesicles ◽  
Environmental Conditions ◽  
Culture Medium ◽  
Surface Proteins ◽  
Bioactive Molecules ◽  
Propionibacterium Freudenreichii ◽  
Therapeutic Applications ◽  
Immunomodulatory Properties

Propionibacterium freudenreichii is a probiotic Gram-positive bacterium with promising immunomodulatory properties. It modulates regulatory cytokines, mitigates the inflammatory response in vitro and in vivo. These properties were initially attributed to specific bacterial surface proteins. Recently, we showed that extracellular vesicles (EVs) produced by P. freudenreichii CIRM-BIA129 mimic the immunomodulatory features of parent cells in vitro (i.e. modulating NF-κB transcription factor activity and IL-8 release) which underlies the role of EVs as mediators of the probiotic effects of the bacterium. The modulation of EV properties, and particularly of those with potential therapeutic applications such as the EVs produced by the probiotic P. freudenreichii, is one of the challenges in the field to achieve efficient yields with the desired optimal functionality. Here we evaluated whether the culture medium in which the bacteria are grown could be used as a lever to modulate the protein content and hence the properties of P. freudenreichii CIRM-BIA129 EVs. The physical, biochemical and functional properties of EVs produced from cells cultivated on laboratory Yeast Extract Lactate (YEL) medium and cow milk ultrafiltrate (UF) medium were compared. UF-derived EVs were more abundant, smaller in diameter and displayed more intense anti-inflammatory activity than YEL-derived EVs. Furthermore, the growth media modulated EV content in terms of both the identities and abundances of their protein cargos, suggesting different patterns of interaction with the host. Proteins involved in amino acid metabolism and central carbon metabolism were modulated, as were the key surface proteins mediating host-propionibacteria interactions. Importance Extracellular vesicles (EVs) are cellular membrane-derived nanosized particles that are produced by most cells in all three kingdoms of life. They play a pivotal role in cell-cell communication through their ability to transport bioactive molecules from donor to recipient cells. Bacterial EVs are important factors in host-microbe interactions. Recently we have shown that EVs produced by the probiotic P. freudenreichii exhibited immunomodulatory properties. We evaluate here the impact of environmental conditions, notably culture media, on P. freudenreichii EV production and function. We show that EVs display considerable differences in protein cargo and immunomodulation depending on the culture medium used. This work offers new perspectives for the development of probiotic EV-based molecular delivery systems, and reinforces the optimization of growth conditions as a tool to modulate the potential therapeutic applications of EVs.

scholarly journals Kandungan Protein Spirulina platensis Pada Media Kultur Dengan Konsentrasi Nitrat (KNO3 ) Yang Berbeda

2018 ◽  
Vol 7 (2) ◽  
pp. 98
Author(s):  
Saniyatul Ulya ◽  
Sri Sedjati ◽  
Ervia Yudiati
Keyword(s):  
Protein Content ◽  
Spirulina Platensis ◽  
Culture Medium ◽  
Culture Media ◽  
Nitrate Concentration ◽  
Block Design ◽  
Quality Measurement ◽  
Daily Basis ◽  
Natural Food ◽  
Protein Levels

 Spirulina platensis merupakan mikroalga hijau biru yang mengandung nutrisi protein tinggi sehingga banyak digunakan sebagai pakan alami. Pertumbuhan dan kandungan protein mikroalga dipengaruhi oleh beberapa faktor, salah satunya adalah pemberian makronutrien pada media kultur mikroalga. Penelitian ini bertujuan untuk melihat pengaruh pertumbuhan dan kandungan protein pada mikroalga S. platensis dengan pemberian konsentrasi nitrat yang berbeda. Rancangan penelitian yang digunakan adalah rancangan acak blok dengan tiga kali pengulangan. Perlakuan konsentrasi nitrat berbeda yang diberikan adalah 50 ppm, 100 ppm, dan 150 ppm. Perhitungan kepadatan dan pengukuran parameter kualitas air dilakukan setiap hari. Pemanenan dilakukan pada hari ke – empat. Kadar prorein dianalisis dengan menggunakan metode Kjedahl. Hasil penelitian pertumbuhan S. platensis menunjukkan nilai kepadatan sel S. platensis tertinggi pada hari ke – empat berada pada perlakuan C dengan konsentrasi nitrat 150 ppm (169,58 . 103 sel/mm3). Hasil analisis ANOVA menunjukkan bahwa konsentrasi nitrat berpengaruh terhadap pertumbuhan S. platensis (p < 0,05) namun perbedaan konsentrasi nitrat tidak berpengaruh pada kadar protein (p ≥ 0,05). Berdasarkan hasil penelitian dapat disimpulkan bahwa konsentrasi nitrat yang ditambahkan pada media kultur S. platensis berpengaruh terhadap pertumbuhan S. platensis namun tidak berpengaruh pada kandungan proteinnya.  Protein Content of Spirulina platensis in Different Culture Media with Nitrate (KNO3) Concentration Spirulina platensis is green-blue microalgae that contain high protein nutrient and could be used as natural food. Growth and protein content of microalgae are influenced by several factors and one of those is giving macronutrient to microalgae’s culture medium. The purpose of this research is to compare the growth and protein content of the S. platensis with different nitrate consentrations.The research design used was a completely randomized block design with three repetitions. The different nitrate concentration treatments were 50 ppm, 100 ppm and 150 ppm. Determination of density and water quality measurement parameters was done on daily basis. Spirulina platensis was harvested done on fourth day of culture. Protein levels were analyzed by Kjedahl method. The result of the S. platensis growth that the highest density on day fourth in C treatment with 150 ppm nitrate consentration (169,58 . 103 sel/mm3). The result of ANOVA analysis show that the concentration of nitrate affected on S. platensis growth (p < 0,05) but the difference of nitrate concentration wasn’t affected in protein analysis (p ≥ 0,05). Based on the result of this research, it can be concluded that the concentration of nitrate added to the S. platensis culture medium effectively and improved the growth of S. platensis but had no effect on the protein content. 

scholarly journals Cultivation of Chlorella vulgaris in Sequential Flow Photobioreactor System: Influence of Recycled Culture Medium on Growth, Lipid and Protein Content

2021 ◽  
Vol 721 (1) ◽  
pp. 012013
Author(s):  
Yaleeni Kanna Dasan ◽  
Man Kee Lam ◽  
Suzana Yusup ◽  
Jun Wei Lim ◽  
Keat Teong Lee ◽  
...  
Keyword(s):  
Protein Content ◽  
Chlorella Vulgaris ◽  
Culture Medium

The role of the visceral yolk sac in mediating protein utilization by rat embryos cultured in vitro

Development ◽  
1981 ◽  
Vol 66 (1) ◽  
pp. 223-234
Author(s):  
Stuart J. Freeman ◽  
Felix Beck ◽  
John B. Lloyd
Keyword(s):  
Serum Proteins ◽  
Yolk Sac ◽  
Rat Serum ◽  
Pregnant Rats ◽  
Digestion Product ◽  
Background Levels ◽  
Visceral Yolk Sac ◽  
Chorioallantoic Placenta

Conceptuses from 9·5-day pregnant rats have been cultured for 48 h in heat-inactivated homologous serum. Embryonic development was normal. The protein contents of embryos and visceral yolk sacs after different periods of culture were recorded. When 125-labelled polyvinylpyrrolidone or [3H]dextran were added to the culture serum, radioactivity was accumulated by the yolk sac, but only background levels were detected in the embryo itself. The amount of radioactivity found in the yolk sac varied with the length of the interval before harvesting during which 125 I-labelled PVP or [3H]dextran was present. When formaldehyde-denatured 125 I-labelled bovine serum albumin was added to the culture serum, little radioactivity accumulated in the yolk sac and only background levels were found in the embryo. Trichloroacetic acid-soluble radioactivity steadily appeared in the culture serum, however. When conceptuses were cultured in glucose- and vitamin-supplemented dialysed serum from rats injected 2 h previously with [3H]leucine, radioactivity was found in both embryos and yolk sacs. The amount of radioactivity in these tissues increased with duration of exposure to 3H-labelled serum proteins. After short exposures little of the yolk sac and embryonic radioactivity was acid-insoluble, but this proportion increased with duration of exposure. These results are interpreted as follows. Intact macromolecules cannot enter the cells of the embryo itself, but are captured by pinocytosis into the cells of the visceral yolk-sac endoderm. Indigestible macromolecules such as 125 I-labelled polyvinylpyrrolidone and [3H]- dextran accumulate within the yolk-sac lysosomes, but proteins are digested there by the lysosomal enzymes. The radiolabelled digestion product of 125 I-labelled bovine albumin is [125 I]iodotyrosine, which cells cannot utilize and so is excreted into the culture serum. The labelled digestion product of the 3H-labelled rat serum proteins is [3H]leucine, which is used for protein synthesis in both embryo and yolk sac. The experiments provide direct evidence for the long-suspected role of the yolk sac in mediating embryonic nutrition in the period of development prior to the establishment of a functional chorioallantoic placenta.

Toxicity of lead on microorganisms: Interaction with protein content of culture medium

1987 ◽  
Vol 2 (2) ◽  
pp. 175-191
Author(s):  
J. Santos Oliveira ◽  
A. Rodrigues ◽  
B. Mendes ◽  
M. Dias
Keyword(s):  
Protein Content ◽  
Culture Medium
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