Foetal bovine serum influence on in vitro extracellular vesicle analyses

Present study was carried out to find the effect of combining EGF with IGF, cysteine and sodium pyruvate singly as additive in a medium consisting of TCM-199 + 100 µl/ml foetal bovine serum + 100 µM/ml cysteamine + 1 µg/ml 17â- Oestradiol + 5 µg/ml pFSH + 5µg/ml oLH + 10 per cent follicular fluid and 10 per cent oestrous goat serum on in-vitro maturation (IVM) of caprine oocytes on incubation at 38.50C for 24 hours in a CO2 incubator maintaining 5 per cent CO2 under humidified condition. The additives comprised 10 ng/ml EGF + 50 ng/ml IGF-1, 10 ng/ml EGF + 600 µM/ml cysteine and 10 ng/ ml EGF + 0.2 mM/ml sodium pyruvate. The IVM rate of oocytes on the basis of cumulus cells expansion and nuclear maturation was found to be significantly higher (P less than 0.05) with EGF + IGF-1 (88.74 ± 1.85% and 61.71 ± 1.61%) than with EGF + sodium pyruvate (82.86 ± 0.97% and 54.62 ± 1.88%), EGF + cysteine ( 78.58 ± 1.45% and 49.02 ± 1.52%) and without additive (control) (75.27 ± 1.58% and 43.03 ± 1.48%).

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The effect of foetal bovine serum supplementation upon the lactate dehydrogenase cytotoxicity assay: Important considerations for in vitro toxicity analysis

Toxicology in Vitro ◽

10.1016/j.tiv.2015.10.007 ◽

2015 ◽

Vol 30 (1) ◽

pp. 300-308 ◽

Cited By ~ 18

Author(s):

Martin G. Thomas ◽

Roxanne M. Marwood ◽

Anna E. Parsons ◽

Richard B. Parsons

Keyword(s):

Lactate Dehydrogenase ◽

Bovine Serum ◽

Cytotoxicity Assay ◽

In Vitro Toxicity ◽

Serum Supplementation ◽

Toxicity Analysis ◽

Foetal Bovine Serum

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The influence of myo-inositol on phosphatidylglycerol synthesis by rat type II pneumonocytes

Biochemical Journal ◽

10.1042/bj2120811 ◽

1983 ◽

Vol 212 (3) ◽

pp. 811-818 ◽

Cited By ~ 19

Author(s):

J E Bleasdale ◽

N E Tyler ◽

F N Busch ◽

J G Quirk

Keyword(s):

Incubation Medium ◽

Bovine Serum ◽

Total Lipid Extract ◽

Type Ii ◽

Rat Serum ◽

Adult Rat ◽

Foetal Rat ◽

Inositol Uptake ◽

Foetal Bovine Serum

Type II pneumonocytes isolated from adult rat lung were incubated in a serum-free medium containing [14C]glycerol and the incorporation of 14C into glycerophospholipids was measured. After 24 h, more than 80% of the 14C incorporated into total lipids or into phosphatidylcholine and approx. 90% of the 14C incorporated into phosphatidylglycerol after 24 h was recovered in the glycerophosphoester moieties of these molecules. Supplementation of the incubation medium with foetal-bovine serum (10%, v/v) did not alter the incorporation of [14C]glycerol by type II pneumonocytes after 24 h into either a total lipid extract or phosphatidylcholine. In the presence of foetal-bovine serum, however, the incorporation of 14C into phosphatidylglycerol was decreased and the incorporation of 14C into phosphatidylinositol was increased. In the absence of foetal-bovine serum, the incorporation of 14C into phosphatidylglycerol was decreased progressively as the concentration of myo-inositol in the incubation medium was increased. The range of concentration (0.04-0.50 mM) over which myo-inositol had the greatest influence on [14C]glycerol incorporation into phosphatidylglycerol by type II pneumonocytes in vitro encompassed the concentration range measured in foetal-rat serum late in gestation. At 4 days before birth, the concentration of myo-inositol in foetal-rat serum was 0.36 mM and decreased to 0.23 mM 1 day before birth. The concentration of myo-inositol in adult rat serum increased from 0.03 mM to 0.06 mM during pregnancy. Isolated rat type II pneumonocytes were found to take up myo-inositol by a saturable process. A half-maximal rate of myo-inositol uptake occurred at a concentration of myo-inositol of 0.29 mM. The results of this investigation are consistent with the hypothesis that late in gestation there is a decreasing availability of myo-inositol to the foetal lungs and that this favours the biosynthesis of phosphatidylglycerol for surfactant at the expense of phosphatidylinositol biosynthesis.

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Adaptation of human diploid fibroblasts in vitro to serum from different sources

Journal of Cell Science ◽

10.1242/jcs.61.1.289 ◽

1983 ◽

Vol 61 (1) ◽

pp. 289-297 ◽

Cited By ~ 1

Author(s):

G.B. Zamansky ◽

C. Arundel ◽

H. Nagasawa ◽

J.B. Little

Keyword(s):

Bovine Serum ◽

Horse Serum ◽

Fibroblast Cell ◽

Human Diploid Fibroblasts ◽

Generation Times ◽

Different Sources ◽

Cell Volumes ◽

Foetal Bovine Serum ◽

Fibroblast Cell Lines

The growth of two human diploid skin fibroblast cell lines, originally grown in medium supplemented with foetal bovine serum and later adapted to medium supplemented with newborn bovine, bovine calf or horse serum, has been studied. Prolonged generation times increased cell volumes and decreased plating efficiencies were observed in cultures grown in newborn bovine, bovine calf or horse serum. In general, the deleterious effects were most severe as a result of growth in bovine calf or horse serum. In the light of the present findings, we believe investigators should exert great caution in switching human fibroblast cultures from foetal bovine serum to alternative sera, even at times of scarcity and high prices.

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Kultur Sel Baby Hamster Kidney (BHK) Menggunakan Media Dulbecco's Modified Eagle Medium (DMEM)

BIOTROPIC The Journal of Tropical Biology ◽

10.29080/biotropic.2017.1.1.1-8 ◽

2017 ◽

Vol 1 (1) ◽

pp. 1-8

Author(s):

Mashita Andiana

Keyword(s):

Cell Line ◽

Bovine Serum ◽

Calf Serum ◽

Baby Hamster Kidney ◽

Counting Chamber ◽

Foetal Bovine Serum

Kultur sel merupakan suatu proses saat sel hidup ditempatkan ke dalam suatu media yang dapat membuat sel tersebut berkembang biak atau tumbuh secara in vitro, Kultur sel dapat berupa kultur sel primer maupun cell line, Metode dalam kultur sel terdiri atas kultur monolayer dan kultur suspensi. Pembuatan media kultur untuk pertumbuhan sel diusahakan memenuhi kriteria. Konstituen dasar dari media kultur yang paling banyak digunakan adalah BSS (Balanced Sald Solution). Untuk mendapatkan pertumbuhan sel yang optimal, media kultur ditambahkan serum. Serum yang biasa digunakan dalam kultur adalah serum anak sapi (calf serum), serum fetus sapi (foetal bovine serum), serum kuda dan serum manusia. Perhitungan sel menggunakan counting chamber. Metode yang digunakan adalah revival kemudian split sel, stor sel dan menghitung sel. Hasil revival jumlah sel tertinggi didapat dari botol nomer 3 dengan jumlah sel 310 yaitu botol dengan sel BHK label tahun 2016, dan hasil split sel yang paling tinggi juga didapat dari botol kultur nomer 3 dengan jumlah sel 345 dan 311.

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Measuring the density and viscosity of culture media for optimized computational fluid dynamics analysis of in vitro devices

10.1101/2020.08.25.266221 ◽

2020 ◽

Author(s):

Christine Poon

Keyword(s):

Fluid Dynamics ◽

Computational Fluid Dynamics ◽

Cell Lines ◽

Dynamic Viscosity ◽

Bovine Serum ◽

Culture Medium ◽

Cfd Analysis ◽

Culture Systems ◽

Foetal Bovine Serum

AbstractCulture medium is frequently modelled as water in computational fluid dynamics (CFD) analysis of in vitro culture systems involving flow, such as bioreactors and organ-on-chips. However, culture medium can be expected to have different properties to water due to its higher solute content. Furthermore, cellular activities such as metabolism and secretion of ECM proteins alter the composition of culture medium and therefore its properties during culture. As these properties directly determine the hydromechanical stimuli exerted on cells in vitro, these, along with any changes during culture must be known for CFD modelling accuracy and meaningful interpretation of cellular responses. In this study, the density and dynamic viscosity of DMEM and RPMI-1640 media supplemented with typical concentrations of foetal bovine serum (0, 5, 10 and 20% v/v) were measured to serve as a reference for computational design analysis. Any changes in the properties of medium during culture were also investigated with NCI-H460 and HN6 cell lines. The density and dynamic viscosity of the media increased proportional to the % volume of added foetal bovine serum (FBS). Importantly, the viscosity of 5% FBS-supplemented RPMI-1640 was found to increase significantly after 3 days of culture of NCI-H460 and HN6 cell lines, with distinct differences between magnitude of change for each cell line. Finally, these experimentally-derived values were applied in CFD analysis of a simple microfluidic device, which demonstrated clear differences in maximum wall shear stress and pressure between fluid models. Overall, these results highlight the importance of characterizing model-specific properties for CFD design analysis of cell culture systems.

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Feasibility of Human Platelet Lysate as an Alternative to Foetal Bovine Serum for In Vitro Expansion of Chondrocytes

International Journal of Molecular Sciences ◽

10.3390/ijms22031269 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1269

Author(s):

Ling Ling Liau ◽

Muhammad Najib Fathi bin Hassan ◽

Yee Loong Tang ◽

Min Hwei Ng ◽

Jia Xian Law

Keyword(s):

Bovine Serum ◽

Human Platelet ◽

Pathogen Transmission ◽

Platelet Lysate ◽

Joint Disease ◽

Suitable Alternative ◽

In Vitro Expansion ◽

Human Platelet Lysate ◽

Foetal Bovine Serum

Osteoarthritis (OA) is a degenerative joint disease that affects a lot of people worldwide. Current treatment for OA mainly focuses on halting or slowing down the disease progress and to improve the patient’s quality of life and functionality. Autologous chondrocyte implantation (ACI) is a new treatment modality with the potential to promote regeneration of worn cartilage. Traditionally, foetal bovine serum (FBS) is used to expand the chondrocytes. However, the use of FBS is not ideal for the expansion of cells mean for clinical applications as it possesses the risk of animal pathogen transmission and animal protein transfer to host. Human platelet lysate (HPL) appears to be a suitable alternative to FBS as it is rich in biological factors that enhance cell proliferation. Thus far, HPL has been found to be superior in promoting chondrocyte proliferation compared to FBS. However, both HPL and FBS cannot prevent chondrocyte dedifferentiation. Discrepant results have been reported for the maintenance of chondrocyte redifferentiation potential by HPL. These differences are likely due to the diversity in the HPL preparation methods. In the future, more studies on HPL need to be performed to develop a standardized technique which is capable of producing HPL that can maintain the chondrocyte redifferentiation potential reproducibly. This review discusses the in vitro expansion of chondrocytes with FBS and HPL, focusing on its capability to promote the proliferation and maintain the chondrogenic characteristics of chondrocytes.

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695 Oral delivery of a microbial extracellular vesicle induces potent anti-tumor immunity in mice

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-sitc2020.0695 ◽

2020 ◽

Vol 8 (Suppl 3) ◽

pp. A737-A737

Author(s):

Loise Francisco-Anderson ◽

Mary Abdou ◽

Michael Goldberg ◽

Erin Troy ◽

...

Keyword(s):

Tumor Microenvironment ◽

Tumor Growth ◽

Extracellular Vesicles ◽

Tumor Immunity ◽

Extracellular Vesicle ◽

The Body ◽

Checkpoint Inhibition ◽

Small Intestinal ◽

The Impact

BackgroundThe small intestinal axis (SINTAX) is a network of anatomic and functional connections between the small intestine and the rest of the body. It acts as an immunosurveillance system, integrating signals from the environment that affect physiological processes throughout the body. The impact of events in the gut in the control of tumor immunity is beginning to be appreciated. We have previously shown that an orally delivered single strain of commensal bacteria induces anti-tumor immunity preclinically via pattern recognition receptor-mediated activation of innate and adaptive immunity. Some bacteria produce extracellular vesicles (EVs) that share molecular content with the parent bacterium in a particle that is roughly 1/1000th the volume in a non-replicating form. We report here an orally-delivered and gut-restricted bacterial EV which potently attenuates tumor growth to a greater extent than whole bacteria or checkpoint inhibition.MethodsEDP1908 is a preparation of extracellular vesicles produced by a gram-stain negative strain of bacterium of the Oscillospiraceae family isolated from a human donor. EDP1908 was selected for its immunostimulatory profile in a screen of EVs from a range of distinct microbial strains. Its mechanism of action was determined by ex vivo analysis of the tumor microenvironment (TME) and by in vitro functional studies with murine and human cells.ResultsOral treatment of tumor-bearing mice with EDP1908 shows superior control of tumor growth compared to checkpoint inhibition (anti-PD-1) or an intact microbe. EDP1908 significantly increased the percentage of IFNγ and TNF producing CD8+ CTLs, NK cells, NKT cells and CD4+ cells in the tumor microenvironment (TME). EDP1908 also increased tumor-infiltrating dendritic cells (DC1 and DC2). Analysis of cytokines in the TME showed significant increases in IP-10 and IFNg production in mice treated with EDP1908, creating an environment conducive to the recruitment and activation of anti-tumor lymphocytes.ConclusionsThis is the first report of striking anti-tumor effects of an orally delivered microbial extracellular vesicle. These data point to oral EVs as a new class of immunotherapeutic drugs. They are particularly effective at harnessing the biology of the small intestinal axis, acting locally on host cells in the gut to control distal immune responses within the TME. EDP1908 is in preclinical development for the treatment of cancer.Ethics ApprovalPreclinical murine studies were conducted under the approval of the Avastus Preclinical Services’ Ethics Board. Human in vitro samples were attained by approval of the IntegReview Ethics Board; informed consent was obtained from all subjects.

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A Precipitin-like Reaction of the Hemolymph of the Lobster Homarus americanus

Journal of the Fisheries Research Board of Canada ◽

10.1139/f69-127 ◽

1969 ◽

Vol 26 (5) ◽

pp. 1392-1397 ◽

Cited By ~ 10

Author(s):

James E. Stewart ◽

Diane M. Foley

Keyword(s):

Bovine Serum Albumin ◽

Serum Albumin ◽

Bovine Serum ◽

Serum Proteins ◽

Homarus Americanus ◽

Test Period ◽

Fluorescent Material

The levels of fluorescent material in the hemolymph of lobsters injected with serum proteins from lobster hemolymph labelled with fluorescein remained relatively constant over a 6-day test period; the levels in lobsters injected with bovine serum albumin labelled with fluorescein declined rapidly. A precipitin-like reaction was observed when lobster hemolymph serum was titrated with bovine serum albumin in vitro.

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