scholarly journals Organoid culture media containing growth factors of defined cellular activity

2018 ◽  
Author(s):  
Manuela Urbischek ◽  
Helena Rannickmae ◽  
Thomas Foets ◽  
Katharina Ravn ◽  
Marko Hyvönen ◽  
...  
Keyword(s):  
Growth Factors ◽  
Culture Media ◽  
Cost Effective ◽  
Cellular Activity ◽  
Organoid Culture ◽  
The Media ◽  
Bmp Signalling ◽  
Critical Components ◽  
Novel Applications ◽  
Gremlin 1

AbstractThe media components necessary for deriving and sustaining organoids from a number of epithelial tissues such as prostate, colon, gastric, liver, pancreas, and others have been established (1). Critical components of organoid media formulations are a set of growth factors that include EGF, R-spondins and BMP signalling antagonists such as Noggin or Gremlin. The practical limitation to organoid culture and the development of new applications for the technology is the use of defined cellular activities of growth factors in media formulations, in particular Noggin/Gremlin 1 and R-spondin 1. Here we report the production of highly pure recombinant Gremlin 1 and R-spondin 1 from bacterial expression and their use for culturing organoids. We detail the workflow for their purification, determination of cellular activity, quality control and their formulation in organoid media. The protocols we provide for generation of precisely formulated, cost-effective, organoid media of defined cellular activity will enable broader access to organoid technology and engender the development of novel applications.

scholarly journals Author Correction: Organoid culture media formulated with growth factors of defined cellular activity

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Manuela Urbischek ◽  
Helena Rannikmae ◽  
Thomas Foets ◽  
Katharina Ravn ◽  
Marko Hyvönen ◽  
...  
Keyword(s):  
Growth Factors ◽  
Culture Media ◽  
Cellular Activity ◽  
Organoid Culture

scholarly journals Organoid culture media formulated with growth factors of defined cellular activity

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Manuela Urbischek ◽  
Helena Rannikmae ◽  
Thomas Foets ◽  
Katharina Ravn ◽  
Marko Hyvönen ◽  
...  
Keyword(s):  
Growth Factors ◽  
Culture Media ◽  
Cellular Activity ◽  
Organoid Culture

Croissance et activité cellulaire du Brevibacterium casei NCDO 2049 dans du perméat de lactosérum

1997 ◽  
Vol 43 (12) ◽  
pp. 1180-1188
Author(s):  
K. M. Oulé ◽  
G. Turcotte ◽  
Y. Beaulieu
Keyword(s):  
Growth Factors ◽  
Yeast Extract ◽  
Culture Medium ◽  
Nitrogen Sources ◽  
Inhibitory Effect ◽  
Vitamin B ◽  
Cellular Activity ◽  
Whey Permeate ◽  
The Media ◽  
Influence Of Ph

Growth and cellular activity of Brevibacterium casei NCDO 2049 were studied in a whey permeate as basic culture medium. The possible inhibitory effect of the carbone substrate (undiluted or diluted permeate) on growth was investigated as well as the influence of pH of the media (controlled or not) and of the addition of nitrogen sources (organic or inorganic) or growth factors such as yeast extract or vitamin B12. Growth in undiluted permeate produced a maximal biomass (6.5 × 109 cfu/mL) that was nearly twice as much as that in diluted permeate (3.8 × 109 cfu/mL). The carbone substrate (lactose) had no inhibitory effect on growth. In undiluted permeate and an uncontrolled pH, maximal biomass was reached after 36 h of incubation, while in a pH controlled medium, twice as much time was required to obtain an equivalent biomass. In undiluted permeate and an uncontrolled pH, growth in the presence of peptone reached 22.6 × 109 cfu/mL and, in the presence of (NH4)2SO4, 12.4 × 109 cfu/mL. Adding growth factors to media with peptone resulted in the reduction of 90% of initial lactose in the presence of yeast extract and of 75% in the presence of B12 vitamin. This study indicates the possibility of reducing lactose in whey permeate when cultivating strains of the genus Brevibacterium used as maturing bacteria for certain cheese types.Key words: whey permeate, Brevibacterium casei, lactose.[Journal translation]

scholarly journals In Vitro Culture of Bovine Fibroblasts using Select Serum-Free Media Supplemented with Chlorella vulgaris Extract

2021 ◽  
Author(s):  
Galileo Defendi-Cho ◽  
Timothy Gould
Keyword(s):  
Cell Culture ◽  
Growth Factors ◽  
Cell Growth ◽  
Culture Media ◽  
Serum Free ◽  
The Media ◽  
Chlorella Algae ◽  
Free Media ◽  
Cells Cultured

Standard cell culture practices require addition of animal-derived serum to culture media to achieve adequate cell growth. Typically, 5-10% by volume of fetal bovine serum (FBS) is used, which accounts for a vast majority of the cost of media while also imposing environmental and ethical concerns associated with the use of animal serum. Here we tested the efficacy of culturing cells by replacing serum in the media with algae extract and select additives. Using LC-MS, we compared molecular signatures of FBS to Chlorella algae extracts and identified NAD(H)/NADP(H) as common and relatively abundant features in their characteristic profiles. Bovine fibroblasts, cultured in serum-free media supplemented with C. vulgaris extract and just two growth factors plus insulin, showed significant growth with enhanced viability compared to control cells cultured without serum, albeit still lower than that of controls cultured with 10% FBS. Moreover, C. vulgaris extract enhanced cell viability beyond that of cells cultured with the two growth factors and insulin alone. These results suggest that key components in serum which are essential for cell growth may also be present in C. vulgaris extract, demonstrating that it may be used at least as a partial alternative to serum for cell culture applications.

scholarly journals Borax induces osteogenesis by stimulating NaBC1 transporter via activation of BMP pathway

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Patricia Rico ◽  
Aleixandre Rodrigo-Navarro ◽  
Laura Sánchez Pérez ◽  
Manuel Salmeron-Sanchez
Keyword(s):  
Growth Factors ◽  
Cell Fate ◽  
Biomedical Applications ◽  
Cost Effective ◽  
Intrinsic Properties ◽  
Original Function ◽  
Stem Cell Fate ◽  
Osteopontin Expression ◽  
Bmp Pathway ◽  
The Media

AbstractThe intrinsic properties of mesenchymal stem cells (MSCs) make them ideal candidates for tissue engineering applications. Efforts have been made to control MSC behavior by using material systems to engineer synthetic extracellular matrices and/or include soluble factors in the media. This work proposes a simple approach based on ion transporter stimulation to determine stem cell fate that avoids the use of growth factors. Addition of borax alone, transported by the NaBC1-transporter, enhanced MSC adhesion and contractility, promoted osteogenesis and inhibited adipogenesis. Stimulated-NaBC1 promoted osteogenesis via the BMP canonical pathway (comprising Smad1/YAP nucleus translocation and osteopontin expression) through a mechanism that involves simultaneous NaBC1/BMPR1A and NaBC1/α5β1/αvβ3 co-localization. We describe an original function for NaBC1 transporter, besides controlling borate homeostasis, capable of stimulating growth factor receptors and fibronectin-binding integrins. Our results open up new biomaterial engineering approaches for biomedical applications by a cost-effective strategy that avoids the use of soluble growth factors.

Isolation, Characterization, and Differentiation of Stem Cells From Various Dental Sources: An In Vitro Study

2021 ◽  
pp. 232020682110107
Author(s):  
Sandeep S. Katti ◽  
Kishore Bhat ◽  
Chetana Bogar
Keyword(s):  
Stem Cells ◽  
Growth Factors ◽  
Statistical Analysis ◽  
Specific Growth ◽  
Culture Media ◽  
In Vitro Study ◽  
Central Research ◽  
Cell Lineages ◽  
Apical Papilla

Aim: The aim of the current study was to isolate stem cells from various dental sources such as dental pulp, periodontal ligament (PDL), and apical papilla, and to characterize stem cells by staining for the presence/absence of specific surface markers and also to differentiate stem cells into osteogenic, chondrogenic, and adipogenic cell lineages by exposing them to specific growth factors under the ideal conditions. Materials and Methods: A total of 117 samples were included in the study, consisting of 30 pulp, 50 gingival, 35 PDL, and 2 apical papilla samples. The pulp was extirpated and transported to the Central Research Laboratory. Gingival connective tissue was collected from the participants undergoing any crown lengthening procedure or any gingivectomy procedure from the Department of Periodontology. A similar procedure was also followed for apical papilla and PDL. Isolation was done followed by the identification of the cells by immunocytochemistry using different markers. Once the identity of cells was confirmed, these cells were treated with different culture media to attain 70% to 100% confluency. Then the medium was replaced with a conditioning medium containing specific growth factors for differentiation into osteogenic, chondrogenic, and adipogenic cell lineages. Result: In our study, the number of samples collected and processed was 117. The isolation rate of stem cells from the above-collected samples was 70%. Statistical analysis—no statistical analysis was done as there was no variability expected. Conclusion: Our study showed that stem cells could be isolated, differentiated, and characterized from different dental sources.

scholarly journals O80: GREMLIN-1 PROMOTES TUMOURIGENESIS AND METASTASIS IN HER2 POSITIVE BREAST CANCER

2021 ◽  
Vol 108 (Supplement_1) ◽  
Author(s):  
C Zabkiewicz ◽  
L Ye ◽  
R Hargest
Keyword(s):  
Breast Cancer ◽  
Cellular Growth ◽  
Breast Cancers ◽  
Her2 Positive ◽  
Mesenchymal Transition ◽  
Her2 Breast Cancer ◽  
Bmp Signalling ◽  
Gremlin 1

Abstract Introduction HER2 over-expression denotes poor prognosis in breast cancers.Bone morphogenetic protein(BMP) signalling is known to interact with EGF signalling, co-regulating breast cancer progression.BMP antagonist Gremlin-1 may influence breast cancer disease progression, but this remains unexplored in HER2 positive breast cancers. Method GREM1 and HER2 expression, and clinical outcomes were examined in clinical cohorts.GREM1 overexpression or pEF control plasmid were transduced into BT474 HER2+breast cancer cells. In vitro function tests using BT474 pEF and BT474GREM1cells include 2D/3D growth, migration, and expression of epithelial to mesenchymal transition(EMT)markers. Signalling cascades were examined in BT474 treated with RhGremlin-1. In vivo, BALB/c nude mice underwent either mammary injection or intra-cardiac injection of BT474pEF or BT474GREM1 cells and disease burden assessed. Result GREM1 expression correlates with HER2 in breast tumours(p=0.03) and is higher in metastatic HER2 positive cancers (p = 0.04). HER2 positive patients with high GREM1 have poor survival(p = 0.0002). BT474GREM1cells have up-regulated markers of EMT compared to control. BT474 RhGremlin-1 treated cells have active AKT pathway signalling, independent of BMP signalling. In vitro,  BT474GREM1cells significantly proliferate and migrate compared to control(p<0.05 and p < 0.001).This is confirmed in vivo,  BT474GREM1 mice grew significantly larger mammary tumours(p<0.05) and had more PETCT metastatic hotspots. Conclusion Gremlin-1 is correlated with poor outcomes in HER2 patients and promotes breast cancer cellular growth, migration and metastasis.Gremlin-1 is a novel area of research with potential as a prognostic biomarker and therapeutic target for personalised, effective, breast cancer outcomes. Take-home message BMP antagonists are gaining interest for their potential in breast cancer prognosis and therapeutics.This novel area of research shows BMP antagonist Gremlin-1 is of importance in HER2 positive breast cancers. DRAGONS DEN

scholarly journals All-Organic, Low Voltage, Transparent and Compliant Organic Field-Effect Transistor Fabricated by Means of Large-Area, Cost-Effective Techniques

2020 ◽  
Vol 10 (19) ◽  
pp. 6656
Author(s):  
Stefano Lai ◽  
Giulia Casula ◽  
Pier Carlo Ricci ◽  
Piero Cosseddu ◽  
Annalisa Bonfiglio
Keyword(s):  
Field Effect ◽  
Low Voltage ◽  
Field Effect Transistors ◽  
High Mobility ◽  
Chemical Vapor ◽  
Cost Effective ◽  
Large Area ◽  
Parylene C ◽  
Biomedical Sensing ◽  
Novel Applications

The development of electronic devices with enhanced properties of transparency and conformability is of high interest for the development of novel applications in the field of bioelectronics and biomedical sensing. Here, a fabrication process for all organic Organic Field-Effect Transistors (OFETs) by means of large-area, cost-effective techniques such as inkjet printing and chemical vapor deposition is reported. The fabricated device can operate at low voltages (as high as 4 V) with ideal electronic characteristics, including low threshold voltage, relatively high mobility and low subthreshold voltages. The employment of organic materials such as Parylene C, PEDOT:PSS and 6,13-Bis(triisopropylsilylethynyl)pentacene (TIPS pentacene) helps to obtain highly transparent transistors, with a relative transmittance exceeding 80%. Interestingly enough, the proposed process can be reliably employed for OFET fabrication over different kind of substrates, ranging from transparent, flexible but relatively thick polyethylene terephthalate (PET) substrates to transparent, 700-nm-thick, compliant Parylene C films. OFETs fabricated on such sub-micrometrical substrates maintain their functionality after being transferred onto complex surfaces, such as human skin and wearable items. To this aim, the electrical and electromechanical stability of proposed devices will be discussed.

scholarly journals The Roles of Porous Coral Sands in Initial Enrichment of Ammonia-Oxidizing Bacteria

2007 ◽  
Vol 7 ◽  
pp. 525-532
Author(s):  
Qing Guo ◽  
Zao-he Wu ◽  
Ming-liang Qian ◽  
Binhe Gu
Keyword(s):  
Culture Media ◽  
Potassium Dihydrogen Phosphate ◽  
N:P Ratio ◽  
Ammonium Nitrogen ◽  
Dihydrogen Phosphate ◽  
Ammonia Oxidizing Bacteria ◽  
Potassium Dihydrogen ◽  
Conversion Rates ◽  
Calcium Source ◽  
The Media

The purpose of this study was to investigate the roles of coral sands in the enrichment and isolation of ammonium-oxidizing bacteria (AOB). We hypothesized that the porous coral sands provided additional surface area and nutrients for the growth of periphytic AOB. In the present study, an orthogonal test was designed to compare the AOB conversion rates of ammonium-nitrogen (NH4+N) to nitrite-nitrogen (NO2--N) among various combinations of culture media. Results showed that the conversion of NH4+N to NO2--N increased significantly when the coral sands were added, implying that coral sands were beneficial to the growth of AOB. Additions of potassium dihydrogen phosphate (KH2PO4) or sodium bicarbonate (NaHCO3) to the media became unnecessary when coral sands were used, but the addition of KH2PO4was needed when the molar nitrogen to phosphorus (N:P) ratio reached 10 in the enrichment media using calcium carbonate (CaCO3) powder as a calcium source.

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