The Microenvironment in an Experimental Model of Acute Pancreatitis Can Modify the Formation of the Protein Corona of sEVs, with Implications on Their Biological Function DOI Open Access
Olga Armengol-Badia, Jaxaira Maggi, Carmen Casal

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(23), P. 12969 - 12969

Published: Dec. 2, 2024

A considerable number of the physiological functions extracellular vesicles are conditioned by protein corona attached to their surface. The composition this is initially defined during intracellular synthesis, but it can be subsequently modified interactions with microenvironment. Here, we evaluated how small exposed inflammatory environment generated in acute pancreatitis and what functional changes occur as a result these modifications. Small obtained from pancreatic cell line were incubated ascitic fluid experimental rats. Using proteomic techniques, detected appearance new proteins an increase uptake certain types response induced cells. inhibition different pattern recognition receptors reversed activation, indicating that some effects could due binding damage-associated molecular patterns corona. All indicates pathologies such pancreatitis, characterized intense tissue damage, microenvironment substantially influences vesicles, thus altering behavior enhancing activity.

Language: Английский

Heat inactivation of foetal bovine serum performed after EV‐depletion influences the proteome of cell‐derived extracellular vesicles DOI Creative Commons
Ornella Urzì,

Markus Bergqvist,

Cecilia Lässer

et al.

Journal of Extracellular Vesicles, Journal Year: 2024, Volume and Issue: 13(1)

Published: Jan. 1, 2024

The release of extracellular vesicles (EVs) in cell cultures as well their molecular cargo can be influenced by culture conditions such the presence foetal bovine serum (FBS). Although several studies have evaluated effect removing FBS-derived EVs ultracentrifugation (UC), less is known about influence FBS heat inactivation (HI) on cell-derived EVs. To assess this, three protocols based different combinations EV depletion UC and HI were evaluated, including ultracentrifuged but not inactivated (no-HI FBS), before (HI-before EV-depl after (HI-after FBS). We isolated large (L-EVs) small (S-EVs) from treated ways, we found that S-EV pellet HI-after was larger than no-HI HI-before FBS. Transmission electron microscopy, protein quantification, particle number evaluation showed significantly increased amount S-EVs had no significant L-EVs. Consequently, quantity lines cultured media supplemented with increased. Quantitative mass spectrometry analysis content when compared to Moreover, show quantified proteins could ascribed human origin, thus demonstrating mistakenly attributed conclude performed results changes proteome, molecules co-isolate contaminate used subsequent cultures. Our recommendation is, therefore, always perform prior depletion.

Language: Английский

Citations

4
The effect of culture media and seeding densities on the human malignant testicular germ cell line (NT2/D1) growth pattern DOI Creative Commons

Rawabi S. Altuwayjiri,

Ibtesam S. Almami

Journal of Biological Research - Bollettino della Società Italiana di Biologia Sperimentale, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 5, 2025

Testicular cancer is a common malignancy in young males, comprising 5% of urological cancers and 1% all male cancers. The NT2/D1 cell line, pluripotent embryonal carcinoma, valuable developmental biology research due to its differentiation potential into neurons glial cells. Although Fetal Bovine Serum (FBS) often used promote growth, limitations necessitate optimized culture conditions. Monitoring growth health through methods such as counting, viability assays, curves, microscopy crucial for reliable research, particularly evaluating specific characteristics before vitro studies. study compared the patterns lines under different conditions seeding densities using trypan blue counting MTT methods. cultured heat-inactivated FBS (H.iFBS) non-heat-inactivated (Non-H.iFBS) exhibit distinct patterns, doubling times, morphologies. Cell density condition medium are two key parameters influencing metabolic activities enhancing variability. There no preference between culturing cells yet it should be considered experiment design. Thus systematic reporting essential facilitate better comparisons vivo experimental data.

Language: Английский

Citations

0
A call for standardization for secretome and extracellular vesicles in osteoarthritis: results show disease-modifying potential, but protocols are too heterogeneous—a systematic review DOI
Daniele D’Arrigo, Manuela Salerno, Luca De Marziani

et al.

Human Cell, Journal Year: 2024, Volume and Issue: 37(5), P. 1243 - 1275

Published: June 23, 2024

Language: Английский

Citations

0
The Microenvironment in an Experimental Model of Acute Pancreatitis Can Modify the Formation of the Protein Corona of sEVs, with Implications on Their Biological Function DOI Open Access
Olga Armengol-Badia, Jaxaira Maggi, Carmen Casal

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(23), P. 12969 - 12969

Published: Dec. 2, 2024

A considerable number of the physiological functions extracellular vesicles are conditioned by protein corona attached to their surface. The composition this is initially defined during intracellular synthesis, but it can be subsequently modified interactions with microenvironment. Here, we evaluated how small exposed inflammatory environment generated in acute pancreatitis and what functional changes occur as a result these modifications. Small obtained from pancreatic cell line were incubated ascitic fluid experimental rats. Using proteomic techniques, detected appearance new proteins an increase uptake certain types response induced cells. inhibition different pattern recognition receptors reversed activation, indicating that some effects could due binding damage-associated molecular patterns corona. All indicates pathologies such pancreatitis, characterized intense tissue damage, microenvironment substantially influences vesicles, thus altering behavior enhancing activity.

Language: Английский

Citations

0