scholarly journals The Regenerative Potential of Amniotic Fluid Stem Cell Extracellular Vesicles: Lessons Learned by Comparing Different Isolation Techniques

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Lina Antounians ◽  
Areti Tzanetakis ◽  
Ornella Pellerito ◽  
Vincenzo D. Catania ◽  
Adrienne Sulistyo ◽  
...  
2022 ◽  
Author(s):  
Kasra Khalaj ◽  
Lina Antounians ◽  
Rebeca Lopes Figueira ◽  
Martin Post ◽  
Augusto Zani

Rationale: Pulmonary hypoplasia secondary to congenital diaphragmatic hernia (CDH) is characterized by reduced branching morphogenesis, which is responsible for poor clinical outcomes. Administration of amniotic fluid stem cell extracellular vesicles (AFSC-EVs) rescues branching morphogenesis in rodent fetal models of pulmonary hypoplasia. Herein, we hypothesized that AFSC-EVs exert their regenerative potential by affecting autophagy, a process required for normal lung development. Objectives: To evaluate autophagy in hypoplastic lungs throughout gestation and establish whether AFSC-EV administration improves branching morphogenesis through autophagy-mediated mechanisms. Methods: EVs were isolated from c-kit+ AFSC conditioned medium by ultracentrifugation and characterized for size, morphology, and EV markers. Branching morphogenesis was inhibited in rat fetuses by nitrofen administration to dams and in human fetal lung explants by blocking RAC1 activity with NSC23766. Expression of autophagy activators (BECN1 and ATG5) and adaptor (SQSTM1/p62) was analyzed in vitro (rat and human fetal lung explants) and in vivo (rat fetal lungs). Mechanistic studies on rat fetal primary lung epithelial cells were conducted using inhibitors for microRNA-17 and -20a contained in the AFSC-EV cargo and known to regulate autophagy. Measurements and Main Results: Rat and human models of fetal pulmonary hypoplasia showed reduced autophagy mainly at pseudoglandular and canalicular stages. AFSC-EV administration restored autophagy in both pulmonary hypoplasia models by transferring miR-17~92 cluster members contained in the EV cargo. Conclusions: AFSC-EV treatment rescues branching morphogenesis partly by restoring autophagy through miRNA cargo transfer. This study enhances our understanding of pulmonary hypoplasia pathogenesis and creates new opportunities for fetal therapeutic intervention in CDH babies.


2017 ◽  
Vol 6 (5) ◽  
pp. 1340-1355 ◽  
Author(s):  
Carolina Balbi ◽  
Martina Piccoli ◽  
Lucio Barile ◽  
Andrea Papait ◽  
Andrea Armirotti ◽  
...  

2020 ◽  
Vol 22 (1) ◽  
pp. 38
Author(s):  
Martina Gatti ◽  
Francesca Beretti ◽  
Manuela Zavatti ◽  
Emma Bertucci ◽  
Soraia Ribeiro Luz ◽  
...  

Background—Osteoporosis is characterized by defects in both quality and quantity of bone tissue, which imply high susceptibility to fractures with limitations of autonomy. Current therapies for osteoporosis are mostly concentrated on how to inhibit bone resorption but give serious adverse effects. Therefore, more effective and safer therapies are needed that even encourage bone formation. Here we examined the effect of extracellular vesicles secreted by human amniotic fluid stem cells (AFSC) (AFSC-EV) on a model of osteoporosis in vitro. Methods—human AFSC-EV were added to the culture medium of a human pre-osteoblast cell line (HOB) induced to differentiate, and then treated with dexamethasone as osteoporosis inducer. Aspects of differentiation and viability were assessed by immunofluorescence, Western blot, mass spectrometry, and histological assays. Since steroids induce oxidative stress, the levels of reactive oxygen species and of redox related proteins were evaluated. Results—AFSC-EV were able to ameliorate the differentiation ability of HOB both in the case of pre-osteoblasts and when the differentiation process was affected by dexamethasone. Moreover, the viability was increased and parallelly apoptotic markers were reduced. The presence of EV positively modulated the redox unbalance due to dexamethasone. Conclusion—these findings demonstrated that EV from hAFSC have the ability to recover precursor cell potential and delay local bone loss in steroid-related osteoporosis.


BioFactors ◽  
2018 ◽  
Vol 44 (2) ◽  
pp. 158-167 ◽  
Author(s):  
Francesca Beretti ◽  
Manuela Zavatti ◽  
Francesca Casciaro ◽  
Giuseppina Comitini ◽  
Fabrizia Franchi ◽  
...  

2011 ◽  
Vol 20 (2) ◽  
pp. e69-e78 ◽  
Author(s):  
Dawn M. Delo ◽  
Xuan Guan ◽  
Zhan Wang ◽  
Leanne Groban ◽  
Michael Callahan ◽  
...  

2018 ◽  
pp. 21-37
Author(s):  
Sveva Bollini ◽  
Carolina Balbi ◽  
Martina Piccoli ◽  
Michela Pozzobon

Life Sciences ◽  
2015 ◽  
Vol 121 ◽  
pp. 174-183 ◽  
Author(s):  
Manuela Zavatti ◽  
Laura Bertoni ◽  
Tullia Maraldi ◽  
Elisa Resca ◽  
Francesca Beretti ◽  
...  

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