Obradovic, Hristina (56444469700)Hristina (56444469700)ObradovicKrstic, Jelena (26532883400)Jelena (26532883400)KrsticTrivanovic, Drenka (54421475000)Drenka (54421475000)TrivanovicMojsilovic, Slavko (14036036900)Slavko (14036036900)MojsilovicOkic, Ivana (55749320000)Ivana (55749320000)OkicKukolj, Tamara (56001838100)Tamara (56001838100)KukoljIlic, Vesna (57190793777)Vesna (57190793777)IlicJaukovic, Aleksandra (7006010128)Aleksandra (7006010128)JaukovicTerzic, Milan (55519713300)Milan (55519713300)TerzicBugarski, Diana (35616659100)Diana (35616659100)Bugarski2025-07-022025-07-022019https://doi.org/10.1016/j.placenta.2019.05.005https://www.scopus.com/inward/record.uri?eid=2-s2.0-85065744920&doi=10.1016%2fj.placenta.2019.05.005&partnerID=40&md5=f2d236914a7b58180d00b3939496552fhttps://remedy.med.bg.ac.rs/handle/123456789/12727Introduction: Mesenchymal stem cells from Wharton's Jelly of a human umbilical cord (WJ-MSCs) are a potential tool in regenerative medicine based on their availability, proliferative potential and differentiation capacity. Since their physiological niche contains low oxygen levels, we investigated whether cultivation of WJ-MSCs at 3% O2 affects their main features. Methods: WJ-MSCs were cultured under 21% and 3% O2. Proliferation rate was followed by short and long term proliferation assays, clonogenic capacity by CFU-F assay and cell cycle and death by flow cytometry. Differentiation capacity was investigated by histochemical staining after induced differentiation. Pluripotency and differentiation markers' expression was determined by RT-PCR. Migration capacity was followed by scratch assay and mobilization from collagen, and the activity of proteolytic enzymes by zymography. Specific inhibitors of MAPK and Wnt/β-catenin pathways were used to investigate underlying molecular mechanisms. Results: Compared to standard 21% O2, cultivation of WJ-MSCs at 3% O2 did not influence their immunophenotype, while it modulated their differentiation process and enhanced their clonogenic and expansion capacity. 3% O2 induced transient change in cell cycle and prevented cell death. The expression of NANOG, OCT4A, OCT4B and SOX2 was increased at 3% O2. Both cultivation and preculturing of WJ-MSCs at 3% O2 increased their in vitro migratory capacity and enhanced the activity of proteolytic enzymes. ERK1/2 mediated WJ-MSCs’ mobilization from collagen regardless of oxygen levels, while Wnt/β-catenin pathway was activated during migration and mobilization at standard conditions. Conclusion: Culturing of WJ-MSCs under 3% O2 should be considered a credible condition when investigating their properties and potential use. © 2019 Elsevier LtdHypoxiaMesenchymal stem cells (MSC)MigrationStemnessWharton's jelly