title: Effects Of Hypoxia in Long-Term In Vitro Expansion of Human Bone Marrow Derived Mesenchymal Stem Cells.
abstract: Mesenchymal stem cells (MSC) are considered multipotent stromal, non-hematopoietic cells with properties of self-renovation and differentiation. Optimal conditions for culture of MSC have been under investigation. The oxygen tension used for cultivation has been studied and appears to play an important role in biological behavior of mesenchymal cells. The aim is characterize MSC in hypoxia and normoxia conditions comparing their morphological and functional characteristics. Bone marrow-derived mesenchymal stem cells obtained from 15 healthy donors and cultured. MSC obtained from each donor were separated into two cultivation conditions normoxia (21% O2) and hypoxia (three donors at 1%, three donors at 2%, five donors at 3%, and four donors at 4% O2) up to second passage. MSC were evaluated for proliferation, differentiation, immunophenotyping, size and cell complexity, oxidative stress, mitochondrial activity, and autophagy. Culture conditions applied did not seem to affect immunophenotypic features and cellular plasticity. However, cells subjected to hypoxia showed smaller size and greater cellular complexity, besides lower proliferation (P < 0.002). Furthermore, cells cultured in low O2tension had lower mitochondrial activity (P < 0.03) and a reduced tendency to autophagy, although oxidative stress did not vary among groups (P < 0.39). Oxygen tension seems to be a key regulator of cellular adaptation in vitro, and metabolic effects underlying this variable remain undescribed. Heterogeneity or even lack of results on the impact of oxygen concentration used for expanding MSC highlights the need for further research, in order to optimize conditions of cultivation and expansion and achieve greater safety and therapeutic efficacy. J. Cell. Biochem. 118: 3072-3079, 2017. © 2017 Wiley Periodicals, Inc.
authors.affiliation: Celular Therapy Center of Hospital de Clinicas de Porto Alegre, Center for Experimental Research, Porto Alegre, Brazil.|Post-graduation: Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Annelise
authors.identifier: 0000-0002-4929-7525
authors.initials: A
authors.lastname: Pezzi
authors.affiliation: Celular Therapy Center of Hospital de Clinicas de Porto Alegre, Center for Experimental Research, Porto Alegre, Brazil.|Centro Universitário Ritter dos Reis, Porto Alegre, Brazil.
authors.forename: Bruna
authors.initials: B
authors.lastname: Amorin
authors.affiliation: Celular Therapy Center of Hospital de Clinicas de Porto Alegre, Center for Experimental Research, Porto Alegre, Brazil.|Post-graduation: Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Álvaro
authors.initials: Á
authors.lastname: Laureano
authors.affiliation: Celular Therapy Center of Hospital de Clinicas de Porto Alegre, Center for Experimental Research, Porto Alegre, Brazil.|Post-graduation: Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Valim
authors.affiliation: Celular Therapy Center of Hospital de Clinicas de Porto Alegre, Center for Experimental Research, Porto Alegre, Brazil.|Post-graduation: Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Alice
authors.initials: A
authors.lastname: Dahmer
authors.affiliation: Celular Therapy Center of Hospital de Clinicas de Porto Alegre, Center for Experimental Research, Porto Alegre, Brazil.|Post-graduation: Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Bruna
authors.initials: B
authors.lastname: Zambonato
authors.affiliation: Celular Therapy Center of Hospital de Clinicas de Porto Alegre, Center for Experimental Research, Porto Alegre, Brazil.|Post-graduation: Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Filipe
authors.initials: F
authors.lastname: Sehn
authors.affiliation: Celular Therapy Center of Hospital de Clinicas de Porto Alegre, Center for Experimental Research, Porto Alegre, Brazil.|Post-graduation: Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Ianaê
authors.initials: I
authors.lastname: Wilke
authors.affiliation: Feevale University, Novo Hamburgo, Brazil.
authors.forename: Lia
authors.initials: L
authors.lastname: Bruschi
authors.affiliation: Celular Therapy Center of Hospital de Clinicas de Porto Alegre, Center for Experimental Research, Porto Alegre, Brazil.
authors.forename: Maria Aparecida Lima da
authors.initials: MALD
authors.lastname: Silva
authors.affiliation: Post-graduation: Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Filippi-Chiela
authors.affiliation: Celular Therapy Center of Hospital de Clinicas de Porto Alegre, Center for Experimental Research, Porto Alegre, Brazil.|Post-graduation: Federal University of Rio Grande do Sul, Porto Alegre, Brazil.|Hematology and Bone Marrow Transplantation of Hospital de Clinicas de Porto Alegre, Porto Alegre, Brazil.
authors.forename: Lucia
authors.initials: L
authors.lastname: Silla
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D018929:Cell Culture Techniques; D015687:Cell Hypoxia; D005260:Female; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D010101:Oxygen Consumption; D013997:Time Factors
keywords: AUTOPHAGY; HYPOXIA; MESENCHYMAL STEM CELL; MITOCHONDRIAL ACTIVITY; REACTIVE OXYGEN SPECIES