title: Comparison of viability and antioxidant capacity between canine adipose-derived mesenchymal stem cells and heme oxygenase-1-overexpressed cells after freeze-thawing.
abstract: Allogenic adipose-derived mesenchymal stem cells (Ad-MSCs) are an alternative source for cytotherapy owing to their antioxidant and anti-inflammatory effects. Frozen-thawed allogenic Ad-MSCs can be used instantly for this purpose. However, the viability and function of frozen-thawed Ad-MSCs have not been clearly evaluated. The purpose of this study was to compare the viability and function of Ad-MSCs and heme oxygenase-1 (HO-1)-overexpressed Ad-MSCs in vitro after freeze-thawing. The viability, proliferation, antioxidant capacity and mRNA gene expression of growth factors were evaluated. Frozen-thawed cells showed significantly lower viability than fresh cells (77% for Ad-MSCs and 71% for HO-1 Ad-MSCs, P<0.01). However, the proliferation rate of frozen-thawed Ad-MSCs increased and did not differ from that of fresh Ad-MSCs after 3 days of culture. In contrast, the proliferation rate of HO-1-overexpressed Ad-MSCs was lower than that of Ad-MSCs. The mRNA expression levels of TGF-β, HGF and VEGF did not differ between fresh and frozen-thawed Ad-MSCs, but COX-2 and IL-6 had significantly higher mRNA expression in frozen cells than fresh cells (P<0.05). Fresh Ad-MSCs exhibited higher HO-1 mRNA expression than frozen-thawed Ad-MSCs, and fresh HO-1-overexpressed Ad-MSCs exhibited higher than fresh Ad-MSCs (P<0.05). However, there was no significant difference between fresh and frozen HO-1-overexpressed Ad-MSCs. The antioxidant capacity of HO-1-overexpressed Ad-MSCs was significantly higher than that of Ad-MSCs. Cryopreservation of Ad-MSCs negatively affects viability and antioxidant capacity, and HO-1-overexpressed Ad-MSCs might be useful to maximize the effect of Ad-MSCs for cytotherapy.
authors.affiliation: BK21 PLUS Program for Creative Veterinary Science Research, Research Institute for Veterinary Science and College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Korea.
authors.forename: Mijung
authors.initials: M
authors.lastname: Kim
authors.forename: Yongsun
authors.initials: Y
authors.lastname: Kim
authors.forename: Seunghoon
authors.initials: S
authors.lastname: Lee
authors.forename: Minyoung
authors.initials: M
authors.lastname: Kuk
authors.forename: Ah Young
authors.initials: AY
authors.lastname: Kim
authors.forename: Wanhee
authors.initials: W
authors.lastname: Kim
authors.forename: Oh-Kyeong
authors.initials: OK
authors.lastname: Kweon
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D049109:Cell Proliferation; D002470:Cell Survival; D015925:Cryopreservation / Q000662:veterinary*; D004285:Dogs; D005260:Female; D051547:Heme Oxygenase-1 / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D000975:Antioxidants; D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D051547:Heme Oxygenase-1