title: Cell culture density affects the proliferation activity of human adipose tissue stem cells.
abstract: In this study, we investigated the effect of cell density on the proliferation activity of human mesenchymal stem cells (MSCs) derived from adipose tissue (AT-MSCs) over time in culture. Passage #4 (P4) and #12 (P12) AT-MSCs from two donors were plated at a density of 200 (culture condition 1, CC1) or 5000 (culture condition 2, CC2) cells cm(-2) . After 7 days of incubation, P4 and P12 AT-MSCs cultured in CC1 were thin and spindle-shaped, whereas those cultured in CC2 had extensive cell-to-cell contacts and an expanded cell volume. In addition, P4 and P12 AT-MSCs in CC1 divided more than three times, while those in CC2 divided less than once on average. Flow cytometric analysis using 5(6)-carboxyfluorescein diacetate N-succinimidyl ester dye showed that the fluorescence intensity of AT-MSCs was lower in CC1 than in CC2. Furthermore, expression of proliferation-associated genes, such as CDC45L, CDC20A and KIF20A, in P4 AT-MSCs was higher in CC1 than in CC2, and this difference was also observed in P12 AT-MSCs. These data demonstrated that cell culture density affects the proliferation activity of MSCs, suggesting that it is feasible to design a strategy to prepare suitable MSCs using specific culture conditions.
authors.affiliation: Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
authors.forename: Dae Seong
authors.initials: DS
authors.lastname: Kim
authors.affiliation: Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
authors.forename: Myoung Woo
authors.initials: MW
authors.lastname: Lee
authors.affiliation: Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
authors.forename: Young Jong
authors.initials: YJ
authors.lastname: Ko
authors.affiliation: Top Class Plastic Surgery, Seoul, Korea.
authors.forename: Yong Hoon
authors.initials: YH
authors.lastname: Chun
authors.affiliation: Top Class Plastic Surgery, Seoul, Korea.
authors.forename: Hyung Joon
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
authors.forename: Ki Woong
authors.initials: KW
authors.lastname: Sung
authors.affiliation: Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.|Department of Health Sciences and Technology, Sungkyunkwan University, Seoul, Korea.
authors.forename: Hong Hoe
authors.initials: HH
authors.lastname: Koo
authors.affiliation: Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.|Department of Medical Device Management and Research, SAIHST, Sungkyunkwan University, Seoul, Korea.
authors.forename: Keon Hee
authors.initials: KH
authors.lastname: Yoo
mesh_terms: D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism; D002452:Cell Count; D018929:Cell Culture Techniques; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation; D005434:Flow Cytometry; D006801:Humans; D016547:Kinesins / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: C113571:CDC45 protein, human; D018797:Cell Cycle Proteins; C410089:KIF20A protein, human; D016547:Kinesins
keywords: adipose tissue; cell density; culture condition; mesenchymal stem cells; proliferation activity