title: Fibrin matrix-supported three-dimensional organ culture of adipose tissue for selective outgrowth, expansion, and isolation of adipose-derived stem cells.
abstract: Conventional systems for isolating adipose-derived stem cells (ASC) require enzymatic digestion of adipose tissue (AT), followed by monolayer culture to the enrich the stem cell population. However, these systems are hindered by low cell yields and a lack of reproducibility. The present study was aimed at developing a unique strategy for isolating ASC based on fibrin matrix-supported three-dimensional (3-D) organ culture of native AT. Furthermore, we tried to optimize the fibrin composition by adjusting the fibrinogen and thrombin concentrations to allow rapid outgrowth and proliferation of ASC in the 3-D fibrin matrix. Human cutaneous AT fragments were encapsulated within the fibrin matrix to construct a 3-D environment and cultured under dynamic conditions. During in vitro culture the fibrin matrix provided physical support for the AT and also allowed selective outgrowth of ASC from embedded AT fragments. In situ expanded outgrown cells were recovered from the fibrin matrix by selective fibrinolysis and propagated under monolayer culture conditions. The cultured cells fulfilled the following criteria for ASC: adhesion to culture plastic, multipotent differentiation, correct immunophenotypic profile. Fibrin matrix-supported 3-D organ culture produced ASC that with high competency in terms of growth and differentiation capabilities, and resulted in a larger and more consistent cell yield than obtained with conventional culture systems. The fibrinogen and thrombin concentrations inversely affected spreading, migration, and ASC outgrowth from native AT. Our results indicate that this 3-D organ culture system for AT can be used as an efficient and reproducible method for ASC isolation.
authors.affiliation: Paik Institute for Clinical Research, Inje University, Busan 614-735, Republic of Korea. pathyang@inje.ac.kr
authors.forename: Young-Il
authors.initials: YI
authors.lastname: Yang
authors.forename: Hyeong-In
authors.initials: HI
authors.lastname: Kim
authors.forename: Jane
authors.initials: J
authors.lastname: Shelby
authors.forename: Min-Young
authors.initials: MY
authors.lastname: Choi
authors.forename: Soo-Hwa
authors.initials: SH
authors.lastname: Jang
authors.forename: Jong-Tae
authors.initials: JT
authors.lastname: Kim
authors.forename: Won-Hee
authors.initials: WH
authors.lastname: Jang
authors.forename: Chang Soo
authors.initials: CS
authors.lastname: Choi
authors.forename: Soon-Ho
authors.initials: SH
authors.lastname: Cheong
mesh_terms: D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D005337:Fibrin; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D005337:Fibrin; D005340:Fibrinogen