Clonal analysis of the differentiation potential of human adipose-derived adult stem cells

Authors

  • Farshid Guilak,

    Corresponding author
    1. Departments of Surgery and Biomedical Engineering, Duke University Medical Center, Durham, North Carolina
    • Orthopaedic Research Laboratories, Department of Surgery, Division of Orthopaedic Surgery, 375 MSRB, Box 3093, Duke University Medical Center, Durham, North Carolina 27710.

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  • Kristen E. Lott,

    1. Departments of Surgery and Biomedical Engineering, Duke University Medical Center, Durham, North Carolina
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  • Hani A. Awad,

    1. Departments of Surgery and Biomedical Engineering, Duke University Medical Center, Durham, North Carolina
    Current affiliation:
    1. Department of Biomedical Engineering, Center for Musculoskeletal Research, University of Rochester, 601 Elmwood Avenue Box 639, Rochester, NY 14642.
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  • Qiongfang Cao,

    1. Departments of Surgery and Biomedical Engineering, Duke University Medical Center, Durham, North Carolina
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  • Kevin C. Hicok,

    1. Departments of Surgery and Biomedical Engineering, Duke University Medical Center, Durham, North Carolina
    Current affiliation:
    1. Macropore Biosurgery, 6740 Top Gun Street, San Diego, CA 92121.
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  • Beverley Fermor,

    1. Departments of Surgery and Biomedical Engineering, Duke University Medical Center, Durham, North Carolina
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  • Jeffrey M. Gimble

    1. Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, Louisiana
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Abstract

Pools of human adipose-derived adult stem (hADAS) cells can exhibit multiple differentiated phenotypes under appropriate in vitro culture conditions. Because adipose tissue is abundant and easily accessible, hADAS cells offer a promising source of cells for tissue engineering and other cell-based therapies. However, it is unclear whether individual hADAS cells can give rise to multiple differentiated phenotypes or whether each phenotype arises from a subset of committed progenitor cells that exists within a heterogeneous population. The goal of this study was to test the hypothesis that single hADAS are multipotent at a clonal level. hADAS cells were isolated from liposuction waste, and ring cloning was performed to select cells derived from a single progenitor cell. Forty-five clones were expanded through four passages and then induced for adipogenesis, osteogenesis, chondrogenesis, and neurogenesis using lineage-specific differentiation media. Quantitative differentiation criteria for each lineage were determined using histological and biochemical analyses. Eighty one percent of the hADAS cell clones differentiated into at least one of the lineages. In addition, 52% of the hADAS cell clones differentiated into two or more of the lineages. More clones expressed phenotypes of osteoblasts (48%), chondrocytes (43%), and neuron-like cells (52%) than of adipocytes (12%), possibly due to the loss of adipogenic ability after repeated subcultures. The findings are consistent with the hypothesis that hADAS cells are a type of multipotent adult stem cell and not solely a mixed population of unipotent progenitor cells. However, it is important to exercise caution in interpreting these results until they are validated using functional in vivo assays. © 2005 Wiley-Liss, Inc.

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