Guilak Farshid, Lott Kristen E, Awad Hani A, Cao Qiongfang, Hicok Kevin C, Fermor Beverley, Gimble Jeffrey M
Orthopaedic Research Laboratories, Department of Surgery, Duke University Medical Center, Durham, North Carolina 27710, USA.
J Cell Physiol. 2006 Jan;206(1):229-37. doi: 10.1002/jcp.20463.
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.
人脂肪来源的成体干细胞（hADAS）池在适当的体外培养条件下可呈现多种分化表型。由于脂肪组织丰富且易于获取，hADAS细胞为组织工程和其他基于细胞的治疗提供了有前景的细胞来源。然而，尚不清楚单个hADAS细胞是否能产生多种分化表型，或者每种表型是否源自异质群体中存在的特定祖细胞亚群。本研究的目的是检验单个hADAS细胞在克隆水平上具有多能性这一假设。从抽脂废弃物中分离hADAS细胞，并进行环克隆以选择源自单个祖细胞的细胞。45个克隆经过4代扩增，然后使用谱系特异性分化培养基诱导其向脂肪生成、骨生成、软骨生成和神经生成方向分化。使用组织学和生化分析确定每个谱系的定量分化标准。81%的hADAS细胞克隆分化为至少一个谱系。此外，52%的hADAS细胞克隆分化为两个或更多谱系。与脂肪细胞（12%）相比，更多克隆表达成骨细胞（48%）、软骨细胞（43%）和神经元样细胞（52%）的表型，这可能是由于反复传代后脂肪生成能力丧失所致。这些发现与hADAS细胞是一种多能成体干细胞而非仅仅是单能祖细胞混合群体的假设一致。然而，在使用功能性体内试验验证这些结果之前，谨慎解释这些结果很重要。
