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

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.

Potent induction of chondrocytic differentiation of human adipose‐derived adult stem cells by bone morphogenetic protein 6

“…The concentrations of TGF␤1, TGF␤3, and IGF-1 used for chondrogenic differentiation were based on previous studies of MSCs and ADAS cells (13)(14)(15)17,18,21,(24)(25)(26)(27)(28)30,31). To determine the dose-response effect of BMP-6, the effects of BMP-6 at a range of concentrations (5-500 ng/ml) on the expression of COL1A1, COL2A1, COL10A1, and AGC1 gene transcripts were examined, showing maximum stimulation of COL2A1 and AGC1 and suppression of COL10A1 at 500 ng/ml.…”

“…Recent studies have confirmed the existence of an abundant and easily accessible source of multipotent stem cells in subcutaneous or intraarticular adipose tissue (13)(14)(15)(16)(17)(18)(19)(20)(21). Cells obtained from adipose tissue have been termed processed lipoaspirate cells, adiposederived adult stromal/stem cells (ADAS cells), or simply adipose-derived stem cells.…”

“…Cells obtained from adipose tissue have been termed processed lipoaspirate cells, adiposederived adult stromal/stem cells (ADAS cells), or simply adipose-derived stem cells. Many studies have been undertaken to analyze and document the chondrogenic potential of ADAS cells (13)(14)(15)17,18,21,22). The induction of chondrogenesis in these cells appears to require a rounded cell shape through pellet culture or a 3-dimensional (3-D) hydrogel such as alginate, and can be augmented by a variety of chondrogenic induction agents, such as transforming growth factor ␤1 (TGF␤1), TGF␤3, dexamethasone, and ascorbic acid.…”

“…The goal of the present study was to investigate the effects of TGF␤1, TGF␤3, IGF-1, BMP-6, and dexamethasone on ADAS cell chondrogenesis; therefore, only markers associated with understanding of the process of chondrogenesis were studied. Other markers that have been associated with ADAS cells, such as adipogenic and neuronal markers (16)(17)(18)21,32), were specifically excluded in this analysis. Passage-4 ADAS cells were cultured in vitro for 7 days in a 3-D alginate culture system under various combinations of the abovementioned growth factors.…”

2010 Nicolas Andry Award: Multipotent Adult Stem Cells from Adipose Tissue for Musculoskeletal Tissue Engineering

“…To examine the multipotency of individual cells, ASCs were cultured and ring cloning was performed to select cells derived from one progenitor cell [36]. Forty-five clones were expanded through four passages and then induced for adipogenesis, osteogenesis, chondrogenesis, and neurogenesis using lineage-specific differentiation media.…”